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Notes: Scanning electron microscopy revealed spores of Nosema apis and Thelohania fibrata to be egg-shaped, but only the mature spore of T. fibrata was shown to possess a horseshoe-like concavity at the posterior pole. Freezeetched preparations indicated that this concavity was due to a thin area of the spore coat. Freeze-etching studies also show spores of N. apis do possess an umbrella-shaped polaroplast, and a polar filament which is arranged in a double layer with over 30 coils. The spore of T. fibrata contains a pear-shaped arrangement of the polaroplast membrane, and a polar filament arranged in a single layer of 22 coils.

Notes: The fiber constituents and connections of the calyces  -  the input-receiving regions  -  of the corpora pedunculata (“mushroom bodies”) were studied in reduced silver preparations from the American cockroach, Periplaneta americana (L.). In the outer synaptic layer of the calyces five fiber classes were distinguished, the first three of which arise outside the mushroom body. (1) Four highly similar neurons with somata near the optic lobe branch into different parts of the ipsiateral protocerebrum, including both calyces. Their fibers are highly constant in arrangement and position and contain small nucleus-like bodies. (2) The tractus olfactorio-globularis (sensu lato) emits fiber groups which course along the calycal walls as “calycal tracts” before ultimately dissipating into the synaptic layer. Variability within these tracts is described. (3) Fibers of undertermined origin outside the mushroom body radiate from the calycal center outwards through the synaptic layer. (4) From the inner calycal layer of neurites belonging to intrinsic mushroom-body neurons, perpendicular collaterals enter the synaptic layer. (5) Intrinsic-neuron somata near the calycal rim emit fibers which course tangentially within the synaptic layer from calycal rim to center. These fibers form a special peripheral zone in the pedunculus.The predominant presumably afferent calycal fiber class is that derived from the tractus olfactorio-globularis. No evidence was found for tracts from optic lobe to calyces. On this basis, and in light of the experimental and comparative anatomical literature, it is suggested that the corpora pedunculata of P. americana and other pterygotes are fundamentally second-order antennal sensory processing centers.Conflicting observations in earlier reports are critically discussed.

Notes: The epidermis of Eisenia is covered by a cuticle and rests on a basement lamella. The cuticle, which is resistant to a variety of enzymes, is composed of non-striated, bundles of probable collagen fibers that are orthogonally oriented and are embedded in a proteoglycan matrix. The basement lamella consists of striated collagen fibers with a 560 Å major periodicity. Proximity and morphology suggest that the epidermis may contribute to both the cuticle and the basement lamella  -  that is, the single tissue may synthesize at least two types of collagen. The epidermis is a pseudostratified epithelium containing three major cell types (columnar, basal and gland) and a rare fourth type with apical cilia. The esophagus is lined by a simple cuticulated epithelium composed predominantly of a single cell type, which resembles the epidermal columnar cell. Rare gland cells occur in the esophageal epithelium, but basal cells are lacking.

Notes: Gill development begins on the sixth day of incubation at 10°C and is complete by 31 days (hatching). Gill arches are formed by fusion and perforation of ectoderm and endoderm across the pharyngeal wall. A primary branchial artery forms within each arch and a second branchial artery forms as a branch from its ventral end. A series of filament loop vessels forms connecting the two arteries and when several are patent a unidirectional blood flow is established via afferent (second) branchial artery, filament loop vessels to efferent (primary) branchial artery. Part of the efferent branchial artery just above its junction with the afferent branchial artery constricts and occludes. It is suggested that this change in the pattern of blood flow is dependent on differences in resistance of the two branchial arteries. A later extension of the gill ventrally is thought not to be homologous with similar regions in elasmobranchs and Acipenser.

Notes: Scanning and transmission electron microscopy studies were conducted on the antennal sensory sensilla of the hymenopteran parasitoid, Cardiochiles nigriceps Viereck, of the family Braconidae. Distinct morphological differences were found between the chemoreceptors of the male and female. Curved, non-fluted, thin-walled sensilla were found to be very abundant on the male and restricted in location and number on the female. Trichoid, placoid and fluted basiconic sensilla were numerous on the antennal flagella of both sexes. Smooth basiconic sensilla were restricted in number to one per flagellar segment in both sexes. Behavioral data suggest that bent-tipped, thick-walled sensilla unique to the female are involved in detecting a chemical(s) emitted from the host, Heliothis virescens (Fab.).

Notes: The biomechanical role of the mammalian clavicle and the functional significance of the aclaviculate condition were investigated. Shoulder movements in rats (Rattus norvegicus) with excised clavicles were compared to those of normal rats by biplanar plate radiography. Shoulder movements during walking of the claviculate American opossum (Didelphis marsupialis), and aclaviculate raccoons (Procyon lotor) and cats (Felis domestica) were compared by biplanar cineradiography.The mammalian clavicle, where present in its complete form, exerts both a “spoke” and a “strut” effect on shoulder movement. By maintaining a fixed distance between the acromion and manubrium, the clavicle ensures that relative movement between these structures is arcuate. Aclaviculate mammals, in contrast, have linear shoulder excursions that are nearly parallel or slightly oblique to the median plane, depending on the conformation of the thorax. Medial collapse of the shoulder in aclaviculate rats demonstrates that the clavicle is under compression, and thus acts as a strut.Reduction or loss of the clavicle, which has occurred independently in numerous mammalian phylogenies, has been regarded as an adaptation for greater shoulder movement and hence increased stride. However, on present evidence clavicular reduction in cursorial mammals appears to be more directly related to a linear excursion of the shoulder joint and a restriction of limb movements to a sagittal plane.

Notes: The tail of Teratoscincus scincus has dorsal scales that have tubercles on their dorsal and ventral surfaces. Sounds are produced when these rub past each other as the excited animal moves its tail. The relative movement of scales is intensified by caudal torsion. The frequencies of the sounds cover a range from 9 to 25 kops and thus, differ from those produced during vocalizations.

Notes: The locomotor function of the caudal muscle cells of ascidian larvae is identical with that of lower vertebrate somatic striated (skeletal) muscle fibers, but other features, including the presence of transverse myomuscular junctions, an active Golgi apparatus, a single nucleus, and partial innervation, are characteristic of vertebrate myocardial cells.Seven stages in the development of the compound ascidian Distaplia occidentalis were selected for an ultrastructural study of caudal myogenesis. A timetable of development and differentiation was obtained from cultures of isolated embryos in vitro.The myoblasts of the neurulating embryo are yolky, undifferentiated cells. They are arranged in two bands between the epidermis and the notochord in the caudal rudiment and are actively engaged in mitosis.Myoblasts of the caudate embryo continue to divide and rearrange themselves into longitudinal rows so that each cell simultaneously adjoins the epidermis and the notochord. The formation of secretory granules by the Golgi apparatus coincides with the onset of proteid-yolk degradation and the accumulation of glycogen in the ground cytoplasm.Randomly oriented networks of thick and thin myofilaments appear in the peripheral sarcoplasm of the muscle cells of the comma embryo. Bridges interconnect the thick and thin myofilaments (actomyosin bridges) and the thick myofilaments (H-bridges), but no banding patterns are evident. The sarcoplasmic reticulum (SR), derived from evaginations of the nuclear envelope, forms intimate associations (peripheral couplings) with the sarcolemma.Precursory Z-lines are interposed between the networks of myofilaments in the vesiculate embryo, and the nascent myofibrils become predominantly oriented parallel to the long axis of the muscle cell.Muscle cells of the papillate embryo contain a single row of cortical myofibrils. Myofibrils, already spanning the length of the cell, grow only in diameter by the apposition of myofilaments. The formation of transverse myomuscular junctions begins at this stage, but the differentiating junctions are frequently oriented obliquely rather than orthogonally to the primary axes of the myofibrils.With the appearance of H-bands and M-lines, a single perforated sheet of sarcoplasmic reticulum is found centered on the Z-line and embracing the I-band. The sheet of SR establishes peripheral couplings with the sarcolemma.In the prehatching tadpole, a second collar of SR, centered on the M-line and extending laterally to the boundaries with the A-bands, is formed. A single perforated sheet surrounds the myofibril but is discontinuous at the side of the myofibril most distant from the sarcolemma. To produce the intricate architecture of the fully differentiated collar in the swimming tadpole (J. Morph., 138: 349, 1972). the free ends of the sheet must elevate from the surface of the myofibril, recurve, and extend peripherally toward the sarcolemma to establish peripheral couplings.Morphological changes in the nucleus, nucleolus, mitochondria, and Golgi bodies are described, as well as changes in the ground cytoplasmic content of yolk, glycogen, and ribosomes.The volume of the differentiating cells, calculated from the mean cellular dimensions, and analyses of cellular shape are presented, along with schematic diagrams of cells in each stage of caudal myogenesis. In an attempt to quantify the differences observed ultrastructurally, calculations of the cytoplasmic volume occupied by the mqjor classes of organelles are included.Comparison is made with published accounts on differentiating vertebrate somatic striated and cardiac muscles.

Notes: Anolis embryos have limb buds at the time eggs are laid and require about 39 days to complete development at 28°C. Rathke's pouch is present at five days, and the subdivisions of the adenohypophysis are differentiated by ten days after oviposition. The cells of the rostral half of the pars distalis (PD) are derived from the anterior face of Rathke's pouch; cells of the caudal half from the posterior face. Lateral lobe cells differentiate on the lateral margins of the developing caudal PD, and knob-like outgrowths of this tissue attach to the walls of the diencephalon to form the pars tuberalis (PT). Subsequently, the cells of the PT lose their connection with the PD and become a pair of flattened oblong plaques. They reach maximal size in midincubation, and are gradually invaded by nervous elements and incorporated into the walls of the hypothalamus. Electron micrographs demonstrate that the embryonic PT is secretory.Ultrastructurally the pars intermedia (PI) and PD are composed of parenchymous secretory cells in a framework of stellate cells. Stellate cells surround the lumen of Rathke's pouch and are connected laterally by complex junctions that exclude the secretory cells from the luminal surface. They extend in sheet-like processes among the secretory cells to the outer margin of the gland where they form a partial sheath within the basal lamina around the secretory tissue. As development proceeds, the lumen becomes subdivided and the resulting reduced lumina are recognizable as the forerunners of the follicles of the adult adenohypophysis.The cells of the PI are differentiated into secretory or stellate cells halfway through incubation. At this time only half of the cells of the PD can be so classified. Four of the five granulated cell types described in the adult are recognizable by mid-incubation; the fifth cell type (prolactin cell) becomes distinguishable within ten days thereafter, and at hatching appears to be actively synthesizing secretory products.

Notes: The hydranth of the gymnoblastic hydroid Syncoryne tenella is invested by a cuticle approximately 530 mμ thick which is continuous with the periderm of the hydrocaulus. The ectodermal cells of the hydranth possess regularly spaced microvilli orientated with their long axis perpendicular to the ectodermal surface. The microvilli project into the cuticle, and probably serve to anchor the cuticle to the ectoderm. In the hydrocaulus the periderm is loosely applied to the ectoderm: in this region microvilli are absent from ectodermal cells. The periderm is a layered structure composed of finely filamentous material. No structural basis is found for the previously reported differential staining of peridermal layers in the hydrocaulus.

Notes: The nuchal organs of polychaetes from four different families (Nereidae, Nephtyidae, Phyllodocidae and Glyceridae) were examined with the light and electron microscopes. In each case, the organ consists of ciliated cells and primary sensory elements. The ciliated cells are similar to the cells of the adjacent epidermis but bear motile cilia. Primary sensory neurons are situated within the organs in Nephtyidae and Phyllodocidae, but are located within the brain in Nereidae and Glyceridae. Each sensory cell gives rise to a distal process which penetrates between the ciliated cells to form an apical sensory bulb bearing modified cilia. Apically these processes are lined with juxtamembranous plaques. The ciliated cells are innervated by efferent axons from the brain, and in Nereis the axons appear to be peptidergic. The elements comprising the nuchal organs closely resemble those of the vertebrate olfactory mucosa.

Notes: The cuticle of five species of Oligochaeta, chosen to represent differences in size and a variety of biotopes, was studied electron microscopically after fixation with the acrolein-TAPO-osmium tetroxide method. Five distinct layers in the cuticle of all studied species were found. Staining with lead and uranyl ions or with silver proteinate visualized basically the same structural components of the cuticle, but the degree of electron opacity and the distribution of the electron-opaque stain in these components differed according to the staining method used. Since the acrolein-TAPO-osmium tetroxide method visualized the cuticular zones preferentially stained by Thiéry's silver proteinate method, it was concluded that the TAPO method may be considered suitable for the visualization of polysaccharides. Staining with phosphotungstic acid provided some information on the composition of the cuticle of Oligochaeta not obtained by staining ultrathin sections with lead and uranyl ions nor with silver proteinate. The conclusion is that phosphotungstic acid binds to polysaccharides which do not contain vicglycol groups nor active sites responsible for the positive reaction with lead and uranyl salts. Structural components in the cuticle of the oligochaetes studied were characteristic for each species. The taxonomic value of such components, however, must be confirmed by examination of a larger number of species of oligochaetes.

Notes: Laboratory-reared outgrowths of the freshwater sponge Corvomeyenia carolinensis Harrison were examined using histological and histochemical techniques, supplemented by phase contrast observations of cellular behavior. The tissue and cellular components of the spongillid outgrowth region were defined in terms of function and morphogenic state. Archeocytes differ considderably, in both histochemical and morphological characteristics, from other cell types of the adult sponge, being histochemically similar to stem cells reported from a variety of developmental series. Archeocytes exhibit cytological characteristics of unspecialized cells capable of high levels of synthetic activity while other cell types of C. carolinensis, for the most part, can be characterized as fully differentiated cells displaying more restricted synthetic capabilities but often accumulating neutral mucoproteins. The presence of aggregates of amebocytes, not identifiable as archeocytes and possibly engaged in gemmule formation, is discussed in terms of current concepts of gemmulation and cellular developmental capabilities in sponges.

Notes: The long antennal flagellum of Neoconocephalus ensiger is covered with many sharp-tipped hairs that appear to be non-innervated; thick-walled chemoreceptors, that may also have a tactile function; thin-walled chemoreceptors of several kinds and coeloconic chemoreceptors. All of the chemoreceptors are innervated by small groups of neurons. The first flagellar subsegment is unusual in that it bears a small protuberance on its latero-ventral surface. This marks the site of the attachment, internally, of a scoloparium containing about eleven scolopales in which the dendrites of some 23 sensory neurons terminate. The most distal subsegment lacks the scoloparium reported earlier for the grasshopper. No conspicuous difference between the antennae of males and of females was found.

Notes: The antennae of two species of thrips, Bagnalliella yuccae (Hinds) and Frankliniella tritici (Fitch), have been examined with the light and electron microscopes. The antennal flagellum of both species is provided with tactile hairs, thick-walled chemoreceptors and thin-walled chemoreceptors. In addition, B. yuccae, but not F. tritici, has a single coeloconic chemoreceptor on the dorsal surface of the pedicel. Observations were made on the fluids in the lumen of the antennae of E. yuccae in the living insect. The movement of the fluids probably has an important physiological significance.

Notes: The cervicothoracic muscles of nymphal and adult Euborellia annulipes (Lucas) are described, the former for the first time. In the adult, eight new muscles are identified, while the nymphs possess a further seven muscles that disappear at maturation or before. Otherwise the same muscles occur in nymphs as in adults, though some nymphal muscles are less clearly separated from one another than their adult homologs. The attachment sites of certain muscles show a number of slight differences between nymphs and adults. The work emphasizes the necessity of taking the immature musculature into account in assessing the muscular pattern represented in an insect order.

Notes: Some of the cytological characteristics of the hemocytes of the scorpion, Palamnaeus swammerdami, were studied. The morphological features of the arachnid hemocytes were observed to be similar to those of hemocytes of insects, millipedes and isopods. Jones' system of hemocyte classification was extended to the arachnid hemocytes. The six classes of hemocytes indentified in the scorpion correspond to prohemocytes, plasmatocytes, granular hemoocytes, cystocytes, spherule cells and adipohemocytes of insects. A cell type comparable to oenocytoids of insects and crustaceans is absent. The prohemocytes can be subdivided into four categories that probably represent the precursors of the major cell types. The cytological characteristics of the major cell classes and the occurrence of the miniatures of some of these major cell types support the concept of Jones (62) that these cell types might have different cell lineages and might not be capable of transforming into one another. Some of the prohemocytes, plasmatocytes and granular hemocytes were amoeboid. The nature of the granules and the vacuoles of plasmatocytes and granular hemocytes were compared with the granules and vacuoles of corresponding hemocytes of other arthropods. Cystocytes did not bring about any visible coagulation similar to their counterparts in millipedes and crustaceans. Plasmatocytes, granular hemocytes and spherule cells were observed to occur in conglomerates. The cell types noted in the present study were compared with the hemocytes of other arachnids.

Notes: The morphological features of the hemocytes of the crustacean Ligia exotica are similar to hemocytes of insects and millipedes. Jones system of hemocyte classification is extended to crustacean hemocytes. As in insects, seven classes of hemocytes, identified as prohemocytes, plasmatocytes, granular hemocytes, cystocytes, oenocytoids, spherule cells and adipohemocytes, occur. The prohemocytes can be subdivided into five categories that probably represent the precursor of major cell types. The structural and chemical features of other major cell classes are distinct and support the concept of Jones ('62) that these types might have different lineages and might not be capable of transforming into one another. Some of the prohemocytes, plasmatocytes and granular hemocytes are amoeboid. Cystocytes do not bring about any visible plasma coagulation similar to their counterpart in millipedes. Oneocytoids and adipohemocytes are rare. Plasmatocytes, cystocytes and oenocytoids occur in conglomerates, the significance of which is discussed. The cell types are compared with those of the hemocytes of other crustaceans. It is suggested that the nomenclature based on morphological characters is more suited for crustacean hemocytes than a nomenclature based on behavioural and physiological characters.

Notes: Well preserved transitional cells were found between differentiated cells and horny cells of the frog epidermis, thus facilitating the study of the sequential events involved in horny cell formation. Autolysosomes appear to play an important role in the formation of horny cells. These structures preferentially digest those cytoplasmic components which are not necessary constituents of the terminal horny cell. The release of the contents of the small mucous granules into the intercellular spaces is one of the initial events in horny cell formation. Filaments and large mucous granules seem to be resistant to the lytic digestion and contribute to the bulk of the horny cell. Loss of fluids through the plasma membrane and consolidation of the remaining constituents, results in a flattened horny cell. The appearance of a thickened membrane around the horny cell signifies the completion of the transformation process.

Notes: The three dorsal ocelli of worker honeybees have been studied by light and electron microscopy. Each ocellus has a single flattened spheroidal lens and about 800 elongated retinular cells. Retinular cells are paired and form a two-part plate-like rhabdom between their distal processes. Each rhabdomere comprises parallel microvilli projecting laterally from the apposed retinular cells. Primary receptor cell axons synapse within the ocellus with ocellar nerve fibers of two different calibers. Each ocellus has eight thick fibers ca 10 m̈m in diameter and several thinner ones less than 3 m̈m in diameter. Fine structural evidence suggests that retinular axons end presynaptically on both types of ocellar nerve fibers. Since all retinular cells apparently synapse repeatedly with the thick fibers this involves a convergence of about 100:1. Thick fibers always terminate postsynaptically within the ocellus while thin fibers terminate presynaptically on other thin fibers, thick fibers or retinular axons. Structural evidence for synaptic polarization indicates that retinular cells and thick fibers are afferent, thin fibers efferent. Thus complex processing of the ocellar visual input can occur before the secondary neurons of the three ocelli converge to form the single short ocellar nerve which runs to the posterior forebrain.

Notes: The functional morphology of the forelimbs of the following African Viverridae was studied, Atilax paludinosus, Bdeogale crassicauda, Civettictis civetta, Genetta genetta, G. tigrina, Helogale parvula, Herpestes ichneumon, H. sanguineus, Ichneumia albicauda, Mungos mungo, Nandinia binotata. Their locomotory behaviour has been previously studied and described and is related to morphological differences. The osteology of all the species and the myology of three species is described. The species have been assigned to primary locomotor categories on the basis of their locomotion. These are 1, climbing, arboreal walking; 2, arboreal and terrestrial walking and jumping; 3, general terrestrial walking and scrambling; and 4, trotting. In the climbing arboreal walking category the most distinctive morphological adaptations are powerful flexors and extensors as well as a flexible plantigrade manus with retractile claws. In the arboreal and terrestrial walking category the shoulder, elbow and carpal joints are flexible and the manus has retractile claws, though the flexor and extensor musculature is insufficiently developed for controlled climbing. The trotting category is characterised by a high humero-radial index and a rigid antibrachium. The foot is digitigrade with the claws short and stout. Species in the general walking and scrambling category show many differences in the morphology of their feet, even though the proximal parts of the forelimb appear similar. Due to the restricted nature of the adaptations, these species have been assigned to secondary locomotor categories. Morphological characters typical of the locomotor categories are summarized in the discussion.

Notes: One of the main reasons for immobilizing an enzyme is to enable its reuse, or continuous use, in a reactor. Consequently immobilized enzyme stability is an important factor in enzyme reactor design. The performance of the reactor will decrease if during operation the support material disintegrates into smaller particles that pass out of the reactor system.When β-galactosidase is immobilized by covalent attachment to AE-cellulose, the smaller particles have a higher activity. After subjection of the immobilized enxyme to a shear stress the average particle size decreases and the total enzymic activity increases. A loss of small particles from the reactor, although constituting a small weight percent loss of support, will result in a disproportionately large loss in activity. The relevance of these observations to reactor performance is discussed.

Notes: A tubular membrane reactor offers many advantages over a solid wall reactor to carry out an enzyme catalyzed reaction. With proper membrane selectivity, the product, may be separated from the reacting stream and the enzyme recycled for continuous reuse. In most cases, enzyme reuse contributes to the economic feasibility of a continuous enzyme catalyzed process. Furthermore, the efficiency and performance of a membrane reactor is greater than that of a solid wall reactor.Continuous hydrolysis of starch by the enzyme β-amylase, carried out in a commercially available tubular membrane unit, is studied at different starch and enzyme concentrations for a given system pressure and inlet flow rate. Results show that the performance of the membrane reactor is in all cases greater than that of the solid wall reactor. A steady state in performance of permeation rate is, however, not reached by the membrane reactor, which shows a continuous decline within the periods examined in this study. This decline is caused in part by the aging of the starch solution, but mostly by the formation of a concentrated, or gel, layer at the membrane surface. This appears to be the main limiting factor for this process since the decline in reaction and permeation rate results in a severe decrease in the amount of maltose in the permeate.

Notes: The disintegration of baker's yeast (Saccharomyces cerevisiae) by a high pressure homogenizer, to a pressure of 25,000 psi. (172.37 MNm-2) is described, together with details of the methods of measurement used to obtain information on the valve movement and pressure transients. The theory of the mechanism of cell disintegration is discussed.

Notes: The mixing of the anaerobic digester contents significantly influences the efficiency of this operation; in particular, hydraulic dead zones are extremely detrimental to the reaction kinetics involved in anaerobic digestion. An analysis of the relative importance of thermal fluid movement in the digester to those caused by fluid inflow and outflow is presented. As an example, these principles are applied to a digester at the South Bend Wastewater Treatment Plant. Experimental measurements, which have general applicability for the measurement of digester mixing volume, confirm the theoretical conjectures. Various types of optimizations can be attempted on this mixing operation. One such optimization applied to gas lift mixers, as employed in the South Bend Treatment Plant, is illustrated.

Notes: Activated sludge is a widely used aerobic biological waste-water treatment process. A rational approach to least cost design of an integrated system is described which includes the following processes: activated sludge reactor, final settling tanks, gravity thickening, and aerobic sludge digestion. Both capital and operation and maintenance costs are considered. Biological reactor design is based on microbial kinetic concepts and continuous culture of microorganisms theory. Biological solids retention time (θc) is utilized as the primary independent design variable to which system performance is related, e.g., effluent quality, ammonia oxidation, and excess sludge production. Liquid-biomass separation is based on the batch flux technique, a rational approach to design of gravity separators (final settling tanks). Trade-offs among reactor volume, clarifier size, recycle pumping capacity, thickener capacity, digester volume, air requirements, and sludge production are discussed. The optimum design is taken as the combination of these parameters within the acceptable design domain, determined by effluent quality criteria, that results in minimum cost. While the method described is general, design of a given treatment system depends on availability, from lab or pilot studies, of system specific numerical values for biological growth coefficients and biomass setting characteristics. A design example illustrates the approach.

Notes: It has been demonstrated that ozone dosages of 0.06 lb and 0.78 lb per pound of lignin can reduce the optical density (in the visible range) of the latter by 82% and 92%, respectively. The reduction in color is accompanied by a shift in the molecular weight distribution of lignin from a broad peak of between 20,000 and 70,000 to lower molecular weight, species including substantial amounts possessing a molecular weight of 1000 or less. The cost of decolorizing a typical kraft paper bleach effluent is estimated to be under 50¢/1000 gal which compares favorably with competitive decolorizing processes. Lignin ozonation results in the production of a series of decolorized products which can serve as the sole source of carbon for a variety of microorganisms. Feasibility studies indicated that at least 40% of the ozonated material can be transferred into microbial biomass (protein) as well as other products of commercial interest such as fumaric acid and penicillin.

Notes: The consideration of fermentation principles in the treatment of industrial and sanitary waste waters leads to substantial process improvements. In particular, the rate of reaction can be improved by several fold by establishing the proper environmental conditions for microbial growth in a fermentation system. Recent work on the concentric cylinder air lift has shown it to be an economical fermentor with many advantages over conventional fermentors. An attempt to improve the economical performance of this system led to the development of the thin channel rectangular air lift fermentor. This was based upon a theoretical analysis of performance parameters. The analysis indicates decreased bubble coalescence, increased bubble entrainment, decreased power costs, increased mass transfer coefficients, and decrease capital costs. Experience using a prototype thin channel rectangular air lift system with sanitary and industrial waste-water treatment systems has demonstrated exceptionally high rates of BOD removal at low operating and capital costs.

Notes: A tubular loop batch fermentor has been designed and constructed, and was found to behave in a similar manner to a conventional stirred tank reactor. It appeared that foaming could be greatly reduced as no air ever encountered the impeller. The fluid mechanics of pipe flow are considerably simpler than tank flow patterns. On this basis a design procedure for a large scale tubular fermentor was outlined, which had considerable advantages over the more complex scale-up problems of a tank fermentor.

Notes: System response data for step changes in input tracer concentration have been obtained for two different impeller agitated continuous flow mixing systems containing aqueous polysaccharide solutions. The vessel volumes were 1.6 and 10.9 liters. Polysaccharide concentration, dilution rate, and impeller speed were varied according to a plan devised using dimensional analysis and assuming that bulk motion is the predominant mass transport mechanism in the system. The data show that this is not true and that serious errors may occur if scale-up calculations are based on assuming that bulk motion predominates. Under the operating conditions used, perfect mixing was not observed.

Notes: A column reactor with an annular cross section was formed by rolling up DEAE cellulose paper and a screening spacer. Glucoamylase was attached by ion adsorption. For the spacer used, pressure drop was very low, suggesting that this form may be useful with feed streams that are not completely particle-free. Tests of this reactor at the high substrate concentrations characteristic of commercial reactors showed very little diffusional resistance, exhibiting zero-order behavior over most of the concentration range. At low concentrations, the reactor had an apparent “half-order” behavior caused by diffusional limitation in the paper. In this range, flow rate influenced the reaction rate, showing that mass transfer in the main stream also is a contributing factor in this range. Because of the high concentrations and the low Michaelis constant (0.0011 M) the reactor does not show first-order behavior, even at very high conversions. The design of a plant-scale reactor was formulated from these data. The increase in the quantity of enzyme necessary to compensate for the effects of diffusion was only a few percent.Two reactors were formed with sheets nonporous to the enzyme, binding the enzyme with cyanogen bromide after forming the reactor. The amount of enzyme bound was about one monolayer, and there appeared to be no diffusional limitations, even at low substrate concentrations.

Notes: A high molecular weight intracellular enzyme of Bacillus brevis ATCC 9999 is released when the organism is disrupted by sonication of homogenization. However, both processes also degrade the enzyme. Assays for protein release and specific enzymatic activity of the released protein indicate that both release and degradation can be represented by first-order kinetic models. Utilization of the difference between the kinetics of release and degradation allows optimization in the recovery of this enzyme for both the sonication and homogenization processes.

Notes: Several microorganisms having higher L-histidine ammonia-lyase activity were immobilized into polyacrylamide gel lattice. The yield of enzyme activity by immobilization was highest in Achromobacter liquidum IAM 1667. As A. liquidum has urocanase activity, the cells were heat-treated at 70°C for 30 min to inactivate the urocanase.Enzymatic properties of the immobilized A. liquidum cells were investigated and compared with those of the intact cells. No difference was observed between the pH activity curve and optimal temperature for the intact and immobilized cells. The permeability of substrate or product through the cell wall was increased by immobilization of the cells.When an aqueous solution of 0.25M L-histidine (pH 9.0) containing 1mM Mg2+ was passed through a column packed with the immobilized A. liquidum cells at a flow rate of SV = 0.06 at 37°C, L-histidine was completely converted to urocanic acid. The L-histidine ammonia-lyase activity of the immobilized cell column was stable over 40 days at 37°C. From the effluent of the immobilized cell column, Urocanic acid was easily obtained in a good yield.

Notes: An apparatus is described which is designed for preparative freeze concentration experiments by the technique of normal freezing. It has a capacity of approximately 22 liters distributed over twelve vessels. The influence of various geometrical and chemical parameters such as stirring speed, crystallization rate, and sample composition on the normal freezing of protein solutions are discussed. For dilute protein solutions (〈0.1%) the concentration factor generally was 8- to 10-fold with recoveries of 90-100 percent. With higher protein concentrations and at ionic strengths higher than approximately 0.05, the recovery was decreased. No loss of activity was detected when concentrating enzyme solutions.

Notes: β-Amylase (EC 3.2.1.2), obtained from barley, was chemically attached to a crosslinked copolymer of acrylamide-acrylic acid using a water-soluble carbodiimide. The derivative showed 23% β-amylase activity in relation to that of free enzyme with a coupling yield of 40% based on the amount of added β-amylase. In order to find optimal coupling conditions, the effect of pH and different carbodiimide concentrations was investigated. The enzymic activity associated with different β-amylase concentrations was further outlined. A slightly increased operational stability for the enzyme upon immobilization was observed. Markedly improved operational stability has been obtained by coupling in the presence of reduced glutathione of bovine serum albumin.

Notes: Glucose oxidase (EC 1.1.3.4, from Aspergillus niger) has been entrapped in a crosslinked 2-hydroxycthyl methaerylate gel containing 20% poly(vinyl pyrrolidone). The kinetic behavior and thermal stability of the entrapped enzyme were found to closely approximate that of the free enzyme. The entrapped glucose oxidase shows a broadened pH profile which is attributed to a buffering effect of the gel. Stability of gel entrapped glucose oxidase is extremely good at room temperature, suggesting a variety ofanalytical and control uses for this system.

Notes: The effects of wall growth are described for a mixed methane-utilizing bacterial population growing in both batch and continuous culture. These effects are similar to those predicted previously by a theoretical analysis (Topiwala and Hamer, 1971).

Notes: A new and simple method for immobilization of enzymes by the aerobic radio-polymerization of acrylamide was developed. Irradiation treatment of acrylamide in the frozen state produces a spongy immobilized enzyme membrane without the addition of carriers. Aerobic polymerization yields of acrylamide in the frozen state were increased by the addition of starch and also by lyophilization.Glucose oxidase (activity recovery was 12.3-33.7%), invertase (69.2%), D-amono acid oxidase (25.0-70.5%), aminoacylase (39.2-43.7%), mold α-amylase (18.0%), malt β-amylase (4.1%), glucoamylase (6.5%), alkaline protease (5.3%), and neutral protease (10.5%) were immobilized by this method. Invertase entrapped by this method had a wider optium pH range and was active at higher temperatures.

Notes: Spirulina maxima, a high protein alga, was grown in effluents from the London municipal waste treatment plant. Optimum growth conditions were developed, the composition of algae and the removal of nitrogen and phosphorus in effluents were studied. The advantages of this process in tertiary waste-water treatment and the quality of the single cell protein were investigated.

Notes: Studies were carried out on the production of Sindbis, influenza and vesicular stomatitis viruses in suspensions of chicken embryo and rat embryo cells. The yield of Sindbis virus in chicken embryo cell suspensions was independent of the multiplicity of infection over the range 0.0001 to 0.01 although reduction in multiplicity caused a delay in virus production. With influenza virus the use of higher multiplicities gave increased virus yields possibly due to the very slow production at low multiplicities. In both monolayer and suspension cultures of chicken embryo cells addition of serum or use of media richer than minimum essential medium (Eagle) had little effect on Sindbis virus production, but if the glucose were omitted the virus yield was markedly reduced. In cell suspensions, a marked reduction in virus yield occurred if infection were delayed more than 24 hr after cell preparation whereas in monolayers the delay of infection allowed cell propagation and hence a higher yield of virus. It was also shown that vesicular stomatitis virus can be produced in chicken embryo cell suspensions, and that rat embryo primary cell suspensions can be used to prepare both Sindbis and vesicular stomatitis viruses. Sindbis virus obtained from chicken embryo cell suspensions was purified by polyethylene glycol precipitation and sucrose density gradient centrifugation and shown to contain only those proteins previously identified as viral, without any contamination from chicken cell proteins. The relative ease and economics of virus production by cell suspension and monolayer methods is compared.

Notes: Numerical solutions to the equations describing simultaneous mass transfer and enzymic reaction within porous spherical particles have been used to examine the effect of enzyme content and other parameters on the kinetic behavior of immobilized enzymes. These solutions have also been compared with experimental data for enzymes immobilized to DEAE-cellulose particles. The influence of particle size and enzyme content on catalyst design is discussed.

Notes: Degradation of UNA by endogenous RNase in cell suspensions of Saccharomyces cerevisiae was found to be achieved by mechanical disintegration followed by incubation in the presence of NaCl. The incubation parameters pH, temperature, time, and concentration of NaCl were investigated. Protein concentrates with a low content of RNA were obtained by precipitation of the incubated suspensions and separation of the degradation products.On a pilot plant scale the incubation was performed at 50°C and pH 5.6 in the presence of 3% NaCl for 20 min. Kilogram quantities of protein concentrates containing 1.4% RNA and 8.2% nitrogen were obtained. The RNA reduction and the nitrogen yield was 85 and 60%, respectively. The yield of amino acids was about 75%. The process described can probably be applied for large-scale production.

Notes: When the effect of catabolite repression is eliminated Saccharomyces cerevisiae prefers an aerobic metabolism. The potential for completely aerobic catabolism exists even in circumstances where its action is limited by the oxygen available. When the oxygen absorption in the medium is adequate, yeast uses a solely oxidative metabolism for energy-yielding reactions. The changes observed in the activity of malate dehydrogenase can be described as a function of two isoenzymes, both of which are affected by oxygen; the isoenzyme participating in the glyoxylate cycle shows variations in activity similar to that observed in isocitrate lyase. NAD-linked glutamate dehydrogenase activity roughly follows that of malate dehydrogenase and isocitrate lyase; in cultivations with the same growth rate the NADP-linked dehydrogenase is insensitive to the oxygen level. The cytochromes aa3, b, and c have a clear maximum at low oxygen tension, the most sensitive being cytochrome aa3. The imbalance between cytochrome c:oxygen oxidoreductase activity and the amount of cytochrome aa3, and the correlation observed between respiration rate and the activities of cytochrome c oxidase and NADH2:cytochroine c oxidoreductase are discussed. Methods used for estimation of cytochromes are compared.

Notes: An electronic particle counter has been used to estimate the disintegration by freeze-pressing baker's yeast. A counter threshold level which just yielded the maximum count for intact cells was selected. The conductivity of the suspending medium was chosen such that maximum counts were obtained. Under these conditions, the electronic counts agreed well with the visual counts. At a certain threshold level the maximum count was obtained at a lower resistivity (higher conductivity) in the suspending solution with the freeze-pressed suspension than with untreated cells, indicating that damage to the permeability barrier may occur without disruntion of the cell envelope.Fresh baker's yeast cells do not behave as nonconducting particles. This has to be taken into account when volume determinations with electronic particle counters are performed.

Notes: The soluble components in disintegrated cells of Saccharomyces cereivisiae have been characterized by means of extraction, centrifugation, dialysis, and gel filtration. The influence of alkali and heat treatment on the protein and RNA in the soluble fraction from disintegrated yeast cells and on functional properties of protein concentrates have been studied.After water extraction and centrifugation at 100000 g 42% of the nitrogen containing components of the disintegrated cells were recovered in the supernatant. By extraction at pH 11.5 an additional 31% of the nitrogen was solubilized. Half of the water-soluble nitrogen-containing components has a molecular weight lower than 5000. In the water- and alkali-soluble fractions about 80% of each amino acid was recoveredThe water-soluble protein was separated into 3 fractions by gel filtration on Sephadex G 200. The major portion of the protein had a molecular weight about 100,000. The amount of protein in this fraction was decreased after treatment at increasing pH and temperature. No degradation of protein to low molecular peptides occurred. The amount of RNA in the soluble fraction was only slightly influenced by alkali treatment and by heat treatment at pH 7.5 in the presence of 5% NaCl. RNA was not degraded to low molecular components of the treatments.The solubility of protein concentrates decreased after treatment at alkaline pH and after heat precipitation.

Notes: A simple system for the control of oxygen transfer in laboratory fermentation units has been devised. This system employs pure oxygen to elevate the driving force for oxygen transfer. In addition, the aeration is controlled in a closed loop manner in order to minimize total exit flow of the gas from the fermentor. Fermentation study using Bacillus stearothermophilus grown on n-dodecane at 55°C illustrates excellent carbon recovery as well as the ease of controlling oxygen transfer.

Notes: β-Galactosidase has been purified from an ammonium sulfate precipitate of E. coli strain ML308 by biospecific adsorption on a column of agarose gel substituted with p-aminophenyl-β-D-thiogalactopyranoside. The system described using a 1.8 liter column has a useful processing capacity of 3.8 × 106 units of β-galactosidase per 2 hr cycle. This corresponds to about 5 g of pure enzyme. An electromechanical timing device operates a set of six solenoid valves and carries out a preset program consisting of sample application, washing, and elation operations.

Notes: Porous hollow cellulose fibers have been used to separate a nonflowing enzyme solution of alkaline phosphatase from a continuous flow of substrate. The porosity of the hollow fiber membrane allows the substrate and product to diffuse freely through the membrane while restricting the permeation of the enzyme. The resulting “immobilized” enzyme system has been shown to behave as a continuous reactor - converting p-nitrophenylphosphate to p-nitrophenol. By varying the concentrations, flow rate, etc., either diffusion or enzyme kinetics can be studied. The continual influx of product and removal of substrate at steady state allows the study of kinetics of relatively short half-life enzymes and unstable systems.

Notes: The growth of a mixed methane-utilizing culture in a continuous flow fermenter has been studied under both methane and oxygen limitation. Small additions of methanol have been shown to inhibit the methane-utilizing moiety in the culture and it has been shown that the Hyphomicrobium sp. in the mixed culture removes any inhibitory methanol. The interaction between the methane-utilizing Pseudomonas sp., and the Hyphomicrobium sp. has been explained and a model of the continuous mixed culture under oxygen limitation has been formulated. Qualitative predictions of transient phenomena by the model have been verified experimentally.

Notes: L-Asparaginase has been attached by chemical means to the inner surface of nylon tubing. An experimental study has been carried out of the flow kinetics for such a system, asparagine solutions at various concentrations being passed through two lengths of tubing at various flow rates. Measurements were made of the concentration of the product ammonia at the tube exit, and of the rate of formation of ammonia, under the various conditions. Apparent Michaelis constants, Km(app), were some three orders of magnitude higher than the Km for the enzyme in free solution (∼13 × 10-6JM). The results were analyzed with respect to the theoretical treatment described in the preceding paper (Kobayashi and Laidler), three different methods being employed. It is concluded that at lower substrate concentrations and flow rates the reactions are largely diffusion-controlled, the enhanced Km(app) values being largely if not entirely due to the diffusion control; ionic strength studies showed electrostatic repulsion effects to be unimportant. At high concentrations and high flow rates (when the diffusion layer is of negligible thickness) the diffusional effects are minimized, and Km(app) approaches the true Km value for the immobilized enzyme.

Notes: An α-galactosidase which will hydrolyze the oligosaccharides melibiose, raffinose, and stachyose has been immobilized on nylon microfibrils suitable for use in large flow-through reactors. This catalyst system is stable for many months, both under use and storage conditions. The immobilized enzyme behaves similarly to the enzyme in solution, characteristically exhibiting both product and substrate inhibition. The catalyst is prepared in situ and a large, 8-liter reactor has been made. The catalyst has been used to reduce the raffinose concentration in beet sugar molasses.

Notes: Evolutionary operation (EVOP) was used to experimentally investigate the optimum steady state operating conditions for a step aeration activated sludge waste treatment process. A laboratory scale two tank step aeration activated sludge unit with fixed total volume, total influent flow rate, recycle flow rate, and sludge wasting rate was employed. The volume ratio and flow rate ratio which minimized effluent chemical oxygen demand were determined. The results indicate that EVOP is a useful technique for improving the performance of biological processes.

Notes: Trypsin (EC 3.4.4.4) was immobilized in low yield on aminoalkylsilylated magnetite (Fe3O4). Better results were obtained when trypsin was immobilized by crosslinking with glutaraldehyde on magnetite. The preparation contained 36 mg protein/g magnetite and the enzyme retained 46% and 11% of esterase and proteolytic activity. Immobilized trypsin was more heat stable than trypsin. Invertase (β-D-fructofuranoside fructohydrolase, EC 3.2.1.26) was cross-linked on magnetite with glutaraldehyde in low yield due to the inactivation of the enzyme. However in the presence of 1% sucrose, the total activity recovered was 79% of the initial activity and the preparation contained 4.4 mg/g of active invertase. Immobilized invertase was less active than invertase when acting on oligosaccharides of the raffinose family. The immobilized enzymes could be easily recovered, from solutions or suspensions, magnetically.

Notes: A general model of the kinetics of microbial growth has been developed involving the kinetics of incorporation of substrate into biomass and the maintenance energy requirements.Results obtained from batch cultures of the yeast Saccharomyces cerevisiae growing in synthetic media at pH 5.1 and 30°C permitted all biological parameters in the model to be calculated.Values obtained for these parameters were: maximum specific glucose uptake rate (μSm), 2.08 g/g biomass/hr; apparent Michaelis constant for glucose (KS), 0.1 g/liter (5.5 × 10-4M) apparent Michaelis constant for oxygen (KL), 1.4% O2 (3.2 × 10-6 M) quantitative index of the Pasteur effect (b), 4.9 × 10-4%-1 O2 (207 M -1). Under conditions of strongly substrate-repressed respiration the values obtained for YATP and P/O were constant over the course of the exponential phase of growth (YATP = 10.4 g biomass/mole ATP; P/O = 3 moles ATP/atom 0). Mass balances for aerobic and anaerobic cultures confirmed the results obtained form the generalized model. Results presented suggested the operation of a mechanism for regulating energy-yielding metabolism which involved an equilibrium between the systems of oxidative phosphorylation and dephosphorylation and was dependent upon the level of catbolite repression.

Notes: Invertase was entrapped in cellulose triacetate fibers and the properties of the insoluble derivative were studied. Fiber-entrapped invertase was found very stable under operating conditions. For some insoluble preparations a half-life value of 5,300 days was calculated; a sample of invertase fibers, continuously hydrolyzing sucrose, maintained unchanged its activity for five years. The activity displayed by invertase fibers was 15-65% of that of the free enzyme, depending on the amount of entrapped enzyme and on the porosity of the fibers. At very high substrate concentrations the activity of the entrapped invertase approximated to that of the free enzyme. The pH optimum for activity was around 4.5 for the free and entrapped invertase. The native and entrapped enzyme was unstable at temperatures higher than 35°C.The continuous hydrolysis of sucrose using invertase fiber was studied and the potential industrial application of entrapped enzyme is discussed.

Notes: An immunoadsorbent process has been devised for removing serum hepatitis antigen (HAA), from blood, blood plasma, and plasma products.The immunoadsorbent complexes specifically with HAA and the complex is removed from the plasma by filtration. The complex is dissociated with 0.23M NH4OH. The immunoadsorbant is regenerated for further processing, while the purified HAA by-product my be used to produce more antibody in animals.The rate of complexing is such that HAA is reduced one log cycle each 2hr. Since available tests can only detect HAA to about 109 particles per ml, it is proposed that HAA negative plasma and plasma products can be processed to reduce HAA to a probability of less than one HAA particle per plasma pool.A gibbon injected with infectious commercial Factor IX that was subjected to the immunoadsorbent process showed no sign of infection after 8 months. However 14 weeks after injection with unprocessed Factor IX, the gibbon showed signs of infection.

Notes: The time course of release of intact labile material from cells by cavitating ultrasound is examined. Curves are presented which show the yield of intact subcellular components obtainable from cells as a function of sonication time for various values of the ratio of inactivation rate to release rate. The general cases where inactivation of released product is concentration dependent (chemical) and concentration independent (mechanical) are considered.For a flow system the time of attainment of equilibrium concentration of active product is analyzed as a function of flow rate, release rate, inactivation rate, and volume of chamber. Curves of optimal yields to be expected for batch and flow systems are presented. It is shown that sonochemical inactivation can be made negligible by sonication of high cell concentration suspensions.

Notes: Among the methods used for the rupture of cells by means of solid shear, the Hughes press is best known. It operates by forcing an aqueous suspension of cells previously frozen at -27°C under pressure through a narrow circumferential gap. The mode of action requires a knowledge of the isentropic compression of the solid phase and its possible involvement with a phase change from ice I to ice III. It is shown that the mechanical properties of the solid mixture are relevant. A model is proposed of the plastic flow for the geometry of the press and it is shown that cell breakage is proportional to the velocity of the plunger and inversely proportional to gap width.

Notes: The CO2 mass balance for a suspension of algae cultivated in the region of neutral pH of the suspension on an open cultivation surface with a flowing suspension was assessed. From this balance an analytical expression for the course of free CO2 dissolved in the suspension along the flow of the suspension is obtained. Relations for the rate of CO2 decrease in the suspension and the utilization of CO2 supplied to the cultivation surface are derived.

Notes: Films of highly polymerized collagen, prepared in industrial conditions, were chosen for surface covalent binding of enzymes because of their insolubility, mechanical resistance, proteic nature, hydrophilic properties and for their abundance in chemically activable —COOH. Untanned films, previously acid- methylated, were activated by acyl azide formation. After removal of reagents by repeated washing, the coupling of enzyme was performed by immersion of the activated film in the enzyme solutions (2 to 3 hr, 0°C). The procedure is particularly mild since the enzymes never come into contact with chemical reagents, and thus avoid all denaturing processes. All the enzymes tested were successfully bound: glutamate dehydrogenase, lactate dehydrogenase, malate dehydrogenase, aspartate aminotransferase, glutamate pyruvate transaminase, creatine kinase, hexokinase, trypsin, and urease. As tested with aspartate amino transferase, enzymatic activity remained constant for months (100% after 5 months) in spite of repeated use of the film at 30°C, washing and storage in buffer at 4°C between assays.

Notes: Highly charged water-soluble conjugates of amyloglucosidase with a copolymer of ethylene-maleic acid or styrene-maleic acid were prepared and adsorbed on DEAE-cellulose and other cationic resins. Negatively charged enzyme conjugates were also obtained by means of succinylation. The polyanionic conjugates of amyloglucosidase adsorbed on the cationic carriers show higher temperature stability than the native enzyme adsorbed on the same carriers under the same experimental conditions. The insoluble enzyme conjugate-carrier complexes could be reused and retained their full activity while working continuously for three weeks. The application of the insoluble enzyme preparations with the above characteristics which could possibly satisfy the requirements for industry are discussed.

Notes: Ribulose-1,5-diphosphate carboxylase from spinach has been bound to arylamine porous glass with a diazo linkage and to alklamine porous glass with glutaraldehyde. Stability at elevated temperatures and responses to changes of pH and ribulose-1,5-diphosphate, Mg2+, and dithiothreitol concentrations were not significantly different from the soluble enzyme, though stability at 4°C was somewhat improved.

Notes: Continuous fermentation of grape juice was studied in three basic kinds of nonmechanically stirred fermentors, classified according to the direction in which flow is applied against settling: upward-flow, slant-flow, and horizontal-flow.Fermentation kinetics were found to be zero-order, thus rate was directly proportional to cell density.Completeness of fermentation depends upon yeast characteristics (growth rate flocculation), fermentor geometry (settling depth, flow path length), and upon flow velocity.Fermentor analysis demonstrated that to achieve complete fermentation in the minimum juice holding time, efficient cell retention as well as rapid yeast growth and fermentation rates are required. Fermentor design and operation are discussed in the light of this analysis.

Notes: Automated sampling device was designed to permit determination of rates of biological transport of metabolites into cells. The substrate was automatically introduced into a stirred cell suspension at 37°C. The first sample was automatically taken after a mixing interval of 1 sec and nine subsequent samples were taken at programmable intervals (1 to 100 sec). The samples were forced by pressure differential (vacuum) through 0.4 μ pore size membranes and approximately 50 μl were collected in disposable cups. The duration of the sampling interval was controllable down to 0.1 sec. The samples preserved records of the substrate concentrations in solution at the time of filtration. With the use of suitable radioactive labeled isotopes, the changes in substrate concentrations may be conveniently measured by liquid scintillation spectrometry, but other analytical procedures of suitable sensitivity may be used. Initial and steady-state transport rates of succinate and glucose in Escherichia coli were obtained using the device.

Notes: A theoretical treatment has been worked out for the kinetics of solid-supported enzyme systems, with diffusive and electrostatic effects taken into account. A utilization factor, defined as the ratio of the actual reaction rate to the rate of substrate consumption in the outer solution, is defined, and equations to evaluate the utilization factor are given for five kinetic conditions: (a) Michaelis-Menten behavior, (b) substrate inhibition, (c) product inhibition (competitive), (d) product inhibition (noncompetitive), and (e) product inhibition (anticompetitive). When the solid-supported enzymes obey a Michaelis-Menten relationship, an equation for the apparent Michaelis constant is given and a criterion for insignificant diffusion effects is shown. A substrate-inhibited enzyme reaction may display multiple steady-state behavior, and a criterion for uniqueness is presented. In the case of product-inhibited enzyme reactions, the utilization factor is always less than that which corresponds to a Michaelis-Menten relationship. Equations to evaluate the apparent Michaelis and inhibition constants are given.

Notes: HeLa S3 cells in suspension culture are resynchronized in each successive generation by exposure to 0.25mM thymidine. The resynchronization approach appears to be superior to single- and double-block techniques because the period of effective inhibition of DNA synthesis is significantly shortened. Gram quantities of highly synchronous viable cells have been produced on a daily basis.

Notes: Formose syrup was studied as a carbon source for growth of a series of microorganisms obtained from various collections. Approximately 80 strains of bacteria, yeasts, and molds were inoculated into a medium containing formose syrup and mineral salts supplemented with small amounts of yeast extract and casein hydrolysate to supply accessory growth factors. Two preparations of formose syrup, produced by two different laboratories, were employed. Formose syrup I, characterized by a low sugar content, was poorly utilized; syrup II, containing a higher sugar concentration, was utilized to a greater extent. Two strains of Aerobacter acrogenes yielded 1.3 g dry cell mass from an initial charge of 10 g of formose II solids, whereas growth on 10 g of D-glucose amounted to 3.7 g.Klebsiella aerogenes MIT-B44, the best microbial strain isolated from soil by an enrichment technique, produced 1.3 g cells from 10 g fromose syrup II solids in supplemented medium; in direct comparisons, it produced 10-15% more cell 0.7-0.9 g cells per 10 g formose and grew with a doubling time of 55-70 min. Under such conditions, its macromolecular composition was 52% protein, 22% RNA, and 2% DNA. Although the apparent yield of cells from formose was only 8-11%, the actual yield based on formose utilized was 30%, the same as observed with glucose.A second strain was isolated from soil by enrichment with spent broth from K. aerogenes. This unidentified gram-negative, short rod-shaped bacterium grew in mixed culture with strain MIT-B44; in unsupplemented media they produced 1.55 g cells from 10 g formose II solids and 2.9 g cells from 10 g glucose.

Notes: Disintegration of microorganisms in a continuously working industrial homogenizer has been studied. The homogenizer consists of rotating discs in a cylinder filled with glass beads. Different parameters for disintegration of baker's yeast were investigated. The disintegration process is a first-order reaction and it is influenced by the flow rate of the suspension and by the agitator speed. At a flow rate of 200 liters/hr about 85% of the yeast cells can be disrupted in a single pass through the disintegrator. This type of disintegrator can be used for disruption of cells in order to produce single-cell protein, active enzymes and other valuable cell components.

Notes: The modeling of biological wastewater treatment processes has received much attention over the past ten years. Efforts are underway to develop a unified model for these processes which will greatly aid in the design and operation of wastewater treatment facilities. This paper presents a philosophical discussion of model building strategies augmented by a discussion of statistical problems associated with model parameter estimation and model discrimination. This discussion further illustrates with actual data, that goodness of fit is not a sufficient condition for model acceptance. Numerous rival models are examined to illustrate this point. In order to verify a model or to discriminate between rival models, they must be “Put in jeopardy.” If this attitude is not employed in model building efforts, important discrepancies in the proposed model may go undetected.