Evaluation of a monoclonal antibody-based immunoassay for detecting type B Clostridium botulinum toxin produced in pure culture and an inoculated model cured meat system

A monoclonal antibody-based amplified ELISA method for detecting Clostridium botulinum type B toxin was evaluated for its ability to detect the toxin in the supernatant fluid of pure cultures and after growth from Cl. botulinum spores inoculated into pork slurries. Slurries containing NaCl (1.5–4.5%w/v) and polyphosphate (0.3%w/v) were either unheated or heated 80°C/5 min followed by 70°C/2 h before incubation at 15°, 20° or 27°C. Presence of specific toxin was confirmed by mouse bioassay and results were compared with those of the amplified ELISA method. A total of 48 strains, consisting of 38 Cl. botulinum and 10 Cl. sporogenes (putrefactive anaerobes), and 140 slurry samples were tested. Cultures of eight out of nine strains of type B Cl. botulinum and 73 of 101 slurry samples containing type B toxin were positive by ELISA; the remaining 28 slurry samples contained type B toxin at levels below or close to the detection limit (20 LD₅₀/ml) of the type B ELISA. No falsepositive reactions occurred with Cl. botulinum types A, C, D, E or F, or with the 10 strains of Cl. sporogenes. Toxin produced by one strain of Cl. botulinum type B (NCTC 3807) was not detected by this single monoclonal antibody-based amplified ELISA. With a mixture of two monoclonal antibodies, however, the toxin from NCTC 3807 could be detected without reducing the sensitivity of the ELISA.     

Histological Aspects of Root Formation in Petioles of Detached Leaves of Pereskia grandifolia (Cactaceae): Natural Conditions and Effects of GA3 and Dark

Adventitious roots arise naturally on petioles of Pereskia grandifoliaHaw. held in light. At about the 12th day after the beginningof the experiment, the root primordia arise in a callus tissuedeveloped from the basal portion of the petiole. Associatedwith the development of the callus, noticeable structural changesoccur in the originating organ. Petioles maintained in darkalso form callus; however, they die in a few days. On the otherhand, petioles treated with GA,, maintained in light, developcallus and survive; but they do not give rise to roots. Someaspects are discussed, such as: the kind of origin observedfor the roots, and the possible physiological basis for theirformation, as wdl as for the inhibition of their appearance Pereskia grandifolia, adventitious root formation, gibberelljc acid, petiole structure, rooting     

Isolation and characterization of six microsatellites in the Mexican beaded lizard Heloderma horridum

Six polymorphic microsatellite loci were isolated and characterized in the threatened Mexican beaded lizard, Heloderma horridum. The number of alleles per locus ranged from three to 12, with observed heterozygosity estimates ranging from 0.00 to 0.77, and expected heterozygosity estimates ranging from 0.00 to 0.73. These microsatellites will provide a valuable tool for the investigation of the genetic variation and structure of both wild and captive H. horridum populations.     

Influence of an ecosystem engineer,the emergent macrophyte Sparganium erectum,on seed trapping in lowland rivers and consequences for landform colonisation

1. Plant physical ecosystem engineers can influence vegetation population and community dynamics by modifying, maintaining or creating habitats. They may also have the potential to act upon biotic processes, such as seed dispersal. 2. Examples exist of reduction in seed dispersal distances in vegetated compared to unvegetated terrestrial environments, and concentration of seed deposits associated with plant patches. Such effects in aquatic environments have been little studied, but the engineering effect of plant patches on patterns of flow velocity and sediment deposition in streams suggests that they may play a similar role. 3. In this study, we assess the potential of an emergent aquatic species, Sparganium erectum, to play a role in physically modifying river habitats and trapping seeds by examining patterns of seed deposition and substrate type in 47 river reaches across England and southern Scotland, U.K. 4. Areas of the river channel within or adjacent to S. erectum patches harboured more plant seeds and more species than unvegetated areas and had finer, sandier substrates with higher organic matter, total nitrogen and total phosphorus content. Most seed species were competitive, indicating that they were well suited to colonise the competitive environment of an S. erectum patch, and could potentially further stabilise accumulated sediments and contribute to landform development. 5. We demonstrate that S. erectum patches influence both the physical environment and the retention of seeds, in consistent patterns across the channel bed, for a range of lowland rivers that vary in stream power and geology and which can be expected to vary in levels of supply of fine sediment and seeds. 6. Our findings support the hypothesis that the fundamental influence of a riverine ecosystem‐engineering species on slowing fluid flow links the habitat creation process of sediment sorting and retention to seed trapping. We suggest the process is applicable to a wide range of aquatic and riparian vegetation. We also suggest that the mono‐specific and competitive growth, which is typical of these engineering species, will strongly influence the recruitment of trapped seeds.     

Coccidian Parasites of Intertidal Fishes from Wales: Systematics, Development, and Cytochemistry

SYNOPSIS. Examination of littoral fish Blennius pholis and Cottus bubalis caught at Aberystwyth and Porth Cwyfan, Wales. U.K., revealed 2 species of coccidia.
Eimeria dingleyi sp. n. Oocysts spherical (16.1–19.2) to subspherical (13.9–14.2 × 18.8–20.0) μm, with thin walls; sporulation outside the host to produce ellipsoid sporocysts; endogenous phases in epithelial cells throughout intestine; 26 of 58 B. pholis infected.
Eimeria variabilis (Thélohan) Reichenow. Oocysts spherical (11.9–14.6) to subspherical (9.2–10.9 × 13.9–14.3) μm; sporulation in lining of pyloric ceca and rectum; previously unrecorded schizonts and gametocytes present; 21 of 25 C. bubalis infected.
Electron microscopy revealed that the oocyst wall of E. variabilis consists of a thin membrane whereas the sporocyst wall is thick and 3-layered. Typical oocyst wall-forming bodies were absent from the macrogamete. Cytochemical tests on the endogenous stages of E. dingleyi and E. variabilis indicated that in general they resembled other coccidia in their chemical constitution.     

Use of conductance measurements to detect growth of Clostridium botulinum in a selective medium

Growth of Clostridium botulinum in a selective medium (SBM) was monitored using conductance measurements. The correlation of log ₁₀ counts of spores and vegetative cells with detection times was highly significant ( r = 0.96, 0.97; P < 0.001). Log₁₀ counts of Cl. botulinum growing in pasteurized pork slurries also showed a clear linear relationship with detection times ( r = 0.77) but the confidence limits (± log₁₀ 2.3) of the regression line were too wide to estimate numbers of Cl. botulinum. For growth studies of Cl. botulinum in pork slurries detection times were useful in determining whether or not a visually unspoiled sample contained growing cells. Knowledge that a sample contained growing cells allowed a count to be made within 24 h, whereas 48 h would elapse before results from a traditional plate count were available.