ABSTRACT
PURPOSE: Ocular hypotony after trabeculectomy may be treated medically, surgically and with a tamponade. Three cases are reported in which a scleral lens was applied to treat ocular hypotony after mitomycin C (MMC) augmented trabeculectomy. METHODS: In this retrospective case series the records of three eyes of three patients who developed ocular hypotony after they had undergone trabeculectomy augmented with MMC were evaluated. The patients were between 11 and 69 years of age and the intraocular pressure (IOP) after surgery ranged between 3 and 6 mmHg. All three patients showed a negative Seidel test; one had suspected hypotonic maculopathy and one had a collapsed anterior chamber. After unsuccessful treatment with large bandage lenses all three patients were subsequently fitted with a scleral lens. The scleral lens was fitted to fully cover and compress the bleb. Scleral lenses were worn continuously with a check-up after one night of wear and subsequent check-ups when needed. One patient continued to wear the scleral lens for a further 6.5 months on a daily wear basis. RESULTS: In all three eyes the IOP was higher after wearing the scleral lens. Two patients stopped wearing the scleral lens after the IOP was stable. One patient developed a cataract; the cataract surgery was combined with a bleb revision and scleral lens wear was therefore discontinued. DISCUSSION: The scleral lens might be a useful tool in the treatment of ocular hypotony after trabeculectomy augmented MMC surgery. The effect of the scleral lens on the ocular pressure is unpredictable. Caution is advised in vulnerable corneas due to risk factors such as hypoxia and infection. Further research is warranted to establish the safety of the procedure, the patient selection and the overall success in a larger patient group.
Subject(s)
Contact Lenses , Ocular Hypotension/therapy , Sclera , Trabeculectomy/adverse effects , Adolescent , Aged , Alkylating Agents/administration & dosage , Child , Female , Humans , Intraocular Pressure/physiology , Male , Mitomycin/administration & dosage , Ocular Hypotension/etiology , Prosthesis Fitting , Retrospective Studies , Tonometry, Ocular , Visual AcuityABSTRACT
Cowdria ruminantium is a rickettsial parasite which causes heartwater, a economically important disease of domestic and wild ruminants in tropical and subtropical Africa and parts of the Caribbean. Because existing diagnostic methods are unreliable, we investigated the small-subunit ribosomal RNA (srRNA) gene from heartwater-infected material to characterise the organisms present and to develop specific oligonucleotide probes for polymerase chain reaction (PCR) based diagnosis. DNA was obtained from ticks and ruminants from heartwater-free and heartwater-endemic areas from Cowdria in tissue culture. PCR was carried out using primers designed to amplify only rickettsial srRNA genes, the target region being the highly variable V1 loop. Amplicons were cloned and sequenced; 51% were C. ruminantium sequences corresponding to four genotypes, two of which were identical to previously reported C. ruminantium sequences while the other two were new. The four different Cowdria genotypes can be correlated with different phenotypes. Tissue-culture samples yielded only Cowdria genotype sequences, but an extraordinary heterogeneity of 16S sequences was obtained from field samples. In addition to Cowdria genotypes we found sequences from previously unknown Ehrlichia spp., sequences showing homology to other Rickettsiales and a variety of Pseudomonadaceae. One Ehrlichia sequence was phylogenetically closely related to Ehrlichia platys (Group II Ehrlichia) and one to Ehrlichia canis (Group III Ehrlichia). This latter sequence was from an isolate (Germishuys) made from a naturally infected sheep which, from brain smear examination and pathology, appeared to be suffering from heartwater; nevertheless no Cowdria genotype sequences were found in this isolate. In addition no Cowdria sequences were obtained from uninfected ticks. Complete 16S rRNA gene sequences were determined for two C. ruminantium genotypes and for two previously uncharacterised heartwater-associated Ehrlichia spp. Sequenced difference within the V1 loop were sufficient for the derivation of four Cowdria genotype-specific oligonucleotide probes. Four further probes were designed; one for the detection of any Cowdria genotype, one for the detection of any Group II Ehrlichia sp., one for any Group III Ehrlichia sp. and one for all Pseudomonadaceae. All the probes were specific except that for the Cowdria (Ball 3) genotype. The high prevalence (96%) in field samples of pseudomonad-like 16S sequences was the result of environmental contamination. The probes were used to screen DNA from goats in an area free of both Amblyomma ticks and clinical heartwater. A substantial proportion (42%) gave positive reactions for the apparently apathogenic Cowdria (Omatjenne), indicating that this genotype is relatively common.
Subject(s)
Ehrlichia ruminantium/genetics , Ehrlichia ruminantium/isolation & purification , Heartwater Disease/microbiology , RNA, Ribosomal/genetics , Animals , Base Sequence , DNA Primers , Ehrlichia/genetics , Genes, Bacterial , Genotype , Goats , Mammals , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , Probability , Restriction MappingABSTRACT
Although the silicone rubber contact lens (SRCL) is not used frequently, there area number of clinical indications for its use which include paediatric and adult aphakia, decompensated cornea, dry eye, irregular cornea, eyelid defects, corneal ulcer and corneal perforation. The properties of silicone rubber are reviewed and the fitting technique of the SRCL is described.
Subject(s)
Ehrlichia ruminantium/classification , Ehrlichia/genetics , Heartwater Disease/microbiology , Alphaproteobacteria/classification , Alphaproteobacteria/genetics , Animals , Base Sequence , DNA Primers , Ehrlichia/classification , Ehrlichia/pathogenicity , Ehrlichia ruminantium/genetics , Ehrlichiosis/microbiology , Ehrlichiosis/veterinary , Heartwater Disease/diagnosis , Humans , Molecular Sequence Data , Phylogeny , Polymerase Chain Reaction , RuminantsABSTRACT
The competitive inhibition ELISA (CI-ELISA) format overcomes problems associated with antigen purity since the specificity of the CI-ELISA depends solely on the monoclonal antibody (mAb) used. Therefore, the CI-ELISA format is well suited for use with recombinant antigens. Molecular clones expressing a conserved 19 kDa protein of Anaplasma marginale and a 34 kDa protein of Babesia equi were derived and characterized. The 19 kDa A. marginale protein, conserved in all recognized Anaplasma species, and present in the infected tick salivary gland, was reactive with all bovine immune sera tested. The 34 kDa B. equi protein contains a protein epitope bound by antibody in equine immune sera from 19 countries. Monoclonal antibodies reactive with these proteins were derived and applied with recombinant copies of the 19 kDa A. marginale and 34 kDa B. equi proteins in a CI-ELISA format.
Subject(s)
Anaplasmosis/diagnosis , Antigens, Bacterial/immunology , Babesiosis/diagnosis , Enzyme-Linked Immunosorbent Assay/methods , Amino Acid Sequence , Animals , Cattle , Conserved Sequence , Salivary Glands/microbiology , Serologic Tests , Ticks/microbiologyABSTRACT
Immunization with Anaplasma marginale outer membranes induced immunity against clinical disease which correlated with antibody titer to outer membrane proteins, including a 19-kDa protein (N. Tebele, T. C. McGuire, and G. H. Palmer, Infect. Immun. 59:3199-3204, 1991). This 19-kDa protein, designated major surface protein 5 (MSP-5), was encoded by a single-copy 633-bp gene. The molecular mass of MSP-5, defined in immunoblots by binding to monoclonal antibody ANAF16C1, was conserved among all recognized species of Anaplasma: A. marginale, A. centrale, and A. ovis. Recombinant MSP-5, which absorbed the antibody reactivity of bovine immune serum to native MSP-5, was recognized by anti-A. marginale and anti-A. centrale immune sera in a competitive inhibition assay with monoclonal antibody ANAF16C1. The presence of antibody to the epitope defined by monoclonal antibody ANAF16C1 in all postinfection sera tested indicates that this epitope is a potential diagnostic antigen for use in identifying persistently infected cattle.
Subject(s)
Anaplasma/genetics , Bacterial Outer Membrane Proteins/genetics , Genes, Bacterial , Amino Acid Sequence , Animals , Antibodies, Monoclonal/immunology , Bacterial Outer Membrane Proteins/immunology , Base Sequence , Cattle , Cloning, Molecular , Epitopes , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Molecular WeightABSTRACT
An Anaplasma centrale genomic library was constructed in pUC13. Two clones pAC5 and pAC137 hybridising to A. centrale and A. marginale DNA were isolated from this library. One of these, pAC5, also hybridised to DNA from A. ovis. The total insert of pAC5 was subcloned into pBR322. This subclone, pAC5-12, could detect 1 ng A. centrale, 0.5 ng A. marginale and 3.9 ng A. ovis DNA. The hybridisation pattern obtained with pAC5-12 on digests of A. centrale, A. marginale and A. ovis DNA suggests that this probe detects EcoR1 and Hind111-polymorphisms. Probe pAC5-12 could detect A. ovis DNA in 36% of blood samples tested compared to the 33% detectability obtained with microscopy.
Subject(s)
Anaplasma/genetics , Anaplasmosis/diagnosis , DNA Probes , DNA, Bacterial/blood , Anaplasma/isolation & purification , Animals , Blotting, Southern , Cattle , Gene Library , Nucleic Acid Hybridization , Restriction MappingABSTRACT
DNA from the Washington, South-Idaho, Virginia and Florida isolates of Anaplasma marginale was hybridized to probes specific for Anaplasma centrale and A. marginale. The A. centrale probes AC-2 and AC-4 hybridized to identical bands on all of these isolates. The hybridization patterns suggests that the Virginia, Florida and the South African isolates are similar. A number of bands were obtained with the Washington isolate which differed from those obtained with the other isolates. Probe AC-2 could be developed to identify relatedness among Anaplasma isolates. Probe AC-2 detected A. marginale DNA in midgut material from infected Dermacentor andersoni ticks. No hybridization was obtained with DNA from salivary gland tissues from these infected ticks.
Subject(s)
Anaplasma/isolation & purification , DNA Probes , DNA, Bacterial , Dermacentor/microbiology , Nucleic Acid Hybridization , Ticks/microbiology , Anaplasma/genetics , AnimalsABSTRACT
Anaplasmosis can be diagnosed either by immunological techniques or by direct microscopic examination of blood smears. Both methods are time-consuming and labour intensive. The use of DNA probes in an hybridization assay may simplify the diagnosis of anaplasmosis in cattle and sheep. A genomic DNA library of Anaplasma centrale was constructed in an expression vector and screened to detect clones containing A. centrale DNA. Four probes which hybridized to A. centrale and Anaplasma marginale DNA were isolated. One of these (AC-1) hybridized only to A. centrale DNA, whereas AC-2, AC-3 and AC-4 could detect DNA from both A. centrale and A. marginale. Probes AC-1 and AC-2 could detect 127 ng and 8 ng DNA respectively, while AC-3 and AC-4 detected 64 ng A. centrale DNA.
