Cellulose digestion in termites and cockroaches: What role do symbionts play?

• 1.1. Termites and cockroaches are excellent models for studying the role of symbionts in cellulose digestion in insects: they eat cellulose in a variety of forms and may or may not have symbionts.
• 2.2. The wood-eating cockroach, Panesthia cribrata, can be maintained indefinitely, free of microorganisms, on a diet of crystalline cellulose. Under these conditions the RQ is 1, indicating that the cockroach is surviving on glucose produced by endogenous cellulase.
• 3.3. The in vitro rate at which glucose is produced from crystalline cellulose by gut extracts from P. cribrata and Nasutitermes walkeri is comparable to the in vivo production of CO₂ in these insects, clearly indicating that the rate of glucose production from crystalline cellulose is sufficient for their needs.
• 4.4. In all termites and cockroaches examined, cellulase activity was found in the salivary glands and predominantly in the foregut and midgut. These regions are the normal sites of secretion of digestive enzymes and are either devoid of microorganisms (salivary glands) or have very low numbers.
• 5.5. Endogeneous cellulases from termites and cockroaches consist of multiple endo-β-1,4-glucanase (EC 3.2.1.4) and β-1,4-glucosidase (EC 3.2.1.21) components. There is no evidence that an exo-β-1,4-glucanase (cellobiohydrolase) (EC 3.2.1.91) is involved in, or needed for, the production of glucose from crystalline cellulose in termites or cockroaches as the endo-β-1,4-glucanase components are active against both crystalline cellulose and carboxymethylcellulose.
• 6.6. There is no evidence that bacteria are involved in cellulose digestion in termites and cockroaches. The cellulase associated with the fungus garden of M. michaelseni is distinct from that in the midgut; there is little indication that the fungal enzymes are acquired or needed. Lower termites such as Coptotermes lacteus have Protozoa in their hindgut which produce a cellulase(s) quite distinct from that in the foregut and midgut.

     

Gas tensions within the egg of the quail (Coturnix c. Japonica) during the closing stages of embryonic development

• 1.1. The respiratory status of the embryonic quail during the two days prior to hatching was assessed by measuring the gas tensions within the air space of the egg and of blood collected from the chorioallantois.
• 2.2. When the lungs became inflated there was a significant decrease in the pO₂ of the gas in the air space.
• 3.3. After pipping, there was a rise in the pO₂ and fall in the pCO₂ within the air space, together with corresponding changes in the blood.
• 4.4. The outer shell membrane remained intact until the onset of hatching.
• 5.5. These results were compared with those obtained by other workers using the domestic fowl.

     

Thermal properties for digestive enzymes of a sub-antarctic beetle,hydromedion sparsutum (coleoptera,perimylopidae) compared to those in two thermophilic insects

• 1.1. Proteolytic, lipolytic, amylolytic and cellulolytic activities were studied in adults of the phytophagous beetle, Hydromedion sparsutum, indigenous to the sub-Antarctic island of South Georgia.
• 2.2. Gastric enzyme activities were measured at experimental temperatures of 5–40°C and results were compared with those obtained from two thermophilic insects, Gryllus bimaculatus and Tenebrio molitor.
• 3.3. Protease and lipase activities in Hydromedion were 10–15 times lower than in Gryllus and Tenebrio.
• 4.4. In the temperature range of 5–15°C, α-amylase activity from Hydromedion was only slightly lower than that from Gryllus.
• 5.5. Hydromedion gut homogenates exhibited a distinct cellulolytic activity, even at a low temperature of 5°C.
• 6.6. Cellulolytic activity in the digestive tract of Hydromedion was confirmed by the evolution of ¹⁴CO₂ after consumption of labelled cellulose.
• 7.7. The thermal properties of digestive enzymes agree well with the role of Hydromedion as primary decomposer in its ecosystem.

     

Respiration in the eastern water dragon,Physignathus lesueurii (agamidae)

• 1.1. Some aspects of the gas exchange system of a diving lizard, Physignathus lesuewii were studied.
• 2.2. Breathing patterns were analysed.
• 3.3. Breathing rate increases logarithmically with temperature and Q₁₀ = 1.8. LogBR = −0.237 + 0.0256 T.
• 4.4. Gas tensions in lung air and arterial and venous blood were measured. Arterial pH declines with increasing temperature.
• 5.5. Temperature has a marked effect on oxygen affinity of the blood (ΔH = −10.1 kcal mol⁻). A Bohr effect was also noted.
• 6.6. CO₂ equilibrium curves were drawn.
• 7.7. The results are considered with a view to anticipating the efficiency of the gas exchange system of this species under conditions of variable temperature and during diving.

     

The composition of cuticular hydrocarbons of the khapra beetles,trogoderma granarium

• 1.1. The cuticular and internal hydrocarbons of the khapra beetle Trogoderma granarium were studied by capillary column gas chromatography and mass spectrometry. n-Alkanes, 3-methyl-, 5-methyl-, 11-methyl-, 12-methyl-, 13-methyl-, 14-methyl- and 15-methylalkanes were found in the cuticular and internal lipids.
• 2.2. Some quantitative differences of the compositions were estimated for cuticular and internal hydrocarbons.
• 3.3. The n-alkanes in the samples are mostly odd chain lengths from 23 to 33 carbon atoms. In turn, the branched hydrocarbons consist of even carbon numbers ranging from C₂₆ to C₃₂, but the branching points are situated on the odd carbon numbers.
• 4.4. There are similarities in the n-alkanes patterns extracted from khapra beetle and wheat grains, the latter of which are the natural nutrition of this pest.

     

In vitro substrate utilization for lipid synthesis in liver explants from hyperthyroid chickens

• 1.1. Indian River male broiler chickens growing from 7 to 28 days of age were fed diets containing 12, 18, 24 and 30% protein + 0 or 1 mg triiodothyronine (T₃)/kg of diet to study energetic costs of lipogenesis and the use of various substrates for in vitro lipogenesis.
• 2.2. De novo lipid and CO₂ production were determined in the presence of [1-¹⁴C]pyruvate, [2-¹⁴q]pyruvate, [3-¹⁴C]pyruvate, [2-¹⁴C]acetate and [U-¹⁴C]alanine.
• 3.3. Oxygen consumption was determined in mitochondrial preparations to estimate the energetic costs in expiants synthesizing lipid.
• 4.4. Radiolabeled CO₂ derived from [1-¹⁴C]pyruvate was used as an estimate of coenzyme A availability in liver expiants. Lipids derived from [2-¹⁴C]pyruvate, [2-¹⁴C]acetate and [U-¹⁴C]alanine estimate relative substrate efficiency.
• 5.5. Labeled CO₂ production from [1-¹⁴C]pyruvate was greatest in that group fed a 12% protein diet and least in the group fed a 30% protein diet.
• 6.6. In addition, T₃ increased CO₂ production from [1-¹⁴C]pyruvate.
• 7.7. The production of ¹⁴CO₂ from the second carbon of pyruvate or acetate was increased by T₃.
• 8.8. The low-protein diet (12% protein) increased (P <0.05) lipogenesis.
• 9.9. Adding T₃ to the diets decreased carbon flux into lipid from all substrates, but increased CO₂ production from all substrates without changing stage 3 and 4 respiration rates in mitochondrial preparations.
• 10.10. These observations imply that coenzyme A availability may have regulated de novo lipogenesis in the present study.
• 11.11. It was also concluded that previously noted effects of T₃ on intermediary metabolism may involve metabolic pathways that do not involve changes in mitochondrial function.

     

Characterization of Colorado potato beetle lipophorin: A hydrocarbon-rich diacylgycerol-poor lipophorin

• 1.1. The lipid composition of lipophorin from the Colorado potato beetle, Leptinotarsa decemlineata Say, was analyzed.
• 2.2. This insect lipophorin contains 44% lipid and is characterized by large amounts of hydrocarbons and small amounts of diacylglycerol.
• 3.3. This is the first observation of a diacylglycerol-poor insect lipophorin in haemolymph.
• 4.4. Since the main energy source for flight in the Colorado potato beetle is proline, the low diacylglycerol content in lipophorin must be related to its peculiar flight metabolism.
• 5.5. This lipophorin, however, can still take up appreciable amounts of diacylglycerol from the locust fat body. Hydrocarbon uptake by this lipophorin was also demonstrated.
• 6.6. The main function of this lipophorin therefore seems to be transport of hydrocarbons from oenocytes to the cuticle.

     

The effect of temperature on the acid-base status of the blood of the hatching quail (Coturnix c. japonica)

• 1.1. The ambient temperature of embryos of pipped eggs was reduced from 38 to 28°C for a period of 45 min.
• 2.2. The blood P_CO2 was lower and the blood more alkaline at 28°C than at 38°C.
• 3.3. At 28°C plasma [HCO₃⁻] ] was lower than predicted from the blood buffer line determined in vitro.
• 4.4. The plasma concentrations of strong ions and lactate were the same at both temperatures.
• 5.5. After the ambient temperature had been returned to 38°C for a period of 45 min, blood pH was more acidic than before cooling, but there was no difference in blood P_CO2.
• 6.6. The plasma [HCO₃⁻] was the same as that at 28°C and plasma [K⁺] was higher than before cooling.
• 7.7. The results arc discussed in relation to the factors affecting blood pH in embryos at this stage of development.

     

A simplified method for assaying micromolar quantities of CO2

• 1.1. A rapid and inexpensive method is described for the determination of micromolar quantities of CO₂. which is well suited for infrequent analysis of 50 or fewer samples, and which lends itself for use as an undergraduate laboratory exercise.
• 2.2. The limits of resolution without elaborate precautions during sample preparation are 6 × 10⁻⁷ equiv, but simple modifications would yield a resolution of ca. 1 × 10⁻⁸ equiv.
• 3.3. The method involves placing a small fluid sample (10–100μ1) in the center well of a modified Conway type microdiffusion dish, driving off the CO₂ via acidification and trapping the released carbon dioxide in barium hydroxide located within the chambers middle well.
• 4.4. The quantity of CO₂ captured is determined by back titration, and bicarbonate levels extrapolated by application of the Henderson-Hasselbalch equation.

     

A comparative study on the biological properties of venoms from juvenile and adult common tiger snake (Notechis scutatus) venoms

• 1.1. The biological properties of venoms from juvenile and adult common tiger snakes (Notechis scutatus) were compared.
• 2.2. The lethality, procoagulant activity and enzymatic activities of the juvenile venom were not substantially different from those of the adult venom.
• 3.3. Electrophoretic studies, however, indicated some minor differences in the protein composition of the juvenile and adult venoms.

     

Drosophila mettleri and senita cactus alkaloids: Fitness measurements and their ecological significance

• 1.1. Drosophila mettleri have been found feeding but not breeding on decaying stems of senita cactus, the normal host for Drosophila pachea.
• 2.2. Alkaloids were extracted from senita stems and used in tests of egg-to-adult viability, developmental rate, and adult longevity.
• 3.3. The results show that developmental rate is not appreciably affected by senita alkaloids.
• 4.4. In general, D. mettleri was less affected by the alkaloids with respect to egg-to-adult viability and adult longevity than D. pachea at concentrations which are fatal to other desert Drosophila.
• 5.5. Tolerance to alkaloids gives D. mettleri an ecological advantage.

     

Metabolic rates of Calathus melanocephalus (L.) (coleoptera,carabidae) from alpine and lowland habitats (jeløy and finse,norway and drenthe,the netherlands)

• 1.1. Metabolic rates (ml O₂/mg/hr) of three geographically separated populations of the carabid beetle Calathus melanocephalus L. (Finse and Je 10y, Norway and Drenthe, The Netherlands) were measured and compared by ANCOVA.
• 2.2. No significant relationship (P > 0.05) between metabolic rates and body weight or sex of the animals were found.
• 3.3. Individuals mostly acclimated to low temperatures by increased metabolic rates and in the opposite direction to higher temperatures. Individuals collected in early summer also showed higher metabolic rates than those caught later in the autumn.
• 4.4. Contradicting the theory of metabolic cold adaptation, beetles from The Netherlands had the highest metabolic rates, beetles from Finse intermediate rates and beetles from Jeløy the lowest rates.
• 5.5. No significant relation were found between geographical origin of the beetles and their respective chill-coma temperature.

     

The effect of d2o on glycolysis by rat hepatocytes

• 1.1. The effect of incorporating D₂O into the incubation medium on glycolysis and gluconeogenesis by hepatocytes from fasted rats was examined.
• 2.2. The substitution by heavy water, D₂O, at concentrations from 10 to 40%, stimulated glucose uptake, lactate production and CO₂ yields from glucose. At 10 mM glucose, 40% D₂O doubled glucose uptake, increased CO₂ production by 40%, and increased lactate production by 350%.
• 3.3. The stimulation of lactate production decreased at higher glucose concentrations, but was still substantial even at 80 mM glucose.
• 4.4. There was no effect on CO₂ production above glucose concentrations of 30 mM.
• 5.5. Ten percent D₂O showed little inhibition of lactate uptake, its oxidation and gluconeogenesis. At 40% D₂O the inhibition ranged from 10 to 20%.
• 6.6. No effect of D₂O on the rate of glucokinase or glucose-6-phosphatase was observed.
• 7.7. The concentration of fructose, 2,6-P was not affected by D₂O

     

Respiratory properties of blood and hemoglobin solutions from the piranha

• 1. Respiratory properties of piranha blood are distinguished from those of other fish primarily by the high CO₂ buffering capacity (ΔHCO₃/⁻ΔpH= 19.6mmol/l for oxygenated blood and 39.1 mmol/l for deoxygenated blood).
• 2. The concentration of nucleoside triphosphates (NTP) and the half-saturation tension (P₅₀) of whole blood were found to be inversely related to body size.
• 3. The higherP₅₀ in smaller fish, analogous to values obtained in previous studies involving interspecies comparisons, could be adaptive to a higher weight-specific metabolic rate.
• 4. Both ATP and guanosine triphosphate (GTP) lowered the oxygen affinity of purified hemoglobin solutions, accounting for the size-dependent correlation ofP₅₀ and NTP concentration in whole blood.
• 5. While similar in concentration in red cells, GTP is more potent than ATP as an allosteric modifier of hemoglobin function.

     

Chemical control of breathing in the lizard,Varanus exanthematicus

• 1.1. In Varanus exanthematicus, alterations of inspired and end-tidal gas pressures resulted in rapid ventilatory responses. This, as well as other physiological characteristics, contributes to a homeostatic ability unusual for a reptile.
• 2.2. In contrast to the lizard Lacerta, the varanid showed a positive ventilatory response to hypoxia.
• 3.3. Whereas CO₂ breathing depressed breathing frequency in other lizards, both tidal volume and frequency increased in Varanus.
• 4.4. End-tidal P_co2 increased significantly after feeding, presumably compensating for an alkaline “tide” of digestion. The increased end-tidal P_co2 not due to a decrease in ventilation.

     

Functional studies on the single component hemoglobin from an Amazon knife fish,Sternopygus macrurus

• 1. The whole blood of the non-air-breathing gymnotid teleost,Sternopygus macrurus, is half-saturated with oxygen at 5.2 mm Hg (apparent value) at 30°C in the absence of CO₂. Addition of 5.6% CO₂ causes the apparentP₅₀ value to increase over 3-fold.
• 2. The oxygen affinity of the stripped single-component hemoglobin at 20°C increases about 20-fold between pH 5.8 and 8.6 in the absence of ATP. This difference increases to 100-fold in the presence of 1 mM ATP.
• 3. A substantial Root effect is present: the stripped hemoglobin is only 70% saturated with O₂ at pH less than 6 when equilibrated with air.
• 4. The value of the Hill coefficient,n, is maximal near pH 7.0–7.5, and approaches 1.0 at high pH. The value is about 1.5 at low pH in the absence of ATP and 1.0 in the presence of 1 mM ATP.
• 5. The O₂ dissociation kinetics are heterogeneous at all pH values but most heterogeneous at low pH. The rate increases substantially as the pH decreases.
• 6. The CO combination kinetics as measured by the stopped flow technique are largely homogeneous except at high pH, but the CO combination kinetics after flash photolysis are markedly heterogeneous.

     

Seasonal variations in the anaerobic metabolism of the mussel Mytilus edulis (L.)

• 1.1. The incorporation of atmospheric ¹⁴CO₂ into the ethanol soluble metabolites of the tissues of the mussel, Mytilus edulis, exposed to the atmosphere, was measured monthly between July 1976 and June 1977.
• 2.2. The total incorporation of ¹⁴CO₂ was found to be greater in the summer than in the winter and these seasonal effects were not temperature-dependent.
• 3.3. The incorporation of ¹⁴CO₂ into the individual metabolites is discussed in relation to possible seasonal variations in the operation of the anaerobic succinate pathway and seasonal variations of tissue glycogen concentrations.

     

End products of carbohydrate metabolism in Trichomonas vaginalis

• 1.1. The major excreted acidic end products of the anaerobic and aerobic carbohydrate metabolism of Trichomonas vaginalis (ATCC 30001) were acetate and lactate. Glycerol-1-phosphate, pyruvate, malate and ethanol were also detected in the incubation medium, but they represented less than 3% of the total carbon excreted. Succinate could not be found. Under anaerobic conditions H₂ and CO₂ were produced. Under aerobic conditions O₂ was consumed and CO₂ produced.
• 2.2. In the absence of exogeneous carbohydrate more acetate than lactate was produced. Glucose (50 mM) significantly increased acid production with lactate becoming the predominant product. Glucose also increased the anaerobic and aerobic gas exchange.
• 3.3. In the presence of 5% CO₂ there were no significant changes in carbohydrate utilization and acid production.
• 4.4. Aerobiosis was accompanied by increased carbohydrate utilization and end product formation. No Pasteur effect could be observed.

     

Bioelectrical potentials across the mantle epithelium of Achatina fulica

• 1.1. The shell side of the mantle of Achatina fulica is several millivolts positive to the blood side in vitro.
• 2.2. The electrical potential does not depend on Na⁺, Ca²⁺, Mg²⁺, K⁺ or HCO₃⁻ but requires the presence of chloride on the shell side.
• 3.3. The potential difference and short-circuit current ranged from 3.0 to 30.0 mV and 15.0 to 75 μA/cm² with averages at 10m V and 50 μA/cm² respectively.
• 4.4. The electrical gradient is reduced by 2,4-dinitrophenol, thiocyanate and furosemide but not by ouabain, CO₂ or acetozolamide.
• 5.5. It is suggested that the nature and mechanism of electrogenesis in Achatina parallels that of the Helix mantle.

     

Characteristics of Crassostrea virginica crystalline style chitin digestion

• 1.1. Fundamental chitin digestion characteristics of Crassostrea virginica crystalline style were investigated.
• 2.2. Optimum temperature and pH were 34°C and 4.8. respectively.
• 3.3. The colloidal regenerated chitin (0.56mol/0.5 ml: GlcNAc equivalents) was saturating under all enzyme levels encountered.
• 4.4. There was no evidence of end product inhibition, even after 100 hr incubation.
• 5.5. Calculated K_m for the chitinase complex was 1.19mM when determined using a 30 min assay, but was only 0.70 mM when determined using a 4.6 hr assay.
• 6.6. Both K_m values are lower than reported for similar assays in other molluscs and for most bacteria.
• 7.7. Effect of substrate preparation on the kinetics are discussed.
• 8.8. Eight peaks of chitinase activity were resolved by DEAE-Fractogel ion exchange chromatography.