Identification and typing of food-borne Staphylococcus aureus by PCR-based techniques

The possibility of using PCR for rapid identification of food-borne Staphylococcus aureus isolates was evaluated as an alternative to the API-Staph system. A total of 158 strains, 15 S. aureus, 12 other staphylococcal species, and 131 isolates recovered from 164 food samples were studied. They were phenotypically characterized by API-Staph profiles and tested for PCR amplification with specific primers directed to thermonuclease (nuc) and enterotoxin (sea to see) genes. Disagreement between the PCR results and API-Staph identification was further assessed by the analysis of randomly amplified polymorphic DNA (RAPD) profiles obtained with three universal primers (M13, T3, and T7) and 16S rDNA sequencing. Forty out of 131 isolates (31%) tested positive for PCR enterotoxin. Of these, 14 (11%) were positive for sea, 22 (17%) for sec, one (0.8%) for sed, and three (2.2%) for sea and sec. No amplification corresponding to seb nor see was obtained. Cluster analysis based on RAPD profiles revealed that most of the sec positive food isolates grouped together in three clusters. Cluster analysis combining the three RAPD fingerprints (M 13, T3, and T7), PCR-enterotoxin genotype and API-Staph profiles, grouped the nuc PCR positive isolates together with S. aureus reference strains and the nuc PCR negative isolates with reference strains of other staphylococcal species. The only nuc PCR positive food isolate that remained unclustered was a sed positive strain identified by 16S rDNA sequence as S. simulans. The high concordance between S. aureus and nuc PCR positive strains (99%) corroborates the specificity of the primers used and the suitability of nuc PCR for rapid identification of S. aureus in routine food analysis.     

洞庭湖外睾吸虫新种及其生活史

本文报告洞庭湖区鲶鱼肠道寄生的洞庭湖外睾吸虫Exorchis dongtinghuensis sp.nov(新种)及其全程生活史,其第一中间宿主为湖北钉螺Oncomelania hupensis;第二中间宿主为鲤鱼、鲫鱼和金鱼;终宿主为鲶鱼Parasilurus asotus。作者对各期宿主作了人工感染试验和现场自然感染调查。对其发育过程作了观察比较。     

Potamopyrgus antipodarum(Mollusca:Hydrobiidae) in continental aquatic gastropod communities: impact of salinity and trematode parasitism

The structure of gastropod communities was examined from January to June 1999 in four sites of the streams of Mont Saint-Michel Bay along a gradient of salinity, and the occurrence of larval trematodes infecting snails was studied. Abundance and species richness of gastropods increased from polyhaline (95 snails, 1 species) to oligohaline waters (6672 snails, 6 species). Whatever the salinity, the most abundant species was Potamopyrgus antipodarum, an invasive non-indigenous species that represented 80% of the gastropods. Only one male was found in P. antipodarum populations suggesting a predominantly parthenogenetic mode of reproduction. Among 7218 gastropods collected, 1.2% were infected by larval trematodes: 5 species in Lymnaea peregra (4.4%), 4 species in Planorbis planorbis (12.0%), one echinostome in Physa acuta (0.2%), and a new species of Sanguinicola in P. antipodarum (0.5%). This is the first record of infected P. antipodarum in Europe. No parasites were found in polyhaline waters. The prevalence per host population varied from 0 to 100% depending on time of collection, salinity and host species. In the lowest-salinity site, abundance of gastropods and prevalence of trematodes were negatively correlated. The dominance of P. antipodarum in the gastropod communities is discussed in relation with euryhalinity, parthenogenesis and weak rate of parasitism.     

Four trematode cercariae from the New Zealand intertidal snail Zeacumantus subcarinatus (Batillariidae)

Abstract

The cercariae and sporocysts (or rediae) of four trematode species are described from the intertidal snail Zeacumantus subcarinatus: a distome xiphidiocercaria assigned to the genus Renicola (family Renicolidae); a monostome xiphidiocercaria belonging either to the genus Microphallus or Megalophallus (family Microphallidae); a magnacercous cercaria of the genus Galactosomum (family Hetero‐phyidae); and a cercaria of the genus Philophthalmus (family Philophthalmidae). The morphological features of these cercariae are compared to previously described cercariae of the same genera. In addition, since the philophthalmid cercaria encysts readily on artificial substrates in the laboratory, the metacercaria of this species is also described. These cercariae are part of a diverse community of at least six digenean species parasitising the snail Z. subcarinatus that, together, have a major impact on the ecology and evolution of this snail.     

The host antigen phenomenon in experimental murine schistosomiasis: The transfer of 3-week old Schistosoma mansoni between two inbred strains of mice

A surgical technique for the transfer of 3-week old Schistosoma mansoni to the mesenteric veins of mice is described. Parasites grown in a donor C3H/StCrl strain survive on transfer to recipient C57BL/10J mice despite prior immunization with C3H/StCrl cells or skin grafts. The findings indicate that the ‘species-specific’ mouse host antigens associated with adult worms in xenogeneic transfers do not have an allogeneic basis in these two strains.     

Changes of phospholipid fatty acid composition in the digestive gland of the mollusc <Emphasis Type="Italic">Littorina saxatilis</Emphasis>, caused by trematode larvae

Lipids in the digestive gland of the mollusc Littorina saxatilis from the White and Barents Seas were studied. Changes of their biochemical composition are discussed in the connection with different temperature of the habitat and with infestation with trematode larvae. Comparative analysis of the fatty acid (FA) composition of each of the phospholipids in intact molluscs has revealed essential differences. Phosphatidylcholine and monophosphatidylinositol (MPI) FA did not differ in the ω3/ω6 ratio, which is due to their tolerance to the temperature factor, whereas more unsaturated phospholipids—phosphatidylethanolamine (PEA), its plasmalogen form (pPEA), and phosphatidylserine—differed 1.5–2 times in the studied molluscs. Prevalence of ω3-acids in Barents Sea mollusc is associated undoubtedly with lower habitat temperatures, as this provides a higher fluidity of the membrane phospholipids. Infestation affected to the greatest degree the quantitative FA composition in pPEA and MPI. At infestation, there was revealed a threefold decrease of the content of eicosenoic FA, C20:1, only in MPI among all considered phospholipids, whereas it was increased, in the contrary, in all other phospholipids. Monophosphatidylinositols differed essentially from other phospholipids by the saturated FA amount as well, which changed the unsaturation index of these phospholipids. Since the functional significance of this minor phospholipid is determined by its participation in the so-called phosphatidylinositol system of the hormonal signal transduction, it seems interesting to elucidate whether the increase of this membrane phospholipid saturation at invasion affects the reflex connection between signals from receptors located in the parasite and enzymatic processes.     

Haplorchis pumilio and H. taichui in Vietnam discriminated using ITS-2 DNA sequence data from adults and larvae

Samples of Haplorchis taichui and Haplorchis pumilio of different life-stages (cercariae, metacercariae and adults) and from different host species (snail, fish, dog, cat and human) were collected in Nghe An and Nam Dinh Provinces in Vietnam. Samples from Thailand were available for comparison. All adults and metacercariae were initially identified using morphological criteria. Polymerase chain reaction (PCR) assays were developed for discriminating between the species. The complete sequence for the nuclear ribosomal internal transcribed spacer-2 (ITS-2) was obtained from one adult and one metacercaria of H. taichui and three adults and three metacercariae of H. pumilio from Vietnam. Sequences from cercariae from three different snails clustered with those of H. pumilio. Intra-individual variation in the ITS-2 region was detected by sequencing of cloned PCR products. These are the first sequences from Vietnamese Haplorchis spp. to be reported and demonstrate that H. taichui and H. pumilio can be identified unambiguously from any life-stage, including the cercarial stage that is difficult to identify using morphology. Discrepancies in the literature are discussed and examples of apparent misidentification highlighted. The data provide a resource to assist in taxonomic studies on heterophyids, in the design of probes for diagnosis and for field surveys to identify infection in snails.     

Comparative ultrastructural topography of the gut epithelia of some trematodes

The gut epithelia of six species of digenetic trematodes, Clonorchis sinensis, Eurytrema pancreaticum, Haematoloechus lobatus, Echinostoma hortense, Schistosoma japonicum and Fasciola hepatica, were studied with scanning and also transmission electron microscopy. Morphological differences in cytoplasmic projections of the gut of adult flukes were demonstrated stereoscopically among these species. The cytoplasmic projections vary considerably in shape, but are roughly separated into three groups by their essential forms: ribbon-shaped narrow type in C. sinensis and E. pancreaticum, broad, triangular with filamentous extensions distally and/or marginally as in F. hepatica and E. hortense, and broad, sheet-like or triangular with the distal ends blunt or rounded as in H. lobatus and S. japonicum. This character appears rather constant, without regional differences in the gut. No marked correlation was found between the gut projections of the parasites and their host or food. There are also specific discriminations in the ultrastructure of the cellular organization among the species examined.     

Conspicuous behaviour of Fundulus heteroclitus associated with high digenean metacercariae gill abundances

Fundulus heteroclitus from six sites throughout the Hackensack Meadowlands District in northern NJ, U.S.A., were examined. Differences in behaviour (surfacing, conspicuousness and activity) were compared to gill infection intensity. Fish from populations infected with >1500 digenean metacercariae of Ascocotyle phagicola diminuta and Echinochasmus schwartzi , spent significantly more time at the water surface and exhibited significantly more conspicuous behaviour ( e.g. jerking) than fish from less parasitized populations. This more conspicuous behaviour has the potential to increase trophic transmission of the parasite to its definitive wading-bird host.     

Proving the antimicrobial spectrum of an amphoteric surfactant-sol-gel coating: a food-borne pathogen study

An antimicrobial coating was evaluated in this work for its antimicrobial efficacy against common food-borne pathogens. Dodecyl-di(aminoethyl)-glycine, an organic disinfectant, was immobilized in a silicon oxide matrix to generate thin films over surfaces by means of the sol-gel process. Tetraethoxysilane was used as the polymeric precursor. No alteration of optical transparency on the covered surfaces was observed. Topographic images obtained with atomic force microscopy showed a homogeneous film with no additional roughness added by the polymer to the surface. The attenuated total reflectance-Fourier transform infrared spectral data showed the presence of dodecyl-di(aminoethyl)-glycine in the silicon oxide network after a normal cleaning procedure. The antimicrobial efficacy test was performed by exposing coated slides to suspensions of common food-borne pathogens: Escherichia coli, Staphyloccocus aureus, E. coli O157:H7, Salmonella typhi, S. cholerasuiss, Listeria innocua and L. monocytogenes. The coating activity was not only bacteriostatic but also bactericidal. The percent reduction of viable microorganism exposure over 24 h to the coated surface ranged between 99.5%, for the more resistant gram-positive bacteria, and over 99.999%, for most gram-negative bacteria. The silicon matrix itself did not account for any reduction of viable microbial, even more an increase was observed.