Stephen G. Zam

Some parasitic diseases of dolphins.

Lungworm infection (Halocercus sp.) in the Atlantic bottlenose dolphin (Tursiops truncatus) was found to be very common in animals captured from the wild and in those raised within an aquarium. Infection of the Amazon dolphin (Inia geoffrensis) with Hunterotrema caballeroi causes pulmonary atelectasis. An unidentified holotrich ciliate was found in the blowholes of over 50% of the animals examined and was responsible for a purulent pneumonia in 1 instance. Hepatic and pancreatic trematodiasis which caused interstitial fibrosis occurred frequently in T. truncatus; the trematode was identified as Campula palliata. Pholeter gastrophilus, a trematode, results in the proliferation of a tumorous mass of fibrous tissue in the wall of the stomach. The sclerotic tissue surrounds cavities containing parasites and ova.

Biogeochemical ecology of Thiothrix spp. In underwater limestone caves

Thiothrix spp., sulfide‐oxidizing mixotrophic bacteria, were sampled from visible colonies in the Floridan aquifer in several underwater caves, sinkholes, and springs below the water table in North Florida. Bacteria samples were collected by cave divers certified by the National Speleological Society/Cave Diving Section. Sites sampled were ecological niches in the aquifer where visible colonies had a white slimy or filamentous appearance indicative of Thiothrix spp. Sterile sampling methods were adapted to the underwater cave setting. Bulk water samples for media preparation were collected by divers from bacteria sampling sites. Bacteria were isolated and cultured in growth media prepared with cave or spring water. Thiothrix spp. were identified by microbiological and immunological methods. Monoclonal antibodies specific for Thiothrix spp. were utilized in fluorescent antibody assays and enzyme‐linked im‐munosorbent assays (ELISA). Thiothrix was found in six of eight underwater caves sampled. Three of the...

Detection of Salmonella enteritidis in environmental samples by monoclonal antibody-based ELISA.

We have developed a enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody (ASCII) for the detection of Salmonella enteritidis in environmental samples. ELISA was used to test for sensitivity and specificity of ASCII. 38 other species of bacteria, including 31 Salmonella species were included in cross-reactivity testing with ELISA. ASCII showed no reactivity with any other species tested. ASCII was found to be an IgG1 specific for S. enteritidis lipopolysaccharide (LPS). The lower limits for S. enteritidis detection was 10(5) cells/ml for pure cultures and in 10% sludge (w/v). Environmental samples (raw wastewater, wastewater effluents, mixed liquor and aerobically digested sludge) were obtained twice from five sites and ELISA tested for the presence of S. enteritidis. ELISA results compared to the American Public Health Association (APHA) method of Salmonella detection were not significantly different (P greater than 0.05). The ELISA took 24 h for completion compared to 96-120 h for the APHA procedure. Results demonstrate the reliability of the ELISA and, more importantly, provides a rapid means of detection of S. enteritidis in environmental samples.

Identification, enrichment, and isolation ofThiothrix spp. from environmental samples

We have developed a method to enrich, isolate, and identifyThiothrix spp. in environmental samples. This procedure employs low concentrations of organic compounds, the addition of reduced sulfur compounds (sulfide or thiosulfate), and preparation with spring water that containsThiothrix spp. The enrichment enhanced identification ofThiothrix spp. by promoting deposition of intracellular sulfur granules and inhibiting overgrowth by other bacteria. The relatively high calcium content of the spring water contributed to the culture procedure. With this technique,Thiothrix spp. were observed in two activated sludge systems, a municipal water storage tank, three springs, and four underground freshwater caves in the phreatic zone of the Floridan aquifer. Two differentThiothrix cultures have been isolated from a freshwater cave and a water storage tank by this procedure. It appears that media prepared with spring water known to supportThiothrix spp. can be designed to provide highly selective methods for isolation ofThiothrix spp. from a wide range of environments.

DETECTION AND QUANTIFICATION OF ORNITHODOROS-SPECIFIC ANTI-TICK ANTIBODY BY COMPETITIVE INHIBITION ELISA

The objective of this study was to develop a highly specific enzyme-linked immunosorbent assay (ELISA) for the serological detection of anti-Ornithodoros tick antibodies in animals. Affinity-purified rabbit anti-Ornithodoros IgG antibodies were employed in indirect competitive inhibition ELISA assays designed to measure the anti-Ornithodoros antibody titers in other animal species using the domestic goat (Capra hircus) as a large animal model. Repeated infestation of goats with Ornithodoros coriaceus was found to elicit the formation of antibodies capable of inhibiting the binding of the Ornithodoros-specific rabbit IgG. Western blot analysis of goat and rabbit anti-tick antisera demonstrated both animal species to respond immunologically to a set of 9 major protein bands in O. coriaceus salivary gland extracts. The results of these experiments demonstrate that a history of animal exposure to O. coriaceus may be detected serologically by competitive inhibition ELISA.

Immune recognition of ornithodoros tick (Acari: Argasidae) salivary antigens by anti-Psoroptes cuniculi antibodies

Rabbits infested with either Ornithodoros sp. ticks or Psoroptes cuniculi mites were assayed for anti-tick antibody by enzyme-linked immunosorbent assay. Titration of rabbit serum against Ornithodoros sp. salivary gland extract (SGE) demonstrated both mite- and tick-infested animals to have elevated anti-tick antibody titers. Western blot analysis demonstrated the anti-mite and anti-tick antisera to contain antibodies with affinities for both common and unique subsets of Ornithodoros SGE proteins.

Isolation and characterization of a non-specific esterase enzyme of Ascaris suum perienterc fluid☆

Abstract 1. 1. A non-specific esterase enzyme of Ascaris perienteric fluid was isolated with a tenfold increase in specific activity over the crude enzyme and a 6% recovery. 2. 2. The pH optimum of the non-specific esterases is pH 8·4, with a molecular weight greater than 300,000 and an apparent isoelectric point of pI 4·5−5·0. 3. 3. By its behavior to various inhibitors, the enzyme appears to be a B-type esterase. 4. 4. A lipase with a pH optimum of pH 7·4 hydrolyzing emulsified tributyrin was demonstrated.

Disc electrophoresis of Ascaris suum female reproductive system esterase enzymes

Abstract 1. 1. The isoenzyme pattern of soluble and membrane bound esterases, extracted with 1% Triton X-100, of Ascaris suum female reproductive tissues were identified by disc electrophoresis using various substrates and inhibitors. 2. 2. By their behavior to various inhibitors, all esterases of the reproductive tissues appear to be B-type esterases. 3. 3. No C-type esterase or lipase were demonstrated.