Counter current immunoelectrophoresis, a new technique for the rapid serodiagnosis of porcine cysticercosis

Counter current immunoelectrophoresis was used to detect porcine cysticercosis and water soluble extracts of scolex and of scolex with cyst wall were used as antigens. Serum samples from 40 pigs infected with Cysticercus cellulosae, five infected with C. tenuicollis and five with hydatid cysts, and 15 healthy pigs were tested. A sharp and thick concave precipitin band was observed at the point of interaction of antigen and antibody within 90 min in 39 sera from infected pigs (97.5%). The precipitin reaction was better in barbitone buffer of pH 8.6 with the well distance at 6 mm. No false or cross reaction were found and the test was rapid and sensitive.     

On the interactions between pancreatic lipase and colipase and the substrate, and the importance of bile salts.

The interactions between pancreatic lipase and colipase and the substrate and the effect of bile salts on these interactions have been investigated by the use of kinetic experiments and studies on the semiquantitative phase distribution of lipase and colipase activities. The results suggest that lipase binds to hydrophobic interfaces with partial irreversible inactivation. Bile salts in the range of micellar concentrations and above a pH of about 6.5 displace lipase from this binding, resulting in a reversible in activation. At pH values below about 6.5, lipase binds strongly to the substrate even in the presence of bile salt, and a low activity peak is seen around pH 5.5. This is the result of the binding of lipase to the "supersubstrate" and the activity of the catalytic site. In the presence of bile salt, colipase promotes the binding of lipase to the "supersubstrate" but not to other hydrophobic interfaces, and catalytic activity is reestablished. Kinetic data indicate that the binding between colipase and lipase in the presence of substrate is strong and occurs in an approximately stoichiometric relationship.     

Bile salts and calcium absorption

1. The study of the effect of bile salts on enhancing calcium absorption in the rachitic chick has been extended to bile salts not present in chick bile, e.g. glycine conjugates and bile alcohol sulphates. 2. Bile and bile salts cause an increase in calcium absorption from sparingly soluble calcium hydrogen phosphate when compared with a suspension of calcium hydrogen phosphate in saline. 3. If the bile ducts of normal rats are tied the absorption of calcium from calcium hydrogen phosphate decreases but can be restored by giving bile salts with the calcium salt. 4. Bile salts increase solubility in water of the sparingly soluble calcium salts, phytate and phosphate at pH values between 6 and 8. 5. Bile salts increase the solubility in lipid solvents of calcium in approximately the same proportion as they increase the absorption of calcium from the gut. 6. The physiological role of bile in calcium absorption and its mode of action are discussed.     

Relationships between bile salts hydrophilicity and phospholipid composition in bile of various animal species

Bile salts and phospholipids from bile of chicken, dog, sheep, rat, ox, pig, guinea-pig and man were analyzed by high-performance liquid chromatography. Bile salts showed marked differences in their hydrophilic properties, owing to hydroxyl structure and type of conjugation. Phospholipids were generally similar, containing 90-95% of phosphatidylcholine which was made of molecular species containing palmitic acid in the sn-1 position. The comparative analysis of bile salts and phosphatidylcholines profile demonstrated that bile salts hydrophilicity influences the quantity of phosphatidylcholine in bile but not the quality.     

Response of enteric luminous bacteria to environmental conditions in the gut of the fish

The response of luminous bacterial cultures to conditions encountered in the fish gut such as neutral pH, the presence of bile salts, gastric juice and lysozyme was examined. The organisms preferred neutral pH. Bile salts did not inhibit their growth. Neither lysozyme nor gastric juice affected their growth and viability to any extent. In the light of these findings, the adaptability of luminous bacteria to conditions existing in the gut of fish was discussed.     

Response of enteric luminous bacteria to environmental conditions in the gut of the fish

The response of luminous bacterial cultures to conditions encountered in the fish gut such as neutral pH, the presence of bile salts, gastric juice and lysozyme was examined. The organisms preferred neutral pH. Bile salts did not inhibit their growth. Neither lysozyme nor gastric juice affected their growth and viability to any extent. In the light of these findings, the adaptability of luminous bacteria to conditions existing in the gut of fish was discussed.     

Effect of Acidity on the Composition of an Indigenous Soil Population of Rhizobium trifolii Found in Nodules of Trifolium subterraneum L.

Acidity affected which members of an indigenous soil population of Rhizobium trifolii nodulated Trifolium subterraneum L. cv. Mt. Barker. In three experiments involving plants grown either in mineral salts agar adjusted to pH 4.8 or 6.8 and inoculated with a soil suspension or grown directly in samples of unamended soil (pH 4.8) or soil amended with CaCO₃ (pH 6.4), 121 of 151 isolates of R. trifolii were placed into four serogroups. Seventy-nine of these isolates were placed into two serogroups (6 and 36) whose nodulating ability was affected by the pH of the plant root environment. Representatives of serogroup 6 occupied the greatest percentage of the nodules at the low pH in both mineral salts agar (77%) and in unlimed soil (47 and 57%). The same serogroup was a minor nodule occupant at the higher pH in mineral salts agar (0%) and in limed soil (0 and 10%). In contrast, serogroup 36 was virtually absent in nodules formed at the low pH, whereas it was the dominant serogroup at the higher pH in both mineral salts agar (32%) and in limed soil (35 and 49%). Despite the isolates from within each serogroup being antigenically identical, separation of cellular proteins by sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis revealed four and six different gel types within serogroups 6 and 36, respectively. Isolates represented by one or two gel types dominated the contribution of each serogroup to the nodule population. Further evidence for differences between isolates within each gel type were revealed from measurements of symbiotic effectiveness.     

Complete resistance to challenges with Hymenolepis nana cysticercoids derived from mouse, rat and beetle in mice

and 1986. Complete resistance to challenges with Hymenolepis nana cysticercoids derived from mouse, rat and beetle in mice. International Journal for Parasitology 16: 623–628. When BALB/c and dd strains of mice were given eggs of Hymenolepis nana, they all became completely resistant not only to challenge with mouse-derived cysticercoids but also to challenges with rat-derived and beetle-derived cysticercoids. Serum IgG antibodies at 47–60 days post egg inoculation reacted strongly with these three different host-derived cysticercoids when examined by IFA test, but IgA and IgM isotypes reacted very weakly. Antibodies of infected mouse sera (IgG, IgM and IgA were examined) reacted not only with the protoscolex (scolex of the excysted juvenile) but also with the outer cyst wall. By contrast, uninfected mouse sera and immune sera prepared seven days post cysticercoid inoculation did not react at all. Antigens of both cyst wall and protoscolex appeared to be of parasite origin and not of host origin, and appeared similar in parasites from the different host species.     

Isolation and identification of bile salts conjugated with cysteinolic acid from bile of the red seabream, Pagrosomus major.

Bile salts present in gallbladder of wild and cultured red seabream, Pagrosomus major, a marine teleost were analyzed. The bile from wild red seabream was found to contain two previously unknown bile salts along with two known bile salts, taurocholate and taurochenodeoxycholate. Isolation of each bile salt was performed by column chromatography. Fast atom bombardment mass spectra of the unknown bile salts showed the molecular ions (M-H)- of m/z 544 and 528 which are shifted 30 mass units upfield compared to those (m/z 514 and 498) of taurocholate and taurochendeoxycholate, respectively; this is consistent with the presence of cysteinolic acid (mol wt 155) instead of taurine (mol wt 125). Enzymatic hydrolysis of the bile salts released cholic acid and chenodeoxycholic acid, respectively, and an amino acid that was identified as D-cysteinolic acid by direct comparison with an authentic sample. From these results, the bile salts in the bile of wild red seabream were identified as the conjugates of cholic acid and chenodeoxycholic acid with cysteinolic acid. 1H- and 13C-magnetic resonance spectra of the bile salts were also consistent with the proposed structure. The cysteinolic acid conjugates were found only in wild and not in cultured red seabream; this distinction seems to result from differences in dietary cysteinolic acid.     

Effect of Acidity on the Composition of an Indigenous Soil Population of Rhizobium trifolii Found in Nodules of Trifolium subterraneum L

Acidity affected which members of an indigenous soil population of Rhizobium trifolii nodulated Trifolium subterraneum L. cv. Mt. Barker. In three experiments involving plants grown either in mineral salts agar adjusted to pH 4.8 or 6.8 and inoculated with a soil suspension or grown directly in samples of unamended soil (pH 4.8) or soil amended with CaCO(3) (pH 6.4), 121 of 151 isolates of R. trifolii were placed into four serogroups. Seventy-nine of these isolates were placed into two serogroups (6 and 36) whose nodulating ability was affected by the pH of the plant root environment. Representatives of serogroup 6 occupied the greatest percentage of the nodules at the low pH in both mineral salts agar (77%) and in unlimed soil (47 and 57%). The same serogroup was a minor nodule occupant at the higher pH in mineral salts agar (0%) and in limed soil (0 and 10%). In contrast, serogroup 36 was virtually absent in nodules formed at the low pH, whereas it was the dominant serogroup at the higher pH in both mineral salts agar (32%) and in limed soil (35 and 49%). Despite the isolates from within each serogroup being antigenically identical, separation of cellular proteins by sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis revealed four and six different gel types within serogroups 6 and 36, respectively. Isolates represented by one or two gel types dominated the contribution of each serogroup to the nodule population. Further evidence for differences between isolates within each gel type were revealed from measurements of symbiotic effectiveness.     

Taenia saginata: Production and characterization of monoclonal antibodies against Taenia saginata metacestode antigens

Cysticercosis is a major cause of economic loss in bovine production due to meat condemnation. Chemotherapy is being used in Brazilian cattle and a diagnostic test to improve the treatment program is desired. We produced monoclonal antibodies against crude (TAEB) and cyst fluid (TAEF) Taenia saginata metacestode antigens using immunized BALB/c mice. After cell fusion, 10 TAEB and nine TAEF hybrids were selected and cloned resulting in 18 IgG₁ and 32 IgM TAEB clones, and 9 IgG₁ and 9 IgM TAEF clones. Ascites was produced and Western blot testing was performed resulting in reactivity to protein fractions of low molecular weight (<18 kDa), 43, 55, 66 and 100 kDa. The indirect immunofluorescence test, with one monoclonal antibody against crude and one against cyst fluid antigens, recognized antigenic fractions of both the scolex and the bladder wall of metacestodes from naturally infected bovine.     

Ultrastructural studies of the cysticercoid of moniezia expansa (Anoplocephalidae) with special reference to the development of the cyst.

Cysticercoids of the sheep tapeworm Moniezia expansa have been grown in oribatid mites in the laboratory. Three species of mite became infected: Xenillus tegeocranus, Platynothrus peltifer and Euzetes globulus, the latter not previously recorded as a host of anoplocephalines. Cysticercoids aged 15 and 28 weeks were examined with the electron microscope. Four distinct types of cells were found in the 15-week cysticercoid. Subsequent cyst development involves a transformation from a cellular to a mainly fibrous structure. The fibres, arranged in three layers, resemble collagen fibres described elsewhere. The outer epidermis of the cyst is replaced by an amorphous, electron-dense outer coat whose nature is unknown. The inner part of the cyst becomes condensed to a myelin-like structure. The scolex develops features characteristic of the adult tapeworm.     

Pyrophosphate is a source of phosphoryl groups for Escherichia coli protein phosphorylation

Abstract Stress tolerance and cross-protection in Enterococcus faecalis ATCC19433 were examined after exposure to bile salts, acid or heat shock. Bile salts and heat adapted cells demonstrated induced homologous tolerance and cross-resistance. No cross-protection of heat adapted cells against acid stress is observed and pretreatment with bile salts even sensitized the cells to this challenge. Whole-cell protein extract analysis revealed that each treatment induced a battery of stress proteins. Some of these polypeptides are induced by more than one treatment. The greatest overlap is observed between bile salts and heat treatments. Eighteen stress proteins, including DnaK and GroEL, are common between these stresses.     

Cloud-point temperatures for lysozyme in electrolyte solutions: effect of salt type, salt concentration and pH

Liquid-liquid phase-separation data were obtained for aqueous saline solutions of hen egg-white lysozyme at a fixed protein concentration (87 g/l). The cloud-point temperature (CPT) was measured as a function of salt type and salt concentration to 3 M, at pH 4.0 and 7.0. Salts used included those from mono and divalent cations and anions. For the monovalent cations studied, as salt concentration increases, the CPT increases. For divalent cations, as salt concentration rises, a maximum in the CPT is observed and attributed to ion binding to the protein surface and subsequent water structuring. Trends for sulfate salts were dramatically different from those for other salts because sulfate ion is strongly hydrated and excluded from the lysozyme surface. For anions at fixed salt concentration, the CPT decreases with rising anion kosmotropic character. Comparison of CPTs for pH 4.0 and 7.0 revealed two trends. At low ionic strength for a given salt, differences in CPT can be explained in terms of repulsive electrostatic interactions between protein molecules, while at higher ionic strength, differences can be attributed to hydration forces. A model is proposed for the correlation and prediction of the CPT as a function of salt type and salt concentration. NaCl was chosen as a reference salt, and CPT deviations from that of NaCl were attributed to hydration forces. The Random Phase Approximation, in conjunction with a square-well potential, was used to calculate the strength of protein-protein interactions as a function of solution conditions for all salts studied.     

Microthrix pattern of Pseudogilquinia thomasi (Palm, 2000) (Cestoda: Trypanorhyncha) and a review of surface ultrastructure within the family Lacistorhynchidae Guiart, 1927

The microthrix pattern of larvae of Pseudogilquinia thomasi (Palm, 2000) is described for the first time using scanning electron microscopy. The surface ultrastructure of this species consists of three main forms of microtriches: papilliform filitriches, acicular filitriches and quadridigitate to octadigitate palmate spinitriches. The bothria are covered with palmate spinitriches interspersed on some parts with papilliform filitriches. Palmate spinitriches with papilliform, acicular and capilliform filitriches adorn the pars vaginalis and at the anterior part of the pars bulbosa, there is a transition zone in which the palmate spinitriches are replaced by papilliform to acicular filitriches towards the end of the scolex. ANOVA tests with Duncan's post hoc analyses revealed that there are significant differences in the length of spinitriches and their prongs, whether on the surfaces of the bothria or on the surfaces of the scolex peduncle. Callitetrarhynchus gracilis Pintner, 1931 is the second lacistorhynchid species for which surfaces of the segments were examined and the occurrence of large structures called scutes are reported for the first time from this species. There were also significant differences in the base length as well as height of the scutes in different parts of the strobila. In addition to describing the surface ultrastructures of the two lacistorhynchid species, all the available information on the microtriches within the family Lacistorhynchidae is reviewed herein and generic diagnoses related to microtriches are amended. J. Morphol. 277:394–404, 2016. © 2015 Wiley Periodicals, Inc.     

Role of electrostatic forces in hydroxy and keto bile salt-albumin interactions: some experimental observations.

Proton binding to bovine serum albumin and effects on hydroxy and keto bile salts-albumin binding were studied within a pH range between 5 and 10. Electrostatic forces contribute to the binding of these ligands to albumin; prototropic groups of albumin such as imidazol are involved in the interaction. Bile salts binding produces a shift in pK of these groups. It is postulated that hydroxy bile salt-albumin binding is linked with the N in equilibrium with B transition of the protein, while for keto bile salts a microarrangement in the protein binding sites is driving the interaction.     

Histopathological and ultrastructural studies of the tapeworm Monobothrium wageneri (Caryophyllidea) in the intestinal tract of tench Tinca tinca

Monobothrium wageneri is a monozoic caryophyllidean tapeworm of tench Tinca tinca. The pathological changes caused by this parasite within the intestinal tract of wild tench are described for the first time. Parasites were found attached to the anterior third of the intestine in tight clusters comprising up to 109 tapeworms. Infection was associated with the formation of raised inflammatory swellings surrounding the parasites. This host response, combined with the deep penetration of the scolex into the gut wall, formed a very firm seat of parasite attachment. Histopathological changes were characterised by a pronounced fibrogranulomatous lesion that extended through all layers of the intestine. This was accompanied by haemorrhage, oedema, necrosis and degeneration of the muscularis. A marked eosinophilic interface layer between the scolex of the tapeworm and gut wall indicated intimate host-parasite contact. Ultrastructural examinations revealed coniform spinitriches covering the neck and lateral sides of the scolex and capilliform filitriches present on the apical end of the scolex. Numerous glandular cytons (tegumental glands) were recorded throughout the scolex tegument. Large numbers of secretory granules discharged from the glands through a network of processes onto the scolex surface were consistent with distancing the cellular responses of the host. Observations of severe inflammatory lesions, partial intestinal occlusion and the potential for intestinal perforation represent important pathological changes that are consistent with loss of normal gut function. The lesions associated with the attachment of M. wageneri are more severe than those recorded for any other tapeworm of British freshwater fish.     

Bile and the absorption of strontium and iron

1. Strontium absorption was studied in vivo with loops of ileum in rachitic chicks and found to be increased by vitamin D(3), bile salts and sodium lauryl sulphate. 2. Bile salts and sodium lauryl sulphate rendered strontium soluble in butanol-benzene (1:1, v/v). 3. Bile was not concerned in the absorption of iron in rats from its water-soluble form, ferrous sulphate. 4. Ligation of the bile ducts in rats caused a decrease in the absorption of iron presented as its sparingly soluble phosphate. 5. The effect of bile on cation absorption is discussed.     

ANOMALIES IN THE ABSORPTION SPECTRUM AND BLEACHING KINETICS OF VISUAL PURPLE

Visual purple from winter frogs shows an intermediate yellow color during bleaching by light; summer extractions do not. This seasonal effect can be duplicated by variations in the hydrogen ion concentration and in the temperature of the solutions. Increasing the pH approximates the summer condition, while decreasing the pH approximates the winter condition. Temperature has no effect on the bleaching of alkaline solutions but greatly influences acid solutions. At low temperatures the bleaching of add solutions resembles the winter condition, while at higher temperatures it resembles the summer condition. A photic decomposition product of frog retinal extractions is an acid-base indicator: it is yellow in acid and colorless in alkaline solution. Its color is not dependent upon light. The hydrogen ion concentration of visual purple solutions does not change under illumination, nor is there a difference in the pH of summer and winter extractions. Bile salt extractions of visual purple are usually slightly acid. The conflicting results of past workers regarding the appearance of "visual yellow" may be due to seasonal variation with its differences in temperature, or to the presence of base in the extractions. It is also possible that vitamin A may be a factor in the seasonal variation. The photic decomposition of visual purple in bile salts solution, extracted from summer frogs, follows the kinetics of a first order reaction. Visual purple from winter frogs does not conform to first order kinetics. Photic decomposition of alkaline, winter visual purple extractions also follows a first order equation. Acid, winter extractions appear to conform to a second order equation, but this is probably an artefact due to interference by the intermediate yellow.