Suitability of SSCP-PCR analysis for molecular detection of quinolone resistance in Campylobacter jejuni

Foodborne infections with Campylobacter spp. are increasing, especially antibiotic resistant strains are emerging. Quinolone resistant isolates can cause failure of therapy in severe clinical infections. Molecular characterisation is needed for the detection of resistant variants of C. jejuni. Therefore 23 isolates from poultry and human medicine as well as three control strains were tested for their minimal inhibitory concentration, their Single-Strand-Conformation-Polymorphism (SSCP)-PCR pattern (a method for the detection of resistance determining point mutations), and their sequence of the quinolone resistance determining region (QRDR). Six different SSCP types could be identified: two types for quinolone resistant isolates and other types containing so called silent mutations without influence on the resistance. A genotypic monitoring of the quinolone resistance in C. jejuni can be useful for the early detection of new resistance variants. As a screening method for detection of point mutations in the QRDR the SSCP-PCR can be applied. Compared to other genotypic methods the SSCP-PCR is less time and cost consuming and needs only standard technical equipment.     

Pharmacokinetics and Effects of Alkalization During Oral and Intravenous Administration of Naproxen in Horses

The use of suitable therapeutic protocols is particularly important when extra-label drugs are used or when physiological parameters are modified, as in the case of the administration of alkaline substances to racehorses. The pharmacokinetics of naproxen (NAP), after both intravenous (iv) and oral administration of 10 mg/kg body weight (BW), was investigated in horses under normal metabolic conditions and in horses whose conditions were modified by the iv administration of 250 mg/kg BW of sodium bicarbonate (NaHCO₃). The hypothesis that blood and consequent urinary alkalization could modify NAP pharmacokinetics was evaluated. Drug quantification was performed on serum and urine using High Performance Liquid Chromatography (HPLC) with ultraviolet-visible detection. Results were also integrated with cycloxygenase (COX)-inhibition published data to suggest an appropriate schedule for NAP dosage in horses. After iv administration, NAP was rapidly distributed (t_1/2α: 0.71 ± 0.43 iv NaHCO₃ and 0.55 ± 0.62 hours No NaHCO₃), whereas its elimination was quite slow (t_1/2β: 6.74 ± 0.41 hours), particularly in iv NaHCO₃ animals (t_1/2β: 8.95 ± 1.37 hours). After oral treatments, NAP was more rapidly absorbed and elimination was slower in iv NaHCO₃ animals (t_1/2λ_z: 17.50 ± 6.66 vs. 7.17 ± 0.91 hours). The oral bioavailability of NAP was approximately 87% and 77% in No NaHCO₃ and iv NaHCO₃, respectively. Urinary excretion of the drug as a parent compound was low. The alkalization procedure did not anticipate the elimination of the acidic drug as expected, but it also influenced the absorption of the drug that was administered orally. The dosage scheme of 10 mg/kg BW iv or orally seems to be appropriate to produce an anti-inflammatory effect for 12 to 24 hours.     

Clinical and immunological heterogeneity of canine subepidermal blistering dermatoses with anti‐laminin‐332 (laminin‐5) auto‐antibodies

Laminin‐332 (laminin‐5) is a basement membrane heterotrimeric protein composed of alpha‐3, beta‐3 and gamma‐2 laminin chains. Laminin‐332 polypeptides are targeted by auto‐antibodies in human patients with mucous membrane (cicatricial) pemphigoid or, more rarely, subepidermal vesicular diseases that resemble epidermolysis bullosa acquisita (EBA) or bullous pemphigoid (BP). The objectives of this report were to characterize the clinical, histopathological and immunological characteristics of nine dogs with auto‐antibodies targeting laminin‐332. Immunological investigations consisted of direct immunofluorescence (IF), indirect IF with intact and salt‐split canine gingival, and salt‐split normal or laminin‐332‐deficient human skin, immunoblotting with purified human laminin‐332 and immunoblotting with recombinant NC1 domain of human collagen VII. All dogs exhibited varying degrees of skin blistering and ulceration associated with microscopic subepidermal vesiculation with or without inflammatory cells. Indirect IF established that circulating IgG auto‐antibodies bound the dermal side of salt‐split canine lip and human skin. In five dogs, IgG variably recognized the basement membrane of laminin‐332‐deficient human skin (three dogs negative, two dogs positive). In all nine dogs, IgG auto‐antibodies detected purified human laminin‐332 by immunoblotting. In two dogs, additional targeting of collagen VII‐NC1 was present. These observations establish laminin‐332 as a novel basement membrane antigen in dogs with autoimmune blistering diseases with variable clinical phenotypes. The names ‘acquired junctional epidermolysis bullosa’, ‘anti‐laminin‐332 mucous membrane pemphigoid (MMP)’ and ‘mixed auto‐immune subepidermal blistering dermatosis’ are proposed for dogs with clinical signs reminiscent of EBA, MMP or BP respectively.     

Macrophakia in three cats

This report describes three cases of bilateral macrophakia in 3-, 5-, and 9-year-old cats, respectively. All cats were presented because of visual deficits. Ophthalmic examination revealed macrophakia (the vitreous was replaced by the lens) and retinal changes (tapetal hyper-reflectivity, attenuation of retinal vessels, and retinal folds) in all cats. In addition, bilateral subconjunctival orbital fat prolapse in one cat and microphthalmos in another cat were present. To confirm the ophthalmologic diagnosis of macrophakia, gross pathology examination in one cat and ultrasound examination in another cat were performed.