The soil flagellate Heteromita globosa accelerates bacterial degradation of alkylbenzenes through grazing and acetate excretion in batch culture.

The impact of grazing by soil flagellates Heteromita globosa on aerobic biodegradation of benzene by Pseudomonas strain PS+ was examined in batch culture. Growth of H. globosa on these bacteria obeyed Monod kinetics (mu(max), 0.17 +/- 0.03 h(-1); K(s), 1.1 +/- 0.2 x 10(7) bacteria mL(-1)) and was optimal at a bacteria/ flagellate ratio of 2000. Carbon mass balance showed that 5.2% of total [ring-U-(14)C]benzene fed to bacteria was subsequently incorporated into flagellate biomass. Growth-inhibiting concentrations (IC50) of alkylbenzenes (benzene, toluene, ethylbenzene) were inversely related with their octanol/ water partitioning coefficients, and benzene was least toxic for bacteria and flagellates with IC50 values of 4392 (+/- 167) microM and 2770 (+/- 653) microM, respectively. The first-order rate constant for benzene degradation (k1, 0.48 +/- 0.12 day(-1)) was unaffected by the presence or absence of flagellates in cultures. However, the rate of benzene degradation by individual bacteria averaged three times higher in the presence of flagellates (0.73 +/- 0.13 fmol cell(-1) h(-1)) than in their absence (0.26 +/- 0.03 fmol cell(-1) h(-1)). Benzene degradation also coincided with higher levels of dissolved oxygen and a higher rate of nitrate reduction in the presence of flagellates (p < 0.02). Grazing by flagellates may have increased the availability of dissolved oxygen to a smaller surviving population of bacteria engaged in the aerobic reactions initiating benzene degradation. In addition, flagellates may also have increased the rate of nitrate reduction through the excretion of acetate as an additional electron donor for these bacteria. Indeed, acetate was shown to progressively accumulate in cultures where flagellates grazed on heat-killed bacteria. This study provided evidence that grazing flagellates stimulate bacterial degradation of alkylbenzenes and provide a link for carbon cycling to consumers at higher trophic levels. This may have important implications for bioremediation processes.

The predatory soil flagellate Heteromita globosa stimulates toluene biodegradation by a Pseudomonas sp.

A model food chain was established to investigate the influence of grazing by flagellates on bacteria degrading toluene in batch culture. The rate of toluene consumed by a Pseudomonas sp. strain PS+ (max. 0.37 fmol cell(-1) h(-1)) was significantly higher in the presence of the bacterivorous flagellate Heteromita globosa (max. 1.38 fmol cell(-1) h(-1)). A maximum increase of up to 7.5 times was observed in the rate of toluene consumed by these bacteria during exponential growth of this flagellate. Carbon conversion efficiency (CCE) of bacteria to flagellate biomass was estimated to be 33.4% based on measured biovolumes and published values for carbon contents. However, the CCE for toluene-derived carbon was lower (max. 4.9%) when calculations were based on incorporation of [ring-U-(14)C]toluene into biomass of flagellates grazing on labelled bacteria. The findings suggest a potential role for flagellates in bioremediation processes.

Ultrastructure and histochemistry of the tegument of juvenile paramphistomes during migration in Indian ruminants.

The tegument of juvenile Paramphistomum epiclitum and Fischoederius elongatus (Paramphistomidae: Digenea) resembles those of other digeneans. Seven types of papillae were observed, mostly on the oral and acetabular surfaces, and increase in number during migration. Also evident are two types of secretory body (T1 and T2) which are synthesized separately in tegumental cytons underlying the syncytium. Exocytosis of T2 bodies occurs at the apical membrane and appears to contribute to a fibrous glycocalyx. The tegumental syncytium lining the pharynx and acetabulum is thinner and has a higher capacity for vacuolation than the general tegument. These may represent important sites for osmoregulation. The absence of mitochondria from the tegument in migrating juveniles suggests limited involvement in energy demanding processes. Pigmentation of the subtegument is first evident in mature cercariae and is progressively eliminated during migration.

Ultrastructure and histochemistry of the digestive tract of juvenile Paramphistomum epiclitum (Paramphistomidae: Digenea) during migration in Indian ruminants.

The digestive tract of juvenile Paramphistomum epiclitum consists of a foregut with a highly muscular terminal pharynx and an oesophagus, which leads to a pair of unbranched and blind-ending intestinal caeca. A syncytium lining the foregut is continuous with the external tegument and displays similar sensory papillae and secretory bodies (T1 and T2). A third type of secretory body (T3) is confined to the oesophageal cytons of newly excysted juveniles and is first evident in the syncytium by day 14 of migration. An epithelium lining the caeca is composed of a single layer of morphologically uniform cells whose apical surface is amplified by microvilli. Dense secretions synthesized in the caecal epithelium of mature cercariae are released during migration by a mechanism resembling modified apocrine discharge. The caecal epithelium of migrating juveniles undergoes a 10-fold increase in surface amplification (irrespective of growth) during its transition from a primarily secretory tissue to one apparently specialized for absorption.

Ultrastructure and histochemistry of the protonephridial system of juvenile Paramphistomum epiclitum and Fischoederius elongatus (Paramphistomidae: Digenea) during migration in Indian ruminants.

The protonephridial system of juvenile Paramphistomum epiclitum and Fischoederius elongatus consists of a bilaterally symmetrical arrangement of primary, secondary and tertiary ducts which connect individual flame cells with a simple common bladder. Primary and secondary ducts are formed from columns of adjoining cells which provide an epithelial lining, whose luminal surface is elaborated with either short tubercles or lamellae. Groups of cilia project from the luminal surface at frequent intervals along secondary ducts. By contrast, the tertiary ducts and bladder are lined with a nucleated syncytium which ends at a junctional complex formed with the terminal canal. The latter is continuous with the tegumental syncytium and opens at a nephridiopore on the postero-dorsal surface. Tertiary ducts of mature cercariae contain concretions which are voided by migrating juveniles in whose tertiary ducts lipids are progressively accumulated. Evidence for the role of protonephridia in excretion and possibly in osmoregulation and ionic balance is currently examined.

Ultrastructure and histochemistry of the lymph system and parenchyma of juvenile paramphistomes (Paramphistomidae: Digenea) during migration in Indian ruminants.

The lymph system of juvenile Paramphistomum epiclitum and Fischoederius elongatus consists of a single pair of longitudinal primary vessels from which sub-dividing branches extend laterally to associate with most major tissues and organs. The system originates shortly after excystation in the definitive host and is fully developed in day 14 juveniles. Lymph vessels are syncytial and membrane limited, with a matrix which contains autophagic-like inclusions, clusters of SER and free nuclei. Similar organelles are evident in the matrix of parenchyma and specialized cells juxta-posing the pharynx (JP cells). These tissues are intimately associated and perhaps functionally integrated. Parenchyma represents a major site for carbohydrate storage and turnover, whilst the lymph appears to perform a similar role for proteins. The JP cells of juveniles display prolific autophagic-like activity only during migration, which coincides with the depletion of carbohydrate reserves in parenchyma. Key mitochondrial enzymes were histochemically demonstrated in the lymph despite the apparent absence of mitochondria from this system in post-day 14 juveniles. Succinate dehydrogenase activity was cytolocalized in mitochondria, whilst attempts to perform a similar localization of this enzyme in lymph were unsuccessful. The possibility of non-enzymatic interference in the histochemical demonstration of dehydrogenase is examined.