Characterization of the First Cultured Psychrotolerant Representative of Legionella from Antarctica Reveals Its Unique Genome Structure.

Culture-independent analysis shows that Legionella spp. inhabit a wide range of low-temperature environments, but to date, no psychrotolerant or psychrophilic strains have been reported. Here, we characterized the first cultivated psychrotolerant representative, designated strain TUM19329T, isolated from an Antarctic lake using a polyphasic approach and comparative genomic analysis. A genome-wide phylogenetic tree indicated that this strain was phylogenetically separate at the species level. Strain TUM19329T shared common physiological traits (e.g., Gram-negative, limited growth on buffered charcoal-yeast extract α-ketoglutarate [BCYEα] agar with l-cysteine requirements) with its relatives, but it also showed psychrotolerant growth properties (e.g., growth at 4°C to 25°C). Moreover, this strain altered its own cellular fatty acid composition to accumulate unsaturated fatty acid at a lower temperature, which may help maintain the cell membrane fluidity. Through comparative genomic analysis, we found that this strain possessed massive mobile genetic elements compared with other species, amounting to up to 17% of the total genes. The majority of the elements were the result of the spread of only a few insertion sequences (ISs), which were spread throughout the genome by a "copy-and-paste" mechanism. Furthermore, we found metabolic genes, such as fatty acid synthesis-related genes, acquired by horizontal gene transfer (HGT). The expansion of ISs and HGT events may play a major role in shaping the phenotype and physiology of this strain. On the basis of the features presented here, we propose a new species-Legionella antarctica sp. nov.-represented by strain TUM19329T (= GTC 22699T = NCTC 14581T). IMPORTANCE This study characterized a unique cultivated representative of the genus Legionella isolated from an Antarctic lake. This psychrotolerant strain had some common properties of known Legionella species but also displayed other characteristics, such as plasticity in fatty acid composition and an enrichment of mobile genes in the genome. These remarkable properties, as well as other factors, may contribute to cold hardiness, and this first cultivated cold-tolerant strain of the genus Legionella may serve as a model bacterium for further studies. It is worth noting that environmentally derived 16S rRNA gene phylotypes closely related to the strain characterized here have been detected from diverse environments outside Antarctica, suggesting a wide distribution of psychrotolerant Legionella bacteria. Our culture- and genome-based findings may accelerate the ongoing studies of the behavior and pathogenicity of Legionella spp., which have been monitored for many years in the context of public health.

Human change and adaptation in Antarctica: Psychological research on Antarctic wintering-over at Syowa station.

An Antarctic wintering-over station is a unique environment, as a small isolated society facing extreme survival margins. Psychological surveys have been done over ten years, including the Baum test, the Positive and Negative Affect Schedule (PANAS), the Coping Orientation to Problems Experienced (COPE), Subjective Health Complaints Inventory (SHC), the Two-Sided Personality Scale (TSPS) and medical consultations in Syowa Station, a Japanese Antarctic station to reveal the mental status of team members. Team members experienced fewer physical health risks in Antarctica than in Japan. Wintering-over team members reinterpreted situations positively and accepted their environment, sought instrumental social support, planned ahead, and used active coping skills and humour to overcome difficulties. They did not act out emotionally or deny problems. Individuals exhibited two types of coping, either stability through maintaining a previous lifestyle or flexible adjustment to a new way of life. Positive affect remained constant during the wintering-over period. In living through a harsh reality, team members drew support from the subjective feelings of an "internal relationship" with home or family in their minds. Thus, an Antarctic wintering-over station is an ideal isolated environment for psychological surveys, which can help understand future space travel and group managements in everyday societies.

Chasing Waterborne Pathogens in Antarctic Human-Made and Natural Environments, with Special Reference to Legionella spp.

Waterborne pathogenic diseases are public health issues, especially for people staying in remote environments, such as Antarctica. After repeated detection of Legionella by PCR from the shower room of Syowa Station, the Japanese Antarctic research station, we wanted to understand the occurrence of waterborne pathogens, especially Legionella, in the station and their potential sources. In this study, we analyzed water and biofilm samples collected from the water facilities of Syowa Station, as well as water samples from surrounding glacier lakes, by 16S rRNA gene-based amplicon sequencing. For Legionella spp., we further attempted to obtain a detailed community structure by using genus-specific primers. The results showed that potentially pathogenic genera were mostly localized in the station, while Legionella spp., Pseudomonas spp., and Mycobacterium spp. were also widely distributed in lakes. Genus-specific analysis of Legionella spp. within the lake environments confirmed the presence of diverse Legionella amplicon sequence variants (ASVs) that were distinctly different from the Legionella ASVs detected in the station. The majority of the Legionella ASVs inhabiting Antarctic lake habitats were phylogenetically distinct from known Legionella species, whereas the ASVs detected in the human-made station tended to contain ASVs highly similar to well-described mesophilic species with human pathogenicity. These data suggest that unexpected Legionella diversity exists in remote Antarctic cold environments and that environmental differences (e.g., temperature) in and around the station affect the community structure.IMPORTANCE We comprehensively examined the localization of potential waterborne pathogens in the Antarctic human-made and natural aquatic environment with special focus on Legionella spp. Some potential pathogenic genera were detected with low relative abundance in the natural environment, but most detections of these genera occurred in the station. Through detailed community analysis of Legionella spp., we revealed that a variety of Legionella spp. was widely distributed in the Antarctic environment and that they were phylogenetically distinct from the described species. This fact indicates that there are still diverse unknown Legionella spp. in Antarctica, and this genus encompasses a greater variety of species in low-temperature environments than is currently known. In contrast, amplicon sequence variants closely related to known Legionella spp. with reported pathogenicity were almost solely localized in the station, suggesting that human-made environments alter the Legionella community.

Evaluating Health Impact at High Altitude in Antarctica and Effectiveness of Monitoring Oxygen Saturation.

BACKGROUND: The Japanese Antarctic Research Expedition (JARE) has been conducting research activities in inland Antarctica, which is extremely cold dryland covered with a thick ice sheet. This environment may cause a health disorder called acute mountain sickness (AMS). To improve the safety of expedition members, we evaluated the impact of extreme environmental conditions on human health and the effectiveness of monitoring of hypoxia for the early detection of AMS. METHODS: In total, 9 members from JARE 59 were studied. Dome Fuji Station (Dome F), located 3,810 m above sea level (ASL), was the destination of the research party. We analyzed daily AMS scores (higher values correspond to more severe AMS-related symptoms), physiological findings, and percutaneous arterial blood oxygen saturation (SpO2) during the inland activity. We also determined the factors related to AMS scores. RESULTS: The average AMS score on arrival at Dome F was significantly higher than that at the departure point (560 m ASL). The average SpO2 level was significantly lower than that at other points. The SpO2 level correlated negatively with the AMS score in Spearman's rank correlation. Generalized estimating equations analysis showed that the AMS score was negatively associated with SpO2 level and positively associated with age. CONCLUSION: Hypoxia is a contributory factor to AMS which we can easily assess by measuring the SpO2 level with a pulse oximeter. SpO2 monitoring is a potentially useful health management tool for members in inland Antarctic expeditions. In addition, our results are helpful for understanding physiological responses and health issues in extreme environments.

Complete Genome Sequence of Novel Psychrotolerant Legionella Strain TUM19329, Isolated from Antarctic Lake Sediment.

Here, we report the complete genome sequence characteristics of Legionella strain TUM19329, a candidate for a novel psychrotolerant species isolated from Antarctic lake sediment. The genome assembly contains a single 3,750,805-bp contig with a G+C content of 39.1% and is predicted to encode 3,538 proteins.

Disease and injury statistics of Japanese Antarctic research expeditions during the wintering period: evaluation of 6837 cases in the 1st-56th parties - Antarctic health report in 1956-2016.

This study aimed to evaluate disease and injury trends among wintering members of the Japanese Antarctic Research Expedition. Obtained information is indispensable to the advancement of medical system and research. Summation was performed based on medical records of reports prepared by each expedition over the period 1956-2016. The clinical department's classification methods of the names of injuries and diseases varied among expeditions, but the names were integrated following the same classification. Of 1734 members (29 women), 6837 disease or injury cases (4 cases/person) were recorded. The rates of cases were as follows: surgical-orthopaedic (45.3%), internal medical (21.7%), dental (11.6%), dermatological (8.4%), ophthalmological (5.8%), otorhinolaryngological (5.3%), psychiatric (1.6%), and urological (0.1%) cases. There was no major change in rates by type of medical case in each expedition. This analysis made it possible to prepare medical facilities, content of physical examinations to select members, training of physicians before departure, preventive hygiene at sites, and medical research themes.

Impact of Living at the Japanese Antarctic Research Expedition Base on Urinary Status.

OBJECTIVES: Urinary disorders are generally well understood, but there are few reports on the urinary status of people living in unusual climates such as the polar regions. We studied the impact of living conditions on the urinary status of members of the Japanese Antarctic Research Expedition. METHODS: This prospective study enrolled 12 consenting members of the wintering party stationed at the Syowa Station in Antarctica between November 2012 and March 2014. The subjects completed questionnaires (the International Prostate Symptom Score [IPSS], Overactive Bladder Syndrome Score [OABSS] and Pittsburgh Sleep Quality Index [PSQI]) and kept daily voiding dairies for 3 days consecutively every 2 months. RESULTS: Compared with baseline values, the mean scores on all the questionnaires decreased during the stay in Antarctica, from 3.42 to 2.31 for the IPSS, 1.25 to 0.986 for the OABSS, and 4.58 to 3.78 on the PSQI. A significant difference was noted in the scores of seven subjects with a 3 point or more decrease in IPSS score and five members with a decrease less than 3 points (-2.22 vs 0.448, P = 0.0416). CONCLUSION: This is the first survey of the urinary status of people living in a polar environment. The findings obtained by questionnaires and voiding dairies indicated that urination and sleep status did not worsen during the stay in Antarctica.

[Anesthetic Management for Laparoscopic Surgery in a Very Elderly Patient with Severe Mitral Regurgitation].

A 90-year-old woman with severe mitral regurgitation underwent laparoscopic cholecystectomy. In her anesthetic management, we used FloTrac system and monitored arterial pressure-based cardiac index (CI), stroke volume (SV), and stroke volume variation (SVV) continuously as the indicators of intraoperative hemodynamics. Although the blood pressure fluctuated highly in Fowler position especially, we could control hemodynamics appropriately by intermittent volume infusion and adjusting administration of cardiovascular agents under monitoring these indicators. In a very elderly patient with severe mitral regurgitation, anesthesia and operation may cause severe circulatory disturbances. Therefore, it is important to select proper monitoring as stated above and maintain hemodynamics with the utmost care and attention.

Temporal changes in the skin Malassezia microbiota of members of the Japanese Antarctic Research Expedition (JARE): A case study in Antarctica as a pseudo-space environment.

The International Space Station (ISS) is located approximately 400 km above the Earth. Astronauts staying at the ISS are under microgravity and are thus unable to bathe or shower; instead, they wash their bodies using wet tissues. For astronauts, skin hygiene management is important to maintain the quality of life during long-term stays on the ISS. In Antarctica, members of a Japanese geological investigation team negotiate their way over land using snowmobiles. During their 3-month stay, they are subject to a "pseudo-space" environment similar to that experienced by ISS astronauts, including the inability to bathe or shower. In this study, temporal changes in the colonization levels of skin lipophilic fungi, Malassezia were investigated in 16 team members. Compared to the levels before their trip to Antarctica, the fold changes in Malassezia colonization levels during the researchers' stay in Antarctica were in the range of 3.0 ± 1.9 to 5.3 ± 7.5 in cheek samples, 8.9 ± 10.6 to 22.2 ± 40.0 in anterior chest samples, 6.2 ± 5.4 to 16.9 ± 25.5 in behind-the-ear samples, and 1.7 ± 0.9 to 17.4 ± 33.4 in sole-of-the-foot samples. On the scalp, the level of Malassezia colonization increased dramatically, by 96.7 ± 113.8 to 916.9 ± 1251.5 fold. During their stay in Antarctica, the team members experienced itchy scalps and produced a large number of scales. The relative proportions of Malassezia globosa and M. restricta shifted to seborrheic dermatitis/dandruff types. These results provide useful information for the development of skin hygiene management plans for astronauts staying at the ISS.

[Evaluation of perioperative blood loss and transfusion in total knee arthroplasty].

BACKGROUND: Total knee arthroplasty (TKA) is generally associated with considerable blood loss and frequent blood transfusion, which are affected by various factors. We evaluated them retrospectively. METHODS: Fifty-five patients underwent TKA at our facility between January 2008 and December 2011. We investigated retrospectively their perioperative blood loss and transfusion by using medical and anesthetic records. RESULTS: Perioperative amounts of blood loss and allogenic transfusion rates were 424 g and 9 percent (average RCC 2.4 units), respectively. All patients who had donated preoperative autologous blood were able to avoid allogenic transfusion. On the other hand, 24% of entire autologous bloods were discarded. CONCLUSIONS: There is considerable perioperative blood loss in TKA. And such factors as hemostasis, anesthetization and anticoagulation can affect it It may be possible to avoid allogenic transfusion by using preoperative autologous blood donation. However, much attention should be paid to appropriate collection of autologous bloods, because of the possibility of a high discard rate of these.

Intestinal anisakiasis treated successfully with conservative therapy: importance of clinical diagnosis.

Intestinal anisakiasis is not only a rare parasitic disease, but is also difficult to diagnose. The symptoms are not specific and are often very severe and abrupt, and the findings of clinical imaging are very remarkable. Therefore, intestinal anisakiasis is often misdiagnosed as acute abdomen or intestinal obstruction and is treated surgically. However, if intestinal anisakiasis could be diagnosed correctly, it is well treated conservatively. We experienced three cases of intestinal anisakiasis, which were diagnosed correctly and treated successfully with conservative therapy. A correct clinical history and imaging interpretation helped us diagnose intestinal anisakiasis correctly and thus treat the patients successfully with conservative therapy.

[Successful anesthetic management of laparoscopic rectopexy using rocuronium and sugammadex in a patient with Becker muscular dystrophy].

A 70-year-old man with Becker muscular dystrophy (BMD) underwent laparoscopic rectopexy under general anesthesia. For anesthetic induction, we administered total 0.6 mg · kg-1 of rocuronium with titration. Eight minutes later, train-of-four (TOF) count reached to 0 and the patient was intubated smoothly. One hundred and five minutes later, TOF ratio recovered to 100% and we administered rocuronium 10 mg additionally. Surgery was finished without any problems 95 minutes after thereafter. TOF ratio was 45% and we administered sugammadex 3 mg · kg-1, reversing neuromuscular blockade to TOF ratio 100% within 1.5 minute. The patient awoke clearly and respiratory condition was good. He was extubated without remaining neuromuscular blockade. Postoperative course was stable and there was no serious adverse effect on his muscular function intra- and post-operatively. In conclusion, rocuronium and sugammadex can be used safely and effectively in general anesthetic management for patients with muscular dystrophy. However, as the onset times and durations of these agents can be longer, we should administer these agents with titration carefully under periodic neuromuscular monitoring.

Practical experience of telehealth between an Antarctic station and Japan.

The Japanese Antarctic Research Expedition (JARE) has had a presence in Antarctica since 1956. The Syowa station is 15,000 km from Japan and evacuation of patients is impossible during the winter months. From 1956 to 2003, a total of 4932 telemedical consultations were undertaken, i.e. every member of the station needed an average of 4 medical consultations each winter. Forty five percent of the consultations were surgical or orthopaedic cases, 23% were for internal medicine and 12% were for dental problems. In the early 1980s, satellite radio-telephony was found to be useful for consultations, but did not have the ability to transmit medical pictures. Email was transmitted by the International Mobile Satellite Organization (INMARSAT) but the connection was only available every 2 hours and the maximum message size was 100 kByte. In 2004, the International Telecommunications Satellite (INTELSAT) system made a connection constantly available, and increased the maximum message size to 10 MByte. Transmission of still and moving pictures became possible. Scheduled consultations are performed monthly. A doctor in Japan can ask the patient questions and perform a proxy examination with the assistance of the Antarctic doctor. Real-time telemedicine is very effective in orthopaedic and surgical cases, skin, eye and dental troubles. Still pictures are more effective for understanding in detail skin and eye problems. Emergency consultations are effective if adequate consulting staff are available.

Use of hand-held ultrasonography to confirm the correct placement of a central venous catheter tip.

To avoid severe complications resulting from malpositioning of a central venous catheter, removal and recannulation of the catheter may be necessary, thus wasting medical equipment and increasing stress on the patient. Therefore, central venous catheters should be inserted correctly the first time. We tested whether real-time hand-held ultrasound-guided confirmation of the location of the tip of a central venous catheter inserted from the femoral vein could reduce the rate of malpositioning. Catheters were inserted using conventional methods for 65 patients, and using ultrasound guidance for 29 patients. For the latter group, when a central venous catheter was inserted, the ultrasound examiner first identified its tip located dorsal to the liver in the inferior vena cava and then fixed the catheter in position. We considered a central venous catheter to be malpositioned when its tip appeared in neither the inferior vena cava nor the right atrium-inferior vena cava junction in X-rays. Flexed or inverted catheters were also considered to be malpositioned. We compared the malpositioning rates for the ultrasound and conventional groups. Malpositioning was identified for two (6.9%) patients in the ultrasound group and 19 (29.2%) patients in the conventional group. The relative risk of ultrasound-guided versus conventional catheter insertion was 0.23 (95% confidence interval, 0.09-0.62). Our data suggest that real-time ultrasound monitoring is useful for avoiding malpositioning of central venous catheters inserted from the femoral vein.

Morbidity and health survey of wintering members in Japanese Antarctic research expedition.

STUDY DESIGN: The Japanese Antarctic Research Expedition (JARE) started in 1956. Syowa Station is the mother station of JARE at 69 degrees 00'S and 39 degrees 35'E in East Antarctica. An epidemiological survey of the wintering team of JARE was carried out based on the annual reports of JARE over the period 1956-2001. METHODS: The total number of personnel was 1,236 including 3 females. The age of personnel was from 21 to 56 (average 33.1). We examined the proportion of personnel who had contracted disease with healthy personnel, as reported by the medical department at Syowa as well as a health survey of the latest party by biochemical analyses. RESULTS: The total number of diseases was 4,760. The ratios of surgical-orthopedic, internal medical, and dental cases were 45%, 23%, and 13%, respectively. Only one death from a blizzard was recorded. In the latest party, serum levels for triglyceride, calcium, and gamma-glutamyl transpeptidase increased during the wintering period. CONCLUSION: The very low mortality of JARE may be due to the effective personnel selection and that there have been no severe accidents. There has been no deterioration in nutritional parameters in JARE recently.