Improved Technique for Identification and Enumeration of Methanogenic Bacterial Colonies on Roll Tubes by Epifluorescence Microscopy

Methanogenic fluorescent colonies can be clearly identified on roll tubes by using an epifluorescence microscope equipped with a × 2 objective. Methanogenic and nonmethanogenic colonies could be counted in roll tubes prepared from methanogenic enrichment cultures. Late-developing colonies appearing after 25 days of incubation were mainly methanogenic.     

The Effect of Incubation Temperature on the Recovery of Spores of Bacillus subtilis 8057

S ummary . The recovery of Bacillus subtilis spores was studied after different heat treatments at 95° and incubation at different temperatures in roll tubes in a gradient temperature incubator. Plate count agar and brain–heart infusion agar were used in the roll tubes. Unheated spores showed similar recoveries at 16–48° whereas heated spores had an optimum recovery temperature of c. 30.9. The rate of germination of untreated spores was greatest at c. 41° and ceased at 50°. Heated spores germinated at 52°5°, suggesting that recovery of heat-treated spores is not limited by their ability to germinate. Outgrowth of spores at different incubation temperatures was similar for germinated and ungerminated spores. Accordingly it is outgrowth rather than germination which is sensitive to temperature.     

一种嗜热厌氧纤维素降解细菌的分离纯化方法

根据纤维素降解细菌对不溶性纤维素底部的粘附作用,利用Ｈｕｎｇａｔｅ厌氧操作技术直接以不溶性纤维素粉为基质进行滚管,分离和纯化获得嗜热厌氧纤维素降解细菌。     

21. Purification of Potato Leaf Roll Virus

ABSTRACT

Potato leaf roll virus was purified by a combination of Takanami and Kubo's method and the use of the extraction and thin layer rotors and finally by centrifugation in tubes filled with beads.     

Differential carbohydrate media and anaerobic replica plating techniques in delineating carbohydrate-utilizing subgroups in rumen bacterial populations.

A basal (BC) medium devoid of added carbohydrates, a complete (CC) medium containing nine carbohydrates were developed for enumerating rumen bacteria. The colony counts on the BC medium were 85 to 100% of those obtained on the CC medium. These colonies were pinpoint size (less than or equal to mm in diameter) but increased in size (2 to 5 mm in diameter) when carbohydrates were subsequently added. With the CC medium or other media tested, the colony counts were 20 to 50% higher on plates than on roll tubes and were about 35% of the direct cell counts. The lower colony counts on roll tubes were shown to result primarily from the loss of viability due to heat stress. The DC media were found by plating techniques to be suitable for differentiating mixed rumen bacterial populations into subgroups based upon carbohydrate utilization as shown by differences in subgroup profiles found within solid and liquid fractions of rumen contents, within rumen contents from animals fed high-forage and high-grain diets, and by correct colony formations by pure cultures of rumen bacteria on appropriate DC media. With simple modifications and use of an anaerobic glove box, replica plating methods and the CC and DC media were found to be a suitable means of rapidly determining the range of utilizable carbohydrate energy sources of rumen bacteria.     

Differential carbohydrate media and anaerobic replica plating techniques in delineating carbohydrate-utilizing subgroups in rumen bacterial populations

A basal (BC) medium devoid of added carbohydrates, a complete (CC) medium containing nine carbohydrates were developed for enumerating rumen bacteria. The colony counts on the BC medium were 85 to 100% of those obtained on the CC medium. These colonies were pinpoint size (less than or equal to mm in diameter) but increased in size (2 to 5 mm in diameter) when carbohydrates were subsequently added. With the CC medium or other media tested, the colony counts were 20 to 50% higher on plates than on roll tubes and were about 35% of the direct cell counts. The lower colony counts on roll tubes were shown to result primarily from the loss of viability due to heat stress. The DC media were found by plating techniques to be suitable for differentiating mixed rumen bacterial populations into subgroups based upon carbohydrate utilization as shown by differences in subgroup profiles found within solid and liquid fractions of rumen contents, within rumen contents from animals fed high-forage and high-grain diets, and by correct colony formations by pure cultures of rumen bacteria on appropriate DC media. With simple modifications and use of an anaerobic glove box, replica plating methods and the CC and DC media were found to be a suitable means of rapidly determining the range of utilizable carbohydrate energy sources of rumen bacteria.     

A modified tube method for the cultivation and enumeration of anaerobic bacteria.

A new type of tube (the Lee tube) has been developed for use in the cultivation and enumeration of obligate anaerobes. The Lee tube is a double-walled, screw-capped tube which allows the formation of a thin cylinder of agar medium between the two walls. Anaerobiosis is achieved through deoxygenation of the deep cylinder of agar during sterilization, a minimum of head space, and use of a reducing agent to absorb oxygen introduced during the inoculation procedure. For several species of Clostridium, Bacteroides fragilis, Fusobacterium necrophorum, Veillonella alcalescens, and Pectinatus cerevisiiphilus, colony counts of cultures in the Lee tubes were comparable with those obtained in pour plates incubated in a BBL GasPak system and in anaerobic roll tubes.     

A tentative national reference procedure for isolation and enumeration of Escherichia coli from bivalve molluscan shellfish by most probable number method

In the UK several quantitative methods exist for the examination of bivalve molluscan shellfish for sewage contamination. These methods include roll tubes, pour plates and most probable number (MPN) techniques, but there is no national standard method. A comparative study was made of the most commonly used methods for detection of Escherichia coli in bivalve shellfish. Schemes employing solid media, such as the roll tube and pour plate methods, underestimated faecal contamination in shellfish tissue compared with a liquid MPN multiple test-tube method using minerals-modified-glutamate broth (MMGB) as primary enrichment medium. The composition of MMGB apparently permits repair of sublethally injured cells of E. coli. Incorporation of resuscitation stages into the pour plate technique did not yield higher counts. A standardized MPN technique for examination of bivalve molluscan shellfish for E. coli content is proposed as a possible national reference procedure pending further collaborative assessment.     

A tentative national reference procedure for isolation and enumeration of Escherichia coli from bivalve molluscan shellfish by most probable number method

In the UK several quantitative methods exist for the examination of bivalve molluscan shellfish for sewage contamination. These methods include roll tubes, pour plates and most probable number (MPN) techniques, but there is no national standard method. A comparative study was made of the most commonly used methods for detection of Escherichia coli in bivalve shellfish. Schemes employing solid media, such as the roll tube and pour plate methods, underestimated faecal contamination in shellfish tissue compared with a liquid MPN multiple test-tube method using minerals-modified-glutamate broth (MMGB) as primary enrichment medium. The composition of MMGB apparently permits repair of sublethally injured cells of E. coli. Incorporation of resuscitation stages into the pour plate technique did not yield higher counts. A standardized MPN technique for examination of bivalve molluscan shellfish for E. coli content is proposed as a possible national reference procedure pending further collaborative assessment.     

Observations and experiments on the phytoplankton of Blelham Tarn, English Lake District

Tubes composed of butyl rubber have been placed in a small English lake in an area where the water is 11-12 m deep. Each tube, when fully distended, contains over 18 000 m³ of water. The tubes are anchored to the shore and lake bottom. A boat can be rowed into them over a specially strengthened portion ofthe margin. The first was installed and its water isolated from that in the lake by the end of July 1970 and the second tube at the beginning of October 1970. The installation, maintenance, and performance of the tubes are described; as are suggested improvements. The temperature regime in the tubes closely follows that in the lake. If the chemical conditions are favourable, the seasonal cycle of Melosira in a tube is similar to that n i the lake. Contamination of the water in the tubes by that in the lake is slight and has a negligible effect on chemical conditions within them. Water can be lost from the tubes under certain conditions. The reverse does not happen. The phytoplankton communities in the tubes are similar in quality to those ofthe lake, even after 2-5 years of separation. The evidence suggests that the tubes are suitable for investigations on certain factors determining the abundance and seasonal cycles of algae in lakes and for other limnological studies.     

Influence of dietary fiber on xylanolytic and cellulolytic bacteria of adult pigs

Xylanolytic and cellulolytic bacteria were enumerated over an 86-day period from fecal samples of 10 8-month-old gilts that were fed either a control or a 40% alfalfa meal (high-fiber) diet. Fecal samples were collected from all pigs on days 0, 3, 5, 12, 25, 37, 58, and 86. Overall, the numbers of xylanolytic bacteria producing greater than 5-mm-diameter zones of clearing on 0.24% xylan roll tube medium after 24 to 36 h of incubation were 1.6 X 10(8) and 4.2 X 10(8)/g (dry weight) of feces for the control pigs and those fed the high-fiber diet, respectively. After 1 week of incubation, a large number of smaller zones of clearing (1 to 2 mm) appeared. Besides Bacteroides succinogenes and Ruminococcus flavefaciens, which produced faint zones of clearing in xylan roll tubes, three strains which closely resembled B. ruminicola hydrolyzed and used xylan for growth. The overall numbers of cellulolytic bacteria producing zones of clearing in 0.5% agar roll tube medium were 0.36 X 10(8) and 4.1 X 10(8)/g for the control pigs and those fed the high-fiber diet, respectively. B. succinogenes was the predominant cellulolytic isolate from both groups of pigs, and R. flavefaciens was found in a ratio of approximately 1 to 15 with B. succinogenes. Degradation of xylan and cellulose, measured by in vitro dry matter disappearance after inoculation with fecal samples, was significantly greater for pigs fed the high-fiber diet than that for the controls. These data suggest that the number of fibrolytic microorganisms and their activity in the large intestine of the adult pig can be increased by feeding pigs high-alfalfa-fiber diets and that these organisms are similar to those found in the rumen.     

Improved Isolation of Anaerobic Bacteria From the Gingival Crevice Area of Man

A roll tube technique (Hungate method) was employed in an attempt to cultivate a maximal portion of the organisms in the gingival crevice area of man. This technique achieves an anaerobic state by flushing the local environment with oxygen-free gas. Once collected, the crevicular debris was immediately placed into sterile oxygen-free test tubes which were flushed out by the oxygen-free gas. In this manner, the samples were weighed, dispersed, diluted, and cultured in roll tubes and plates. The medium for control (Brewer Jar technique) and Hungate techniques was Heart Infusion Agar fortified with 10% defibrinated horse blood. When the Hungate technique was used, the recovery of viable bacteria, as a percentage of the direct microscopic count, was significantly greater than plates incubated aerobically or utilizing the Brewer Anaerobic technique. Cultural counts by using the Hungate method averaged 41.3% for six samples when 90% nitrogen and 10% hydrogen were used, 70.4% for eight samples when 85% nitrogen, 10% hydrogen, and 5% carbon dioxide were used, and 63.4% for eight samples when 100% carbon dioxide was the gaseous atmosphere. At no time were cultural counts, by using anaerobic plates (Brewer Jar), more than 24% of the direct microscopic count. This suggests that exclusion of oxygen and the presence of carbon dioxide maximized recovery of gingival crevice bacteria.     

Influence of dietary fiber on xylanolytic and cellulolytic bacteria of adult pigs.

Xylanolytic and cellulolytic bacteria were enumerated over an 86-day period from fecal samples of 10 8-month-old gilts that were fed either a control or a 40% alfalfa meal (high-fiber) diet. Fecal samples were collected from all pigs on days 0, 3, 5, 12, 25, 37, 58, and 86. Overall, the numbers of xylanolytic bacteria producing greater than 5-mm-diameter zones of clearing on 0.24% xylan roll tube medium after 24 to 36 h of incubation were 1.6 X 10(8) and 4.2 X 10(8)/g (dry weight) of feces for the control pigs and those fed the high-fiber diet, respectively. After 1 week of incubation, a large number of smaller zones of clearing (1 to 2 mm) appeared. Besides Bacteroides succinogenes and Ruminococcus flavefaciens, which produced faint zones of clearing in xylan roll tubes, three strains which closely resembled B. ruminicola hydrolyzed and used xylan for growth. The overall numbers of cellulolytic bacteria producing zones of clearing in 0.5% agar roll tube medium were 0.36 X 10(8) and 4.1 X 10(8)/g for the control pigs and those fed the high-fiber diet, respectively. B. succinogenes was the predominant cellulolytic isolate from both groups of pigs, and R. flavefaciens was found in a ratio of approximately 1 to 15 with B. succinogenes. Degradation of xylan and cellulose, measured by in vitro dry matter disappearance after inoculation with fecal samples, was significantly greater for pigs fed the high-fiber diet than that for the controls. These data suggest that the number of fibrolytic microorganisms and their activity in the large intestine of the adult pig can be increased by feeding pigs high-alfalfa-fiber diets and that these organisms are similar to those found in the rumen.     

Design of a Temperature Gradient Incubator

S ummary . A temperature gradient incubator heated and cooled by circulating liquids is described which is suitable for incubating micro-organisms in liquid media and on solid media in roll tubes. The maximum deviation from a linear temperature gradient is 0·35% of the range (0·13°) even when used at temperatures up to 30° above ambient. The temperature of any set of sample wells is uniform to within 0·07° (3·5% of the difference between adjacent rows).     

Instant center of rotation estimation using the Reuleaux technique and a Lateral Extrapolation technique

A mathematical model of a rolling wheel was used to investigate the errors encountered when the Reuleaux technique is employed to estimate planar instant centers of rotation (ICRs). The investigation showed that large errors can result when this pole measurement technique is applied to objects rotating more than 12 degrees. The investigation also showed that these errors can be substantially reduced by applying a new Lateral Extrapolation technique to the pole data. When the Reuleaux technique is applied to marks on a 10cm radius wheel, the resulting offset errors from the ICR are 3.96cm for a 45 degrees roll and 1cm for a 12 degrees roll. Following lateral extrapolation, these offset errors reduce to 0.52cm for the 45 degrees roll and less than 0.04cm for the 12 degrees roll. Thus, the extrapolation technique is over seven times more accurate for a 45 degrees roll, and over 25 times more accurate for a 12 degrees roll. The extrapolation technique has been validated with the model for joints that exhibit both slip and roll, such as the knee. As joint ICR pathway measurement can be used to detect pathology, these accuracy improvements offer potential benefits for clinical applications.     

Righting and turning in mid-air using appendage inertia: reptile tails, analytical models and bio-inspired robots

Unlike the falling cat, lizards can right themselves in mid-air by a swing of their large tails in one direction causing the body to rotate in the other. Here, we developed a new three-dimensional analytical model to investigate the effectiveness of tails as inertial appendages that change body orientation. We anchored our model using the morphological parameters of the flat-tailed house gecko Hemidactylus platyurus. The degree of roll in air righting and the amount of yaw in mid-air turning directly measured in house geckos matched the model's results. Our model predicted an increase in body roll and turning as tails increase in length relative to the body. Tails that swung from a near orthogonal plane relative to the body (i.e. 0-30° from vertical) were the most effective at generating body roll, whereas tails operating at steeper angles (i.e. 45-60°) produced only half the rotation. To further test our analytical model's predictions, we built a bio-inspired robot prototype. The robot reinforced how effective attitude control can be attained with simple movements of an inertial appendage.     

Highly Stretchable,Sparse, Metallized Nanofiber Webs as Thin,Transferrable Transparent Conductors

The need for transparent conductors (TCs) that are capable of withstanding high mechanical deformation in comparison to the brittle indium tin oxide (ITO) films is paramount for roll‐to‐roll production of flexible and stretchable displays, signage systems, lighting devices and solar panels with stringent weatherability requirements. This paper reports a highly stretchable TC comprising of a web of core‐shell nanofibers, which mimics the fibrous structure of natural systems such as veins of a leaf or nerve systems. The TC web demonstrates high transparency, low sheet resistance, and unprecedented stretchability and stability over repeated stretching. The nanofiber TC web can be transferred to different substrates, which is manifested by the transfer onto an organic solar cell, demonstrating a photovoltaic performance comparable to that of a device with an ITO electrode. This work presents a technological platform, scalable for the manufacturing of large area transparent conductors for flexible and stretchable displays, electronics and solar cells on unconventional substrates such as rubber, fabric and paper.     

Silage fermentation of swine waste combined with corn

Summary Anaerobic fermentation of feedlot swine waste combined with corn was carried out in a newly designed laboratory silo. A lactic acid fermentation with rapid odor control resulted. Initial numbers of total and lactic acid bacteria (all per dry g) were 10⁷. More than 75% of the initial total population were lactobacilli that increased 27-fold at 24 h and immediately entered a phase of decreasing viable numbers. The lactobacilli were largely the streptobacterium type. Lactobacilli were counted in both plates and roll tubes, and the counts in the roll tubes were as much as 100-fold greater; the difference was attributable to anaerobic lactobacilli. After 38 d, lactobacilli were no longer found in plates, but obligately anaerobic lactobacilli persisted at 10³ through 90 d. Fecal coliform bacteria, initially present at 10⁶, were not detected after 24 h. Yeast cells, starting at 10⁶, decreased 100-fold at 3 d, and clostridia nerver exceeded 82 cells. Both groups were of minor importance in this fermentation. Virtually all the acid produced was lactic, measuring 3.83 (% dry matter) at 2 d and rising to a maximum of 12.45 at 69 d. In response, the starting pH of 6.80 decreased to 4.23 and then remained near 4 thereafter. Fumaric and succinic acid levels nerve exceeded 0.2 (% dry matter). Of the volatile fatty acids measured, acetic was maximum at 0.45 (% dry matter), but n-butyric and propionic were never more than 0.06. Fermenting a swine waste-corn mixture in a laboratory silo conferred preserved properties and diminished disease potential on a moist silage that can serve as a major component in an animal ration.The mention of firm names or trade products does not imply that they are endorsed by the U.S. Department of Agriculture over other firms or similar products not mentioned     

Microhabitat selection by the fishfly larva, Parachauliodes japonicus, in relation to its mode of respiration

SUMMARY. 1. Larvae of Parachauliodes japonicus (Megaloptera: Corydalidae) were tracked in streams using miniature radio-transmitters.
2. Larvae usually stayed under stones in shallow water for several days, but occasionally moved about on the stream bed at night.
3. Larvae possess a pair of respiratory tubes on abdominal segment VIII, which can be used as a snorkel to obtain oxygen from the atmosphere. Because respiratory rates decreased as dissolved oxygen decreased, larvae turned to air-breathing under lowered oxygen conditions. When in shallow water, they can alter respiratory mode from aquatic respiration to air breathing instantly by raising their caudal respiratory tubes to the surface of the water.
4. Microhabitat selection and respiratory behaviour are compared with those obtained previously for Protohermes grandis larvae, which belong to the same family and often inhabit the same stream as P. japonicus , but which have gill tufts on abdominal segments I-VII instead of respiratory tubes. The respiratory constraint on microhabitat preference by aquatic insects is discussed.