Changes in egg and body temperature indicate triggering of egg desertion at a body mass threshold in fasting incubating blue petrels (Halobaena caerulea)

The triggering of transitory egg desertion in fasting and incubating blue petrels (Halobaena caerulea, nocturnal burrowing seabirds living in the subantarctic region) was investigated by continuously monitoring both body temperature (T _sto) and egg temperature (T _egg) with a telemetry system, and by measuring body mass (BM) loss. The birds were kept captive in their burrow and incubated day and night without any interruption; there was no day-night cycle in T _sto and T _egg, which averaged 39.9 °C and 32.0 °C, respectively. There was no evidence of hypothermia as a way to save energy in this fasting situation. Egg desertion occurred at night and was an abrupt and definitive phenomenon reflected by a simultaneous fall in T _egg and a peak in T _sto. After egg desertion, a distinct day-night cycle of body temperature was observed, T _sto being 0.6 °C higher during night-time (P < 0.05), probably reflecting increased nocturnal activity. BM at egg desertion averaged 166.7 ± 3.8 g in telemetered birds and 164.4 ± 1.6 g in␣a group of free-living birds. Throughout fasting, the␣specific daily BM loss remained at 46 ± 1 g · kg⁻¹ · day⁻¹, but increased sharply below a critical BM of 160.0 ± 2.5 g. Thus, fasting incubating blue petrels spontaneously desert their egg when reaching a BM threshold. This BM is very close to a critical value in fasting birds and mammals that corresponds to a critical depletion of fat stores and to a shift from lipid to protein utilization. This strongly suggests that such a metabolic shift triggers behavioural changes leading to egg desertion and refeeding, which is of great relevance to the understanding of the long-term control of food intake and BM. Accepted: 16 July 1998     

Daily torpor and energetics in a tropical mammal, the northern blossom-bat Macroglossus minimus (Megachiroptera)

Little is known about torpor in the tropics or torpor in megachiropteran species. We investigated thermoregulation, energetics and patterns of torpor in the northern blossom-bat Macroglossus minimus (16 g) to test whether physiological variables may explain why its range is limited to tropical regions. Normothermic bats showed a large variation in body temperature (T _b) (33 to 37 °C) over a wide range of ambient temperatures (T _as) and a relatively low basal metabolic rate (1.29 ml O₂ g⁻¹h⁻¹). Bats entered torpor frequently in the laboratory at T _as between 14 and 25 °C. Entry into torpor always occurred when lights were switched on in the morning, independent of T _a. MRs during torpor were reduced to about 20–40% of normothermic bats and T _bs were regulated at a minimum of 23.1 ± 1.4 °C. The duration of torpor bouts increased with decreasing T _a in non-thermoregulating bats, but generally terminated after 8 h in thermoregulating torpid bats. Both the mean minimum T _b and MR of torpid M. minimus were higher than that predicted for a 16-g daily heterotherm and the T _b was also about 5 °C higher than that of the common blossom-bat Syconycteris australis, which has a more subtropical distribution. These observations suggest that variables associated with torpor are affected by T _a and that the restriction to tropical areas in M. minimus to some extent may be due to their ability to enter only very shallow daily torpor. Accepted: 22 September 1997     

<Emphasis Type="Italic">Acinetobacter kyonggiensis</Emphasis> sp. nov., a <Emphasis Type="Italic">β</Emphasis>-glucosidase-producing bacterium,isolated from sewage treatment plant

A Gram-negative, non-motile bacterium, designated KSL5401-037^T, was isolated from a sewage treatment plant in Gwangju in the Republic of Korea and was characterized using a polyphasic taxonomic approach. Comparative 16S rRNA gene sequence analysis showed that strain KSL5401-037^T belonged to the genus Acinetobacter in the family Moraxellaceae of the Gammaproteobacteria (Brisou and Prevot, 1954). According to a 16S rRNA gene sequence analysis, it was closely related to Acinetobacter johnsonii ATCC 17909^T (97.3%), A. bouvetii 4B02^T (97.2%), and A. beijerinckii 58a^T (96.8%). Chemotaxonomic data revealed that strain KSL5401-037^T possesses an ubiquinone system with Q-8 as the predominant compound and C_16:0 (19.2%), C_18:1 ω9c (19.5%), and summed feature 3 (C_16:1 ω6c / C_16:1 ω7c, 34.1%) as the predominant cellular fatty acids. The major polar lipids detected in strain KSL5401-037^T were diphosphatidylglycerol (DPG) and, phosphatidylethanolamine (PE), followed by phosphatidylglycerol (PG) and moderate amounts of phosphatidylcholine and phosphatidylserine. The G+C content of the genomic DNA was 41.2–42.1 mol%. Strain KSL5401-037^T exhibited relatively low levels of DNA-DNA relatedness with respect to A. johnsonii DSM 6963^T (17.7%) and A. bouvetii 4B02^T (9.3%). The DNA-DNA relatedness values, biochemical, and physiological characteristics of strain KSL5401-037^T strongly support its genotypic and phenotypic differentiation from other recognized type strains of the genus Acinetobacter. Based on these data, strain KSL5401-037^T (JCM 17071^T =KEMC 5401-037^T) should be classified in the genus Acinetobacter as a type strain of novel species, for which the name Acinetobacter kyonggiensis sp. nov. is proposed.     

Daily energy expenditure of the grey mouse lemur (Microcebus murinus): a small primate that uses torpor

We aimed to investigate the pattern of utilisation of torpor and its impact on energy budgets in free-living grey mouse lemurs (Microcebus murinus), a small nocturnal primate endemic to Madagascar. We measured daily energy expenditure (DEE) and water turnover using doubly labelled water, and we used temperature-sensitive radio collars to measure skin temperature (T _sk) and home range. Our results showed that male and female mouse lemurs in the wild enter torpor spontaneously over a wide range of ambient temperatures (T _a) during the dry season, but not during the rainy season. Mouse lemurs remained torpid between 1.7–8.9 h with a daily mean of 3.4 h, and their T _sk s fell to a minimum of 18.8 °C. Mean home ranges of mouse lemurs which remained normothermic were similar in the rainy and dry season. During the dry season, the mean home range of mouse lemurs showing daily torpor was significantly smaller than that of animals remaining normothermic. The DEE of M. murinus remaining normothermic in the rainy season (122 ± 65.4 kJ day⁻¹) was about the same of that of normothermic mouse lemurs in the dry season (115.5 ± 27.3 kJ day⁻¹). During the dry season, the mean DEE of M. murinus that utilised daily torpor was 103.4 ± 32.7 kJ day⁻¹ which is not significantly different from the mean DEE of animals remaining normothermic. We found that the DEE of mouse lemurs using daily torpor was significantly correlated with the mean temperature difference between T _sk and T _a (r ²=0.37) and with torpor bout length (r ² =0.46), while none of these factors explained significant amounts of variation in the DEE of the mouse lemurs remaining normothermic. The mean water flux rate of mouse lemurs using daily torpor (13.0 ± 4.1 ml day⁻¹) was significantly lower than that of mouse lemurs remaining normothermic (19.4 ± 3.8 ml day⁻¹), suggesting the lemurs conserve water by entering torpor. Thus, this first study on the energy budget of free-ranging M. murinus demonstrates that torpor may not only reflect its impact on the daily energy demands, but involve wider adaptive implications such as water requirements. Accepted: 29 August 2000     

A real‐time PCR assay for detection and quantification of Lactococcus garvieae

Aims: To develop a rapid, sensitive, specific tool for the detection and quantification of Lactococcus garvieae in food and environmental samples. Methods and Results: A real‐time quantitative PCR (qPCR) assay with primers for CAU12F and CAU12R based on the 16S rRNA gene of L. garvieae was successfully established. The limit of detection for L. garvieae genomic DNA was 1 ng DNA in conventional PCR and 32 fg with a mean C_T value of 36·75 in qPCR. Quantification of L. garvieae vegetative cells was linear (R² = 0·99) over a 7‐log‐unit dynamic range down to ten L. garvieae cells. Conclusions: This method is highly specific, sensitive and reproducible for the detection of L. garvieae compared to gel‐based conventional PCR assays, thus providing precise quantification of L. garvieae in food and natural environments. Significance and Impact of the Study: This work provides efficient diagnostic and monitoring tools for the rapid identification of L. garvieae, an emerging pathogen in aquaculture and an occasional human pathogen from other members of the genus Lactobacillus.     

The production of xylitol from d-xylose by fermentation with Hansenula polymorpha

We have analysed the influence of the initial pH of the medium and the quantity of aeration provided during the batch fermentation of solutions of d-xylose by the yeast Hansenula polymorpha (34438 ATCC). The initial pH was altered between 3.5 and 6.5 whilst aeration varied between 0.0 and 0.3 vvm. The temperature was kept at 30 °C during all the experiments. Hansenula polymorpha is known to produce high quantities of xylitol and low quantities of ethanol. The most favourable conditions for the growth of xylitol turned out to be: an initial pH of between 4.5 and 5.5 and the aeration provided by the stirring vortex alone. Thus, at an initial pH of 5.5, the maximum specific production rate (μ_m) was 0.41 h⁻¹, the overall biomass yield (Y _x/s ^G) was 0.12 g g⁻¹, the specific d-xylose-consumption rate (q _s ) was 0.075 g g⁻¹ h⁻¹ (for t = 75 h), the specific xylitol-production rate (q _Xy ) was 0.31 g g⁻¹ h⁻¹ (for t = 30 h) and the overall yields of ethanol (Y _E/s ^G) and xylitol (Y _Xy/s ^G) were 0.017 and 0.61 g g⁻¹ respectively. Both q _s and q _Xy decreased during the course of the experiments once the exponential growth phase had finished. Received: 26 March 1998 / Received revision: 30 June 1998 / Accepted: 2 July 1998     

Aspects of body size and gonadal histology in the Antarctic toothfish, Dissostichus mawsoni, from McMurdo Sound, Antarctica

Twenty-eight specimens of the large notothenioid Dissostichus mawsoni were dissected after capture on a set line near the southern limit of its range in McMurdo Sound, Antarctica. Total length (L _T) averaged 127.3 cm (range 90–162 cm) and weight (W ) was 26.7 kg (range 10.4–60.3 kg). The length-weight relationship was W=3.44×10⁻⁵ L _T ^2.85 (n=28, r ²=0.96). Subcutaneous lipid thickness averaged 2.6 mm and showed no difference due to sex, but a significant weak relationship to W and L _T. Hepatosomatic index (I _H) was 1.6% for females and 1.7% for males; gonadosomatic index (I _G) was 0.9% for females and 0.2% for males. Although specimens were large and sexually mature, the histology indicated that the gonads of this November sample were in a resting stage. Testes lacked spermatids and spermatozoa. Oocytes were in the previtellogenic stage (91.2%) or in the first stage of vitellogenesis (8.8%). A few atretic oocytes and empty follicles indicated that some females in this sample had spawned previously. A summary of the life-cycle is also presented. Accepted: 27 September 1999     

Production of sophorolipids from whey

Sophorolipids, obtained by a two-stage process starting from deproteinized whey concentrate using Cryptococcus curvatus ATCC 20509 and Candida bombicola ATCC 22214, were compared to products from one-stage processes, using different lipidic compounds as substrates. Results showed that above all carbon source and not cultivation conditions had a distinct influence on the composition of the crude product mixture and therefore on the physicochemical and biological properties of the sophorolipids, such as, for example, surface activity, cytotoxicity and stability against hydrolases. The results were completed by corresponding data for purified mono- and diacetylated (17-hydroxyoctadecenoic)-1′,4′′-lactonized sophorolipids. Crude sophorolipid mixtures showed moderate to good surface active properties (SFT^min 39 mN m⁻¹, CMC 130 mg l⁻¹), water solubilities (2–3 g l⁻¹) and low cytotoxicities (LC₅₀ 300–700 mg l⁻¹). In contrast, purified sophorolipids were more surface active (SFT^min 36 mN m⁻¹, CMC 10 mg l⁻¹), less water soluble (max. 70 mg l⁻¹) and showed stronger cytotoxic effects (LC₅₀ 15 mg l⁻¹). Incubation of crude sophorolipid mixtures with different hydrolases demonstrated that treatment with commercially available lipases such as from Candida rugosa and Mucor miehei distinctly reduced the surface active properties of the sophorolipids, while treatment with porcine liver esterase and glycosidases had no effect. Received: 23 February 1999 / Received revision: 27 May 1999 / Accepted: 28 May 1999     

Thermogenic responses to body cooling during fever induced by Staphylococcus aureus cell walls in rabbits

Hypothalamic temperature (T _hypo) and metabolic heat production (M) were measured in seven conscious rabbits injected intravenously with either saline or with Staphylococcus aureus, (8 · 10⁷ cell walls · kg⁻¹) while being subjected to a 3-h period of ramp-like total body cooling using a chronically implanted intravascular heat exchanger. In pyrogen-injected animals cooling started (1) at the time of injection or (2) 70 min after injection. In (1) the fall in T _hypo induced by heat extraction was similar (1.0 °C) in afebrile and febrile animals. In (2) there was a transient increase in T _hypo of about 0.5 °C at a time corresponding to the start of fever resulting in a significantly smaller fall in T _hypo at the end of the 3-h cooling period (0.5 °C vs 0.9 °C, P < 0.05, n = 5). At this time in both (1) and (2) M was lower than theoretically expected from the increase in shivering threshold during fever. However, most of this effect can be explained when available data showing a decrease in thermosensitivity during S. aureus-induced fever are taken into account. After cessation of cooling in both groups of febrile animals T _hypo rose to about 1 °C higher than the precooling level, which is comparable to the fever level in a separate series of experiments with S. aureus injection without cooling (1.2 °C). Accepted: 23 September 1997     

Plasma arginine vasotocin and angiotensin II responses to body temperature elevation in the Pekin duck

The relationship between body temperature (T _b) and the plasma concentrations of arginine vasotocin (AVT) and angiotensin II (AII) was examined in conscious, adult Pekin ducks. Exposure of birds to an ambient temperature of 40 °C for 3 h increased T _b by about 1.5 °C and increased breathing rate five-fold. Plasma osmolality was elevated from the normothermic value of 294.9 ± 1.4 mosmol kg⁻¹ by about 8 mosmol kg⁻¹ Circulating AVT levels increased by about 2 pg ml⁻¹ from a basal concentration of 4.98 ± 0.15 pg ml⁻¹, a rise which could be accounted for by the change in osmotic status. Plasma AII concentrations were unchanged from the pre-heat exposure value of 31.8 ± 3.4 pg ml⁻¹. Time control birds, exposed only to an ambient temperature of 22 °C demonstrated no significant changes in any of the measured variables. The results suggest that an increased T _b has no direct effect on the circulating concentrations of AVT or AII in ducks. Accepted: 2 June 1997     

Determinants of bovine thermal response to heat and solar radiation exposures in a field environment

Continuous exposure of cattle to summer heat in the absence of shade results in significant hyperthermia and impairs growth and general health. Reliable predictors of heat strain are needed to identify this condition. A 12-day study was conducted during a moderate summer heat period using 12 Angus x Simmental (Bos taurus) steers (533 ± 12 kg average body weight) to identify animal and ambient determinations of core body temperature (T _core) and respiration rate (RR) responses to heat stress. Steers were provided standard diet and water ad libitum, and implanted intraperitoneally with telemetric transmitters to monitor T _core hourly. Visual count of flank movement at 0800 and 1500 hours was used for RR. Dataloggers recorded air temperature (T _a), and black globe temperatures (T _bg) hourly to assess radiant heat load. Analysis was across four periods and 2 consecutive days averaged within each period. Average T _a and T _bg increased progressively from 21.7 to 30.3°C and 25.3 to 34.0°C, respectively, from the first to fourth periods. A model utilizing a quadratic function of T _a explained the most variation in T _core (R ² = 0.56). A delay in response from 1 to 3 h did not significantly improve R ² for this relationship. Measurements at 0800 and 1500 hours alone are sufficient to predict heat strain. Daily minimum core body temperature and initial 2-h rise in T _a were predictors of maximum core temperature and RR. Further studies using continuous monitoring are needed to expand prediction of heat stress impact under different conditions.     

Anomalies in the growth kinetics of Saccharomyces cerevisiae strains in aerobic chemostat cultures

Aerobic glucose-limited chemostat cultivations were conducted with Saccharomyces cerevisiae strains NRRL Y132, ATCC 4126 and CBS 8066, using a complex medium. At low dilution rates all three strains utilised glucose oxidatively with high biomass yield coefficients, no ethanol production and very low steady-state residual glucose concentrations in the culture. Above a threshold dilution rate, respiro-fermentative (oxido-reductive) metabolism commenced, with simultaneous respiration and fermentation occurring, which is typical of Crabtree-positive yeasts. However, at high dilution rates the three strains responded differently. At high dilution rates S. cerevisiae CBS 8066 produced 7–8 g ethanol L⁻¹ from 20 g glucose L⁻¹ with concomitant low levels of residual glucose, which increased markedly only close to the wash-out dilution rate. By contrast, in the respiro-fermentative region both S. cerevisiae ATCC 4126 and NRRL Y132 produced much lower levels of ethanol (3–4 g L⁻¹) than S. cerevisiae CBS 8066, concomitant with very high residual sugar concentrations, which was a significant deviation from Monod kinetics and appeared to be associated either with high growth rates or with a fermentative (or respiro-fermentative) metabolism. Supplementation of the cultures with inorganic or organic nutrients failed to improve ethanol production or glucose assimilation. Journal of Industrial Microbiology & Biotechnology (2000) 24, 231–236. Received 09 August 1999/ Accepted in revised form 18 December 1999     

Comparison of exopolysaccharide production by strains of Lactobacillus rhamnosus and Lactobacillus paracasei grown in chemically defined medium and milk

Exopolysaccharide (EPS) production was compared among three strains of lactobacilli. Lactobacillus rhamnosus strain 9595M can be classified among the highest EPS-producing strains of lactic acid bacteria reported to date with a maximum EPS production of 1275 mg L⁻¹. Under controlled pH, no significant differences in the quantity of EPS produced could be detected between carbon source (glucose or lactose) or fermentation temperature (32 or 37°C). In milk, strains ATCC 9595M and R produced more than 280 mg L⁻¹ EPS whereas strain Type V produced less than 80 mg L⁻¹ EPS. Journal of Industrial Microbiology & Biotechnology (2000) 24, 251–255. Received 10 September 1999/ Accepted in revised form 22 December 1999     

Screening and characterization of bioflocculant produced by isolated Klebsiella sp.

Sixteen strains of polymer-producing bacteria were isolated from the activated sludge samples taken from two seafood processing plants in Southern Thailand. Their culture broths possessed the ability to flocculate kaolin suspension in the presence of 1% CaCl₂. Based on the flocculating activity, the strain S11 was selected and identified to be a Klebsiella sp. using the partial 16S rRNA sequencing method. The growth of the isolated Klebsiella sp. was maximal (1.026 g l⁻¹ dry cell mass) after 1 day cultivation while the highest polymer yield (0.973 g l⁻¹) was achieved after 5 days cultivation. The flocculating activity of the culture broth, however, was highest after 2 days cultivation. The polymer was identified to be an acidic polysaccharide containing neutral sugar and uronic acid as its major and minor components, respectively. Results on the properties of the partially purified polysaccharide from Klebsiella sp. S11 revealed that it consisted of galactose, glucose and mannose in an approximate ratio of 5:2:1. It was soluble in acidic or basic solutions but not in organic solvents. Its molecular mass was greater than 2 × 10⁶ Da. Infrared spectra showed the presence of hydroxyl, carboxyl and methoxyl groups in its molecules. Differential scanning calorimetry of the polysaccharide indicated the crystalline melting point (T _m) at 314 °C. The optimum dosage of polysaccharide to give the highest flocculating activity was 15 mg l⁻¹ in the presence of 1% CaCl₂. Received: 8 February 1999 / Received last revision: 4 June 1999 / Accepted: 4 June 1999     

<Emphasis Type="Italic">Oceanobacillus manasiensis</Emphasis> sp. nov., a moderately halophilic bacterium isolated from the salt lakes of Xinjiang,China

Three Gram reaction positive, rod-shaped, moderately motile halophilic bacterial strains, designated YD3-56^T, YD16, and YH29, were isolated from the sediments of Manasi and Aiding salt lakes in the Xinjiang region of China, respectively. The strains grew optimally at 30–37°C, pH 8–11, in the presence of 5–10% (w/v) NaCl. Phylogenetic analysis based on 16S rRNA gene sequences revealed that the strains were closely related to members of the genus Oceanobacillus, exhibiting 99.1–99.2% similarity to O. kapialis KCTC 13177^T, 99.2–99.3% to O. picturae KCTC 3821^T, and 94.2–96% sequence similarity to other described Oceanobacillus species. SDS-PAGE of whole cell proteins preparations demonstrated that the strains exhibited high similarity to each other, but distinguished from O. kapialis KCTC 13177^T and O. picturae KCTC 3821^T (75%). DNA-DNA hybridization revealed that the similarity between the representative strain YD3-56^T and O. kapialis KCTC 13177^T was 35.3%, and the similarity between YD3-56^T and O. picturae KCTC 3821^T was 22.3%. Chemotaxonomic analysis of the strains showed menaquinone-7 was the predominant respiratory quinine. Major cellular fatty acids were anteiso-C_15:0 and anteiso-C_17:0. The polar lipid pattern for strain YD3-56^T predominantly contained phosphatidylcholine, and trace to moderate amounts of phosphatidyl ethanolamine and hydroxy-phosphatidyl ethanolamine. The diamino acid in murein was meso-diaminopimelic acid. The DNA G+C content of the strains was 39.7–40.1 mol%. On the basis of these results, the three strains should be classified as a novel species of the genus Oceanobacillus, for which the name Oceanobacillus manasiensis sp. nov. has been proposed, with the type strain as YD3-56^T (=CGMCC 1.9105^T =NBRC 105903^T).     

The influence of cold stress and a 36- h fast on the physiological responses to prolonged intermittent walking in man

In a previous study, rectal temperature (T _re) was found to be lower, and oxygen consumption (V˙O₂) and the respiratory exchange ratio (R) were higher in a cold (+5°C), wet and windy environment (COLD), compared with a thermoneutral environment during intermittent walking at ≈30% of peak V˙O₂ (Weller AS, Millard CE, Stroud MA et al. Am J Physiol 272:R226–R233, 1997). The aim of the present study was to establish whether these cold-induced responses are influenced by prior fasting, as impaired thermoregulation has been demonstrated in cold-exposed, resting men following a 48-h fast. To address this question, eight men attempted a 360-min intermittent (15 min rest, 45 min exercise) walking protocol under COLD conditions on two occasions. In one condition, the subjects started the exercise protocol ≈120 min after a standard meal (FED/COLD), whereas in the other the subjects had fasted for 36 h (FASTED/COLD). The first two exercise periods were conducted at a higher intensity (HIGHER, 6 km · h⁻¹ and 10% incline), than the four subsequent exercise periods (LOW, 5 km · h⁻¹ and 0% incline). There was no difference in the time endured in FED/COLD and FASTED/COLD. In FASTED/COLD com pared with FED/COLD, R was lower during HIGHER and LOW, and T _re was lower during LOW, whereas there was no difference in V˙O₂, mean skin temperature and heart rate. Therefore, although the 36-h fast impaired temperature regulation during intermittent low-intensity exercise in the cold, wet and windy environment, it was unlikely to have been the principal factor limiting exercise performance under these experimental conditions. Accepted: 26 August 1997     

Body temperatures in free-flying pigeons

We examined the relationship between body temperature (T_b) of free flying pigeons and ambient water vapor pressure and temperature. Core or near core T_b of pigeons were measured using thermistors inserted into the cloaca and connected to small transmitters mounted on the tail feathers of free flying tippler pigeons (Columba livia). Wet and dry bulb temperatures were measured using modified transmitters mounted onto free-flying pigeons. These allowed calculation of relative humidity and hence water vapor pressure at flight altitudes. Mean T_b during flight was 42.0 ± 1.3 °C (n = 16). Paired comparisons of a subset of this data indicated that average in-flight T_b increased significantly by 1.2 ± 0.7 °C (n = 7) over that of birds at rest (t = −4.22, P < 0.05, n = 7) within the first 15 min of takeoff. In addition, there was a small but significant increase in T_b with increasing ambient air (T_a) when individuals on replicate flights (n = 35) were considered. Inclusion of water vapor pressure into the regression model did not improve the correlation between body temperature and ambient conditions. Flight T_b also increased a small (0.5 °C) but significant amount (t = 2.827, P < 0.05, n = 8) from the beginning to the end of a flight. The small response of T_b to changing flight conditions presumably reflects the efficiency of convection as a heat loss mechanism during sustained regular flight. The increase in T_b on landing that occurred in some birds was a probable consequence of a sudden reduction in convective heat loss. Accepted: 2 February 1999     

Thermal status of wet-suited divers using closed circuit O2 apparatus in sea water of 17–18.5°C

A wet suit may not provide adequate thermal protection when diving in moderately cold water (17–18°C), and any resultant mild hypothermia may impair performance during prolonged diving. We studied heat exchange during a dive to a depth of 5 m in sea water (17–18.5°C) in divers wearing a full wet suit and using closed-circuit oxygen breathing apparatus. Eight fin swimmers dived for 3.1 h and six underwater scooter (UWS) divers propelled themselves through the water for 3.7 h. The measurements taken throughout the dive were the oxygen pressure in the cylinder and skin and rectal temperatures (T _re). Each subject also completed a cold score questionnaire. The T _re decreased continuously in all subjects. Oxygen consumption in the fin divers (1.40 l · min⁻¹) was higher than that of the UWS divers (1.05 l · min⁻¹). The mean total insulation was 0.087°C · m² · W⁻¹ in both groups. Mean body insulation was 37% of the total insulation (suit insulation was 63%). The reduction in T _re over the 1st hour was related to subcutaneous fat thickness. There was a correlation between cold score and T _re at the end of 1 h, but not after that. A full wet suit does not appear to provide adequate thermal protection when diving in moderately cold water. Accepted: 21 January 1997     

Construction of a food-grade multiple-copy integration system for Lactococcus lactis

A food-grade vector system was developed that allows stable integration of multiple plasmid copies in the chromosome of Lactococcus lactis. The vector consists of the plus origin of replication (Ori⁺) of the lactococcal plasmid pWV01, the sucrose genes of the lactic acid bacterium Pediococcus pentosaceus PPE1.0 as selectable marker, a multiple-cloning site, and a lactococcal DNA fragment of a well-characterized chromosomal region. The system includes two L. lactis strains, LL108 and LL302, which produce the pWV01 RepA protein essential for replication of the Ori⁺ vectors. These helper strains allow the construction and isolation of the replicating form of the integration plasmids from a homologous background. Single-cross-over integration of the plasmids in L. lactis MG1363 resulted in amplifications to a level of approximately 20 copies/chromosome after selection of the transformants on medium containing sucrose as the only fermentable sugar. The amplifications were stable under selective growth conditions. In glucose-containing medium a limited loss of integrated plasmid copies was detected at a rate of (7.5–15) × 10⁻² copies per generation. One strain, MG124, was isolated that had retained 11 integrated copies after a period of 120 generations of non-selective growth. These results show that the single-cross-over integration system described here represents a simple procedure for the engineering of stable food-grade strains carrying multiple copies of a gene of interest. Received: 23 September 1997 / Received revision: 21 November 1997 / Accepted: 21 November 1997     

MR Proton Relaxivities of Some Ions, Albumin, and Cholesterol Added to Odontogenic Jaw Cysts

The relaxation rates (1 / T ₁ and 1 / T ₂) in cysts have already been analyzed in terms of materials such as albumin, cholesterol, manganese, iron, and copper. However, the relaxivities of these materials have not been determined yet. In this work, five sets containing the ions, albumin, and cholesterol were prepared by addition of increasing concentration of one material to each set. The relaxation times in these sets were measured by MRI, and the relaxation rates were fitted versus concentrations. The slopes of the fits were used as relaxivities. The (r ₁, r ₂) values of manganese, iron, and copper in mM⁻¹ s⁻¹, and those of albumin and cholesterol in (g/dl)⁻¹ s⁻¹ were found to be (32.64, 89.77), (0.31, 1.19), (0.5, 1.479), (0.01, 0.066) and (0.03, 0.458), respectively. The r ₂/r ₁ ratio ranged from 2.75 to 15.27. Manganese is an efficient relaxer, but iron and copper are poor ones. Albumin and cholesterol are efficient relaxers for only T ₂. The contribution of water associated with native manganese of the cystic fluid to T ₁ was 0.268 s⁻¹, whereas those of water associated with native manganese, albumin, cholesterol, and iron to T ₂ were 0.736, 0.185, 0.092, and 0.076 s⁻¹, respectively. The other contributions were much smaller than 0.076 s⁻¹. Manganese is most likely the compound altering T ₁-weighted images between different jaw cysts, whereas manganese and albumin are most likely the compounds altering the T ₂-weighted images. Present data suggest that such alterations may be used to separate jaw cysts from other jaw masses. The high r ₂/r ₁ suggests that T ₂ is a more convenient parameter than T ₁ for diagnostic use. This work is a part of the PhD thesis of U. Nezih Yilmaz supervised by R. Guner.