Enhanced mixing and mass transfer in a recirculation loop results in high cell densities in a roller bottle reactor

A recirculation loop added to a large-scale roller bottle reactor resulted in high cell densities as compared to standard roller bottles. Four different mammalian cell lines reached an average maximum density equal to 5.4 x 10(6) cells /mL (sigma = 0.263), which was between 2.13 and 2.95 times greater than the densities in roller bottles without recirculation using the same cell lines. The high densities were maintained over long durations (>25 days) while the reactor operated with continuous perfusion. The increased densities are attributed to enhanced liquid mixing and oxygen transfer that occur as a result of the recirculation loop. Models were developed that describe axial liquid flow and oxygen transfer in both the sample loop and the reactor growth chamber. Axial dispersion and oxygen transfer coefficients are presented for a variety of operating conditions. The increased oxygen transfer characteristics of the reactor allow for easy scale-up of roller bottle cultures by operating at larger volumes with greater liquid depths than conventional roller bottles permit. The surface-area-to-volume ratio in the tests performed was 0.206 versus 1.16 cm(-1) in a standard roller bottle.     

Visual wavelength discrimination by the loggerhead turtle, Caretta caretta

Marine turtles are visual animals, yet we know remarkably little about how they use this sensory capacity. In this study, our purpose was to determine whether loggerhead turtles could discriminate between objects on the basis of color. We used light-adapted hatchlings to determine the minimum intensity of blue (450 nm), green (500 nm), and yellow (580 nm) visual stimuli that evoked a positive phototaxis (the phototaxis "threshold" [pt]). Juvenile turtles were later trained to associate each color (presented at 1 log unit above that color's pt) with food, then to discriminate between two colors (the original rewarded stimulus plus one of the other colors, not rewarded) when both were presented at 1 log unit above their pt. In the crucial test, turtles were trained to choose between the rewarded and unrewarded color when the colors varied in intensity. All turtles learned that task, demonstrating color discrimination. An association between blue and food was acquired in fewer trials than between yellow and food, perhaps because some prey of juvenile loggerheads in oceanic surface waters (jellyfishes, polyps, and pelagic gastropods) are blue or violet in color.     

Fidelity and over-wintering of sea turtles

While fidelity to breeding sites is well demonstrated in marine turtles, emerging knowledge of migratory routes and key foraging sites is of limited conservation value unless levels of fidelity can be established. We tracked green (Chelonia mydas, n=10) and loggerhead (Caretta caretta, n=10) turtles during their post-nesting migration from the island of Cyprus to their foraging grounds. After intervals of 2-5 years, five of these females were recaptured at the nesting beach and tracked for a second migration. All five used highly similar migratory routes to return to the same foraging and over-wintering areas. None of the females visited other foraging habitats over the study period (units lasted on average 305 days; maximum, 1356 days), moving only to deeper waters during the winter months where they demonstrated extremely long resting dives of up to 10.2h (the longest breath-holding dive recorded for a marine vertebrate). High levels of fidelity and the relatively discrete nature of the home ranges demonstrate that protection of key migratory pathways, foraging and over-wintering sites can serve as an important tool for the future conservation of marine turtles.     

<Emphasis Type="Italic">Ilex paraguariensis</Emphasis> Pellets from a Spray-Dried Extract: Development,Characterization, and Stability

Several studies have shown the potential use of Ilex paraguariensis in developing products with the aim to protect biological systems against oxidative stress-mediated damages. In the same way, technological studies have demonstrated the feasibility of obtaining dry products, by spray-drying process, from aqueous extracts of I. paraguariensis in laboratory. The present work was designed to develop pellets by extrusion/spheronization process, from an I. paraguariensis spray-dried powder. The pellets were characterized with respect to their chemical, physical, and technological properties, and the thermal and the photostability of the main polyphenol constituents were investigated. The pellets exhibited adequate size, shape, and high process yield (78.7%), as well as a good recovery of the total polyphenols (>95%) and a good dissolution in water (89.44 to 100.05%). The polyphenols were stable against light when conditioned in amber glass bottles; unstable against heat when the samples were conditioned either in open glass bottles or in hermetically sealed glass bottles and demonstrated to be hygroscopic and sensible to the temperature, especially when stored in permeable flasks. These findings pointed to the relevance of reducing the residual moisture content of pellets as well as of conditioning them in opaque humidity tight packages under low temperatures. The feasibility of obtaining pellets from an I. paraguariensis spray-dried powder using extrusion/spheronization technique was, for the first time, demonstrated. This finding represents a novelty for the herbal products in both pharmaceutical and food fields.     

The Microflora of Hand Washed Milk Bottles

S ummary . The examination of a series of 713 milk bottles cleansed by hand washing at producer-retailer farm dairies showed that though nearly 60% attained satisfactory bacteriological standards, about 30% gave high colony counts (>600/bottle), while coli-aerogenes organisms were found in 17% and milk spoilage organisms in 25% of them. The microflora of efficiently cleansed bottles, with colony counts of <200/bottle, was dominated by micrococci and aerobic sporeforming rods. Only 3.7% of the 259 cultures from these bottles gave acid reactions in litmus milk in 72 h at 22°. Inefficiently cleansed bottles, with colony counts of >600/bottle, had quite a different type of microflora which was usually dominated by Gram negative rods (achromobacteria, nonfluorescent pseudomonads and flavobacteria). A much higher proportion (19%) of the 393 cultures from these bottles gave acid reactions in litmus milk.     

Approximation to the CDC method to determine insecticide susceptibility in leishmaniasis vectors

The Centers for Disease Control (CDC, USA) has proposed a simplified method for the determinations of insecticide resistance in adult mosquitoes, using 250 ml Wheaton bottles containing measured dosages. Insects are transferred into the bottle for 1 hour and monitored for mortality at regular intervals. In standardizing the CDC method for use with phlebotomine sand flies, effects of the solvent without insecticide were evaluated. Two colonized sand fly vector species were used: Lutzomyia longipalpis (F50 and F54) and Lutzomyia serrana (F17). Groups of 10 to 24 unfed females 1-3 days old were transferred for 1 h to Wheaton bottles with the following pretreatment: (1) without additive, (2) 0.5 ml of acetone, or (3) 1.0 ml of acetone. Three to 5 replicates were undertaken for each condition and each species. In the control bottles, the insects rested quietly and after 1 h appeared normal. In bottles with 0.5 and 1.0 ml acetone, a repellent effect was observed in L. longipalpis and L. serrana within the first 10 min. A small proportion of the L. serrana became prostrate, but recovered quickly after removal from the bottle. Field test performed with Lutzomyia quasitownsendi produced results simialar to those of the L. serrana colony flies. The insecticide bioassays were performed with L. longipalpis (F60) flies. Females were exposed to three graded doses of lambdacyhalothrin (10, 50 and 100 micrograms/bottle), and mortality was recorded at five-minute intervals. Regression lines for the 3 concentrations were compared within the context of the CDC method. The advantages of the CDC method over the WHO protocols were four: lower cost, fewer insects required, an entire group of insects exposed to the same surface, and ease of field use.     

Comparison of Macroscopic Examination, Routine Gram Stains, and Routine Subcultures in the Initial Detection of Positive Blood Cultures

Blood was cultured in two vaccum bottles containing Columbia broth with sodium polyanethol sulfonate and CO₂. Filtered air was admitted to one bottle, and the bottles were incubated at 35 C until growth was detected or for a maximum of 7 days. Bottles were examined daily for macroscopic growth. Gram stains were made routinely on the 1st, 4th, and 7th days, and samples were routinely subcultured to sheep blood agar (incubated in GasPak jar) and chocolate agar (incubated in CO₂) on the 1st and 4th days of incubation. Of 1,127 positive blood cultures, 65% were first detected by macroscopic examination, 23% were first detected by Gram stain, and 12% were first detected only by subculture.     

The Performance of the Four Anaerobic Blood Culture Bottles BacT/ALERT-FN, -FN Plus,BACTEC-Plus and -Lytic in Detection of Anaerobic Bacteria and Identification by Direct MALDI-TOF MS

Detection and identification of anaerobic bacteria in blood cultures (BC) is a well-recognized challenge in clinical microbiology. We studied 100 clinical anaerobic BC isolates to evaluate the performance of BacT/ALERT-FN, -FN Plus (BioMérieux), BACTEC-Plus and -Lytic (Becton Dickinson BioSciences) BC bottles in detection and time to detection (TTD) of anaerobic bacteria. BACTEC Lytic had higher detection rate (94/100, 94%) than BacT/ALERT FN Plus (80/100, 80%) (p<0.01) in the studied material. There was no significant difference in detection of anaerobic bacteria among the remaining bottle types. The 67 anaerobic bacteria that signalled positive in all four bottle types were analyzed to compare the time to detection (TTD) and isolates were directly identified by MALDI-TOF MS. There was a significant difference in TTD among the four bottle types (p<0.0001). The shortest median TTD was 18 h in BACTEC Lytic followed by BacT/ALERT FN (23.5 h), BACTEC Plus (27 h) and finally BacT/ALERT FN Plus (38 h) bottles. In contrast, MALDI-TOF MS performed similarly in all bottle types with accurate identification in 51/67 (76%) BacT/ALERT FN, 51/67 (76%) BacT/ALERT FN Plus, 53/67 (79%) BACTEC Plus and 50/67 (75%) BACTEC Lytic bottles. In conclusion, BACTEC Lytic bottles have significantly better detection rates and shorter TTD compared to the three other bottle types. The anaerobic BC bottles are equally suitable for direct MALDI-TOF MS for rapid and reliable identification of common anaerobic bacteria. Further clinical studies are warranted to investigate the performance of anaerobic BC bottles in detection of anaerobic bacteria and identification by direct MALDI-TOF MS.     

Effects of experimental anemia on blood ion and acid-base status of turtles during submergence in aerated water at 3 degrees C

The importance of blood hemoglobin to aquatic oxygen uptake by turtles (Chrysemys picta bellii) submerged in aerated water at 3 degrees C was tested by comparing the responses of anemic turtles (hematocrit approximately 6%) to turtles with normal hematocrits (hematocrit approximately 33%). All turtles were submerged for 42 days and blood samples were collected at 0, 7, 21, 32 and 42 days. Blood was analyzed for pH, PCO(2), PO(2), hematocrit, hemoglobin concentration ([Hb]) and plasma was analyzed for concentrations of lactate, glucose, Na(+), K(+), Ca(2+) and Mg(2+). Plasma [HCO(3)(-)] was calculated. [Hb] correlated closely with hematocrit levels. [Lactate] reached higher final values in anemic turtles (34.5+/-5.3 mmol l(-1)) than in normal turtles (14.5+/-4.6 mmol l(-1)) indicating a greater reliance of the anemic animals on anaerobic metabolism. Both groups compensated for acidosis by reduced PCO(2) and anemic turtles also had increased [Ca(2+)] and [Mg(2+)]. Blood pH fell significantly in the anemic turtles but not in the controls. Although the data indicate that the anemic turtles relied more on anaerobic metabolism than the controls, the effect was much less than expected on the basis of the reduced blood O(2) carrying capacity. Possible compensatory mechanisms utilized by the anemic turtles to minimize anaerobic metabolism are discussed.     

Similar effects of a beta-carboline and of flumazenil in negatively and positively reinforced learning tasks in mice.

Methyl beta-carboline-3-carboxylate (beta-CCM) and flumazenil (Ro15-1788) are known to be respectively an inverse agonist and an antagonist of the central benzodiazepine-receptor. Surprisingly, these two drugs have shown a similar enhancing effect in a negatively reinforced multiple-trial brightness discrimination task in mice. Thus, to evaluate the role of anxiety in this task, the action of these two drugs were compared in the same learning task with a positive or a negative reinforcement. Mice were trained for sessions of ten trials per day for six consecutive days. The sessions during the first three days took place after administration of beta-CCM (0.3 mg/kg), flumazenil (15 mg/kg) or vehicles of these drugs. A negative reinforcement (electric foot-shock) was used in a first experiment, and a positive one (food reward) in a second experiment. Results showed that, whatever the reinforcement, the two drugs enhance learning in a brightness discrimination task. The hypothesis is that flumazenil could have an inverse agonist profile in learning tasks. The question remains as to whether the flumazenil enhancing learning process results from increased arousal and/or anxiogenic factors, or from a negative modulatory influence of endogenous diazepam-like ligands for benzodiazepine receptors.     

Effects of developmental exposure to bisphenol A and ethinyl estradiol on spatial navigational learning and memory in painted turtles (Chrysemys picta)

Developmental exposure of turtles and other reptiles to endocrine disrupting chemicals (EDCs), including bisphenol A (BPA) and ethinyl estradiol (EE2, estrogen present in birth control pills), can induce partial to full gonadal sex-reversal in males. No prior studies have considered whether in ovo exposure to EDCs disrupts normal brain sexual differentiation. Yet, rodent model studies indicate early exposure to these chemicals disturbs sexually selected behavioral traits, including spatial navigational learning and memory. Thus, we sought to determine whether developmental exposure of painted turtles (Chrysemys picta) to BPA and EE2 results in sex-dependent behavioral changes. At developmental stage 17, turtles incubated at 26⁰C (male-inducing temperature) were treated with 1) BPA High (100 μg /mL), 2) BPA Low (0.01 μg/mL), 3) EE2 (0.2 μg/mL), or 4) vehicle or no vehicle control groups. Five months after hatching, turtles were tested with a spatial navigational test that included four food containers, only one of which was baited with food. Each turtle was randomly assigned one container that did not change over the trial period. Each individual was tested for 14 consecutive days. Results show developmental exposure to BPA High and EE2 improved spatial navigational learning and memory, as evidenced by increased number of times spent in the correct target zone and greater likelihood of solving the maze compared to control turtles. This study is the first to show that in addition to overriding temperature sex determination (TSD) of the male gonad, these EDCs may induce sex-dependent behavioral changes in turtles.     

THE ASSESSMENT OF THE BACTERIOLOGICAL CONDITION OF MILK BOTTLES

SUMMARY: A study of the relative values of a number of bacteriological tests for assessing the condition of milk bottles indicated that the colony count of the bottle rinse solution on yeastrel milk agar incubated for 4 days at 30°, combined with a clot-on-boiling test applied to 1 ml. of rinse in 9 ml. of sterile milk after incubation for 72 hr. at 19–20°, gave the most useful results.
The mean of the ratios of colony counts at 30° to those at 37° was 15·1, while it was as high as 22·9 for rinses with 37° of over 600 for an unsatisfactory bottle should be retained when the test is done at 30°. The thermoduric colony count of rinses of milk bottles, even when laboratory pasteurized in milk, did not provide any additional information to that given by the colony count at 30° made without pasteurization. A high proportion of the organisms in bottle rinses survived laboratory pasteurization in milk, the survival rate being highest in efficiently treated bottles.
The clot-on-boiling test gave results in general agreement with colony counts and served to indicate the potential influence of badly contaminated bottles on the keeping quality of milk placed in them. A substantial proportion of rinses with satisfactory colony counts reduced methylene blue within 48 hr. at 19–20°.
Colony counts at 37° were on the average much lower for bottles treated with steam than for bottles submitted to detergent treatment in various types of bottle washing machines. Treatment of bottles by steam or hypochlorite was more efficiently done on the farms than at the dairies.     

Feeding rhythms and diet of Farfantepenaeus paulensis under pen culture in Patos Lagoon estuary, Brazil

The feeding habits of Farfantepenaeus paulensis under pen culture in the Patos Lagoon estuary (Brazil) were studied. A total of 1074 shrimp had their stomach contents analyzed. Samples were taken bi-hourly over a 24 h period at about 10 day intervals during 2 months. F. paulensis did not exhibit strict feeding periodicity (except at D_21-22 and D_32-33). Thus, the offering of feed pellets during daylight is recommended but the rates of feeding must be further investigated. F. paulensis showed an omnivorous feeding behavior. Despite daily supply of feed pellets, natural food comprised the major part of the shrimp stomach contents. Among prey organisms, polychaetes and tanaids were the main groups recorded. Consumption of detritus and plant material decreased as shrimp grew. Intake of feed pellets increased significantly in the second month of culture (i.e. 5 g mean shrimp weight), thus it might be possible to reduce the initial input of commercial feed.     

Effect of feed type and sex on digestibility and feed efficiency utilization in black spiny‐tailed iguana (Ctenosaura pectinata)

Digestibility, feed efficiency, and the effect of sex were evaluated in black iguanas (Ctenosaura pectinata) using two commercial pellets (rabbit and chicken). The experiment was performed in 80 iguanas in a completely randomized design with a factorial arrangement 2×2 over 105 days. No differences were detected by food type in weight gain (chicken vs. rabbit: 121 vs. 154 mg/d) and daily intake (chicken vs. rabbit: 524 vs. 551 mg/d), but differences were detected (P<0.05) in feed conversion (chicken vs. rabbit: 6.45 vs. 4.47). Rabbit pellets showed higher digestibility than chicken food (P<0.01) in dry matter (59.8 vs. 41.4%) and NDF (55.4 vs. 43.6%), respectively. Sex had no effect in any of the variable responses. Black iguanas can be raised since 6 months old in captivity with commercial food designed for rabbit or broiler. No special physiological adaptations occur in black iguanas correlated with change in feeding habits during ontogeny. Zoo Biol 30:349–354, 2011. © 2010 Wiley‐Liss, Inc.     

Suitability of Aerially Sown Artificial Baits as a Technique for Poisoning Feral Goats

The preferences of a captive herd of goats with feral ancestry were evaluated for 11 artificial pest control baits and commercial stock feed pellets. A commercial stock food pellet (based on barley, bran, and oats) was the most preferred basic bait. A mixture of 2% diced Griselinia littoralis (broadleaf) leaves (a preferred natural food plant) and 2% molasses (per weight of basic bait) increased palatability of this basic bait. The best lured bait was aerially sown at I kg ha(-1) in a 380 ha area with about 50 feral goats. Only 25% of the goats were shown by the biological tracer iophenoxic acid to have eaten baits after 8 days. We conclude that one-hit aerial poisoning of goats is unlikely to be effective as a control method because most goats are unlikely to eat food from off the ground.     

乌鲁木齐市区越冬期长耳鸮的食性分析

2009～2011年间,利用食团分析法对乌鲁木齐市越冬长耳鸮(Asio otus)的食性进行分析。3年累计收集长耳鸮食团683份,辨认出1 132只猎物。分析结果表明,长耳鸮在冬季共捕食小型哺乳类6种,鸟类2种。小家鼠(Mus musculus)是最常见的食物,占总捕食量的53.45%。小型哺乳类是长耳鸮的主要食物,它在食物组成中出现的总频率为88.16%,以生物量计,小型哺乳类占食物构成的95.13%。长耳鸮的食物组成年度间差异显著,与当地猎物资源多样性和可获得性密切相关,表明长耳鸮可能采用机会主义者的捕食策略。     

完全或部分的食物剥夺对中华鳖(Pelodiscus sinensis)幼体补偿生长反应的影响:生长率的时间变化模式与体组成的变化

为了探究中华鳖（Pelodiscus sinensis）幼体的补偿生长能力,我们对中华鳖幼鳖（平均湿重9．56g）进行如下6种处理：饥饿0（对照）、1、2、3、4周,或者食物限制4周,即只投喂体湿重百分之一的食物;然后对各组进行饱食处理直到10周的实验结束为止。结果发现在饱食期的第一周各饥饿处理组的特殊生长率均显著高于对照组（P〈0．05）,但是终体重均没有赶上对照组。当饥饿或食物限制结束时,脂肪含量随着饥饿期的延长而降低,灰分和水分则表现出相反的变化趋势：脂肪含量显著低于对照（P〈0．05）,而灰分和水分则显著高于对照（P〈0．05）。蛋白含量则没有显著变化（P〉0．05）。实验结束时,除了灰分外（P〈0．05）,其他个体组成指标均恢复到对照组的水平。以上结果表明中华鳖幼体在饥饿胁迫下首先利用脂肪作为主要能源以维持生存,以及在该研究条件下完全的食物剥夺可以诱发其部分补偿生长反应．而部分食物剥夺则不能诱发此反应。     

Simple Method for Culturing Anaerobes

A simple, effective method is needed for growing obligate anaerobes in the clinical laboratory. This report describes a pre-reduced anaerobic bottle that can be taken to the bedside for direct inoculation, provides a flat agar surface for evaluation of number and morphology of colonies, and can be incubated in conventional bacteriological incubators. Each anaerobic culture set consisted of two bottles containing brain heart infusion agar and CO₂. Gentamicin sulfate (50 μg/ml) was added to one of these to inhibit facultative enteric bacilli. Comparison of the anaerobic bottles with an identical aerobic bottle which was also routinely inoculated permitted early identification of anaerobic colonies. Representative species of most anaerobic genera of proven pathogenicity for man have been isolated from this system during 10 months of routine use.     

Helicobacter canis bacteraemia in a 7-month-old child

On the basis of biochemical, phenotypic and 16S rRNA analyses, Helicobacter canis was isolated and identified from an otherwise healthy 7-month-old girl with intermittent fever. Blood cultures signalled bacterial growth after 5 days that was characterized as small gram-negative spiral rods. Subculturing on Colombia plates with 5% sheep blood, chocolate agar and brucella agar, aerobically and anaerobically as well as in a microaerophilic atmosphere, showed scanty growth after an additional 4 days. Secondarily seeded with fluid from the original bottle, the paediatric blood bottles repeatedly signalled growth after one night's incubation, whereas the conventially treated bottles did not support growth after 7 days' incubation. From the secondary seeded paediatric bottles a pure culture was isolated on chocolate agar plates, and identified as H. canis. This case indicates that blood culture systems should be compared and improved for their capacity to detect Helicobacter and related pathogenic bacteria species. Further studies are also needed to determine the importance of H. canis as a primary pathogen, and the role of cats in the possible zoonotic spread of H. canis to humans.