Salivary gland thrombostasin isoforms differentially regulate blood uptake of horn flies fed on control- and thrombostasin-vaccinated cattle.

Thrombostasin (TS) is an anticlotting protein found in saliva of Haematobia irritans (horn flies). The polymorphic nature of the ts gene was first associated with success of horn flies blood feeding on a laboratory host, New Zealand White rabbits. In this study, we report results of similar studies testing blood uptake of horn flies feeding on a natural host, cattle.

Chemical and electroporated transformation of Edwardsiella ictaluri using three different plasmids.

Transfer of DNA by conjugation has been the method generally used for genetic manipulation of Edwardsiella ictaluri because, previously, attempts to transform E. ictaluri by the uptake of naked DNA have apparently failed. We report here the successful transformation of seven strains of E.

Salivary gland thrombostasin isoforms differentially regulate blood uptake of horn flies fed on New Zealand White rabbits.

Thrombostasin (TS) is a previously characterized anticlotting protein with multiple isoforms found in the saliva of horn flies. In this report, the effect of TS isoforms on blood feeding was assessed using individual flies that carried corresponding ts allelles. Laboratory studies of horn fly blood feeding were conducted using colony-reared flies fed on New Zealand White (NZW) rabbits.

In vitro and in vivo interaction of macrophages from vaccinated and non-vaccinated channel catfish (Ictalurus punctatus) to Edwardsiella ictaluri.

Macrophages from catfish vaccinated with an Edwardsiella ictaluri vaccine and macrophages from non-vaccinated catfish were used in in vitro and in vivo studies with red-fluorescent E. ictaluri to assess phagocytic ability, reactive oxygen and nitric oxide production and bactericidal activity. In the in vitro experiment, macrophages were harvested from vaccinated and non-vaccinated fish and then exposed to red-fluorescent E.

Multiplex-PCR for simultaneous detection of 3 bacterial fish pathogens, Flavobacterium columnare, Edwardsiella ictaluri, and Aeromonas hydrophila.

A multiplex PCR (m-PCR) method was developed for simultaneous detection of 3 important fish pathogens in warm water aquaculture. The m-PCR to amplify target DNA fragments from Flavobacterium columnare (504 bp), Edwardsiella ictaluri (407 bp) and Aeromonas hydrophila (209 bp) was optimized by adjustment of reaction buffers and a touchdown protocol. The lower detection limit for each of the 3 bacteria was 20 pg of nucleic acid template from each bacteria per m-PCR reaction mixture.

Induced cross-protection in channel catfish, Ictalurus punctatus (Rafinesque), against different immobilization serotypes of Ichthyophthirius multifiliis.

Abstract Channel catfish, Ictalurus punctatus (Rafinesque), were immunized with Ichthyophthirius multifiliis (Ich) theronts and trophonts, and the immune response and host protection against both homologous and heterologous serotypes of Ich were evaluated. Immunizations were done with two immobilization serotypes (ARS4 and ARS6) of live theronts by bath immersion (trial I) and with sonicated trophonts by intraperitoneal (i.p.

Analysis of 16S-23S intergenic spacer regions of the rRNA operons in Edwardsiella ictaluri and Edwardsiella tarda isolates from fish.

Aims: To analyse interspecies and intraspecies differences based on the 16S-23S rRNA intergenic spacer region (ISR) sequences of the fish pathogens Edwardsiella ictaluri and Edwardsiella tarda.

Methods And Results: The 16S-23S rRNA spacer regions of 19 Edw. ictaluri and four Edw.

Antigenicity of Streptococcus agalactiae extracellular products and vaccine efficacy.

Streptococcus agalactiae is a major bacterial pathogen that is the cause of serious economic losses in many species of freshwater, marine and estuarine fish worldwide. A highly efficacious S. agalactiae vaccine was developed using extracellular products (ECP) and formalin-killed whole cells of S.

Evaluation of a recombinant salivary gland protein (thrombostasin) as a vaccine candidate to disrupt blood-feeding by horn flies.

The potential for controlling blood-feeding by the cattle pest, Haematobia irritans irritans (horn fly), was tested by vaccination against thrombostasin (TS), an inhibitor of mammalian thrombin that is released into skin during horn fly blood-feeding. The increase in blood meal size that occurred for flies feeding on sensitized non-vaccinated hosts was blocked and egg development in female flies was delayed when horn flies fed on rabbits and cattle immunized with recombinant TS. This demonstration of the impact of disrupting TS action by vaccination provides a novel approach toward control of this veterinary pest and offers a paradigm for limiting blood-feeding in other medically-important insect species.

Molecular variability of house finch Mycoplasma gallisepticum isolates as revealed by sequencing and restriction fragment length polymorphism analysis of the pvpA gene.

Mycoplasma gallisepticum, a major pathogen of chickens and turkeys, has caused significant declines in house finch (Carpodacus mexicanus) populations in the eastern United States since it was first observed in this species in 1994. There is evidence that M. gallisepticum infection is now endemic among eastern house finches, although disease prevalence has declined, suggesting an evolving host-parasite relationship.

Trinucleotide GAA repeats dictate pMGA gene expression in Mycoplasma gallisepticum by affecting spacing between flanking regions.

The pMGA genes of the avian respiratory pathogen Mycoplasma gallisepticum encode a family of hemagglutinins that are subject to phase variation. A trinucleotide GAA repeat region is located upstream of the pMGA transcription start site. The length of the repeat region varies at a high frequency due to changes in the number of repeat units.

Transposon mutagenesis of Mycoplasma gallisepticum by conjugation with enterococcus faecalis and determination of insertion site by direct genomic sequencing.

Few genetic systems for studying mycoplasmas exist, but transposon Tn916 has been shown to transpose into the genomes of some species and can be used as an insertional mutagen. In the current study, the ability of Enterococcus faecalis to serve as a donor for the conjugative transfer of transposon Tn916 into the genome of the avian pathogen Mycoplasma gallisepticum strain PG31 was examined. Transconjugants were obtained at a frequency of > or =6 x 10(-8) per recipient CFU.

Phenotypic variation of Mycoplasma iowae surface antigen.

The diseases caused by mycoplasmas are generally chronic and persistent and apparently occur notwithstanding a host immune response. The ability to evade the host immune response is facilitated by phenotypic variation in the mycoplasma surface components. The avian mycoplasmas Mycoplasma gallisepticum and Mycoplasma synoviae have both been previously described to be able to switch their surface antigens, and recent evidence indicates that M.

GAA trinucleotide repeat region regulates M9/pMGA gene expression in Mycoplasma gallisepticum.

Mycoplasma gallisepticum, the cause of chronic respiratory infections in the avian host, possesses a family of M9/pMGA genes encoding an adhesin(s) associated with hemagglutination. Nucleotide sequences of M9/pMGA gene family members indicate extensive sequence similarity in the promoter regions of both the transcribed and silent genes. The mechanism that regulates M9/pMGA gene expression is unknown, but studies have revealed an apparent correlation between gene expression and the number of tandem GAA repeat motifs located upstream of the putative promoter.

Adhesion inhibition of Mycoplasma iowae to chicken lymphoma DT40 cells by monoclonal antibodies reacting with a 65-kD polypeptide.

Tissue- and cell-specific attachment of mycoplasmas is a key aspect of the host-parasite relationship. In this study, monoclonal antibodies (MAbs) recognizing surface membrane polypeptides with molecular masses of 46 kD (p46) and 65 kD (p65), respectively, were examined in a microtiter cell attachment (agglutination) inhibition assay. MAbs MI3, MI6, and MI12 reacting with p65 polypeptide of Mycoplasma iowae inhibited attachment of the organisms to chicken lymphoma (DT 40) cells.

A protein (M9) associated with monoclonal antibody-mediated agglutination of Mycoplasma gallisepticum is a member of the pMGA family.

A 62-kDa cell surface antigen (M9) of Mycoplasma gallisepticum PG31 that mediates antibody-induced agglutination of the organism was purified and subjected to N-terminal amino-acid sequencing. A 999-bp region of the cDNA encoding the M9 protein was generated by reverse transcription-PCR, and its nucleotide sequence was determined. PCR primers based on this sequence were used to screen a genomic DNA library of PG31.

Inactivation of rotavirus by new polymeric water disinfectants.

Two new insoluble polymeric materials were evaluated for their efficacies in inactivating rotavirus in flowing water in a biocidal filter application. The two polymers are N-chloro and N-bromo derivatives of a poly-styrene hydantoin prepared from commercial poly-styrene. The studies were conducted for rotavirus in halogen demand-free water at pH 7.

A platelet activation-specific monoclonal antibody that recognizes a receptor-induced binding site on canine fibrinogen.

An activation-specific monoclonal antibody (MoAb) termed "Canine Activated Platelet 1" (CAP1) has been developed and partially characterized. Flow cytometric studies of isolated canine platelets, using adenosine diphosphate (ADP) and platelet activating factor (PAF) as agonists, demonstrated that CAPI binding site number was proportional to agonist strength and agonist concentration. MoAb CAP1 binding was diminished by ethylenediamine-tetraacetic acid, suggesting that the antigen was either stabilized by calcium or antigen binding to the platelet surface was mediated by calcium.

Production and characterization of monoclonal antibodies against avian reovirus strain S1133.

Monoclonal antibodies (MAbs) were produced against the avian reovirus strain S1133. MAbs were isotyped and used to develop diagnostic tests. Splenocytes from immunized mice were screened by enzyme-linked immunosorbent assay (ELISA).

Chronic inflammatory demyelinating polyneuropathy in dogs and cats.

Over the past several years, we have accumulated data on a spontaneous demyelinating peripheral neuropathy that is not well identified in domestic animals. This disorder occurs in dogs and cats of either sex and does not appear breed-related. Onset of signs is usually insidious and the course is typically chronic, sometimes relapsing, and often slowly progressive.

Identification and characterization of a Mycoplasma synoviae 55,000-molecular-weight antigen associated with hemagglutination.

Serological studies have shown that some antigenic determinants are conserved among several pathogenic Mycoplasma species, including Mycoplasma pneumoniae, M. genitalium, and M. gallisepticum.

Cloning and partial sequence analysis of a Mycoplasma synoviae DNA fragment encoding epitopes shared with the major adhesin P1 protein of Mycoplasma pneumoniae.

Polyclonal antibodies specific for the adhesin P1 protein of Mycoplasma pneumoniae were used to screen an expression library of M. synoviae genomic DNA constructed in the expression vector lambda gt11. Following several cycles of immunoscreening using the anti-P1 antiserum, a lambda gt11 recombinant clone containing 3.

Mycoplasma corogypsi sp. nov., a new species from the footpad abscess of a black vulture, Coragyps atratus.

Strain BV1 was isolated from the exudate of the footpad abscess of a black vulture (Coragyps atratus). The colonies had a "fried-egg" appearance consistent with that of mycoplasmal species. Electron microscopic examination of the cells revealed irregular elongated or elliptical forms and smaller circular budding processes.

Production and characterization of monoclonal antibodies against variant A infectious bursal disease virus.

Monoclonal antibodies (MAbs) were produced against a variant subtype of infectious bursal disease virus (IBDV) from Delaware variant isolate A (Var-A). Splenocytes from immunized mice were fused to myeloma cells, and antibody-producing hybridomas were screened by dot-blot enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence (IF) against the homologous isolate. Specificity of the MAbs was tested against viral isolates representing all six serologic subtypes of IBDV and three untyped IBDVs--GLS, Ark, and Miss--found in serotype 1 by dot-blot ELISA and IF.

Antigenic variation of Mycoplasma gallisepticum, as detected by use of monoclonal antibodies.

A panel of monoclonal antibodies (MAb) developed against Mycoplasma gallisepticum strain PG31 was used to probe the antigenic profiles of 5 recognized strains (PG31, R, S6, F, A5969) and 6 field isolates of M gallisepticum. Monoclonal antibody G9 predominantly recognized antigens at apparent molecular mass positions of 90 to 98 kDA. The MAb reacted with all strains and isolates, but the molecular mass position of the antigens varied among some mycoplasmas.