Layout proposal for a microtitre plate to be used in routine antimicrobial susceptibility testing of bacteria from infections of dogs and cats

The determination of minimum inhibitory concentrations by broth microdilution is recommended as method of choice for susceptibility testing of veterinary bacterial pathogens. Accordingly, broth microdilution is used in veterinary routine diagnostic laboratories at a progressive rate. To reduce the costs of susceptibility testing, it is reasonable to develop widely accepted uniform microtitre plate layouts that are produced in large quantities. Such microtitre plate layouts have already been developed and published for the susceptibility testing of pathogens from food-producing animals. However, a microtitre plate layout, especially designed for the testing of bacteria from dogs and cats, should be available, too. The choice of the antimicrobial agents or combinations of antimicrobial agents to be included in a suitable layout should be based on the following criteria: (1) the approval and availability of an antimicrobial agent or combination of agents, (2) known cross-resistances, and (3) availability of approved clinical breakpoints. The latter point is of particular importance for the choice of the numbers of concentrations per antimicrobial agent tested and the range of test concentrations. Taking into account these aspects, a science-based layout proposal for microtitre plates, which are suitable for routine testing of bacteria from dogs and cats, is presented and discussed.     

Pharmacokinetics of pradofloxacin and doxycycline in serum, saliva, and tear fluid of cats after oral administration

The pharmacokinetic properties of pradofloxacin and doxycycline were investigated in serum, saliva, and tear fluid of cats. In a crossover study design, six cats were treated orally with a single dose of pradofloxacin (Veraflox^® Oral Suspension 2.5%) and doxycycline (Ronaxan^® 100 mg) at 5 mg/kg body weight. Following administration, samples of serum, saliva, and tear fluid were taken in regular intervals over a period of 24 h and analysed by turbulent flow chromatography/tandem mass spectrometry. All values are given as mean ± SD. Pradofloxacin reached a mean maximum serum concentration ( C _max) of 1.1 ± 0.5 μg/mL after 1.8 ± 1.3 h ( t _max). In saliva and tear fluid, mean C _max was 6.3 ± 7.0 and 13.4 ± 20.9 μg/mL, respectively, and mean t _max was 0.5 ± 0 and 0.8 ± 0.3 h, respectively. Doxycycline reached a mean C _max in serum of 4.0 ± 0.8 μg/mL after 4.3 ± 3.2 h. Whilst only at two time-points doxycycline concentrations close to the limit of quantification were determined in tear fluid, no detectable levels were found in saliva. The high concentrations of pradofloxacin in saliva and tear fluid are promising to apply pradofloxacin for the treatment of conjunctivitis and upper respiratory tract infections in cats. As doxycycline is barely secreted into these fluids after oral application the mechanisms of its clinical efficacy remain unclear.     

Investigation of Chlamydophila spp. in dairy cows with reproductive disorders

Background

Reports worldwide indicate high prevalence of Chlamydophila spp. infection in cattle. To assess the prevalence in Sweden, 525 cows in 70 dairy herds with reproductive disorders was investigated.

Methods

To detect antibodies two commercially available kits were used. Moreover, 107 specimens, including vaginal swabs, organ tissues and milk were analysed by Polymerase Chain Reaction (PCR).

Results

Two (0.4%) cows were seropositive in the Pourquier Cp. abortus ELISA. The seroprevalence with the Chekit ELISA was 28% with no difference between cases and controls. Five specimens were positive in real-time PCR and further analysed by nested PCR. Cp. pecorum was confirmed by partial omp1 DNA sequencing of the nested PCR product of vaginal swabs from control cows.

Conclusion

The results suggest that Cp. abortus infection is absent or rare in Swedish cows whereas Cp. pecorum is probably more spread. They also suggest that Chlamydophila spp. are not related to reproduction disorders in Swedish cattle.     

Treatment of cats with feline infectious peritonitis

Feline infectious peritonitis (FIP) infection resulting in clinical signs is invariably fatal despite clinical intervention. As FIP is an immune-mediated disease, treatment is mainly aimed at controlling the immune response triggered by the infection with the feline coronavirus (FCoV). Immune suppressive drugs such as prednisone or cyclophosphamide may slow disease progression but do not produce a cure. In nearly every published case report of attempted therapy for clinical FIP, glucocorticoids have been used; there are, however, no controlled studies that evaluate the effect of glucocorticoids as a therapy for FIP. Some veterinarians prescribe immune modulators to treat cats with FIP with no documented controlled evidence of efficacy. It has been suggested that these agents may benefit infected animals by restoring compromised immune function, thereby allowing the patient to control viral burden and recover from clinical signs. However, a non-specific stimulation of the immune system may be contraindicated as clinical signs develop and progress as a result of an immune-mediated response to the mutated FCoV.     

Spectrum analysis of respiratory sounds in exercising horses with experimentally induced laryngeal hemiplegia or dorsal displacement of the soft palate

OBJECTIVE: To record respiratory sounds in exercising horses and determine whether spectrum analysis could be use to identify sounds specific for laryngeal hemiplegia (LH) and dorsal displacement of the soft palate (DDSP). ANIMALS: 5 Standardbred horses. PROCEDURE: Respiratory sounds were recorded and pharyngeal pressure and stride frequency were measured while horses exercised at speeds corresponding to maximum heart rate, before and after induction of LH and DDSP. RESULTS: When airway function was normal, expiratory sounds predominated and lasted throughout exhalation. After induction of LH, expiratory sounds were unaffected; however, all horses produced inspiratory sounds characterized by 3 frequency bands centered at approximately 0.3, 1.6, and 3.8 kHz. After induction of DDSP, inspiratory sounds were unaffected, but a broad-frequency expiratory sound, characterized by rapid periodicity (rattling) was heard throughout expiration. This sound was not consistently detected in all horses. CONCLUSIONS AND CLINICAL RELEVANCE: The technique used to record respiratory sounds was well tolerated by the horses, easy, and inexpensive. Spectrum analysis of respiratory sounds from exercising horses after experimental induction of LH or DDSP revealed unique sound patterns. If other conditions causing airway obstruction are also associated with unique sound patterns, spectrum analysis of respiratory sounds may prove to be useful in the diagnosis of airway abnormalities in horses.     

Comparison of six in-house tests for the rapid diagnosis of feline immunodeficiency and feline leukaemia virus infections

Six rapid tests for the diagnosis of feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) infections which have recently been introduced in Europe for use in small animal practice were compared. Eight hundred serum samples were tested and those reacting FIV-positive in at least one of the tests were confirmed by Western blot, and those reacting FeLV-positive were confirmed by virus isolation. The specificity and sensitivity of each test and the quality of the results produced were compared.     

Cervical neoplasia originating from the vagus nerve in a dog

An eight-year-old intact male Bernese mountain dog was referred with a history of chronic vomiting, coughing and signs of respiratory distress. Other historical findings included lethargy, weight loss and choking. On presentation, clinical findings were Horner's syndrome, ipsilateral laryngeal hemiplegia, coughing, gagging, respiratory distress and vomiting. Lateral cervical radiographs showed ill-defined mineralisation in the soft tissue ventral to the third cervical vertebra, while ultrasonography of the neck revealed a well marginated heterogeneous mass with focal hyperechogenic lesions and acoustic shadowing. Results of an ultrasound-guided fine needle aspirate suggested neoplasia. At necropsy, a large tumour was detected in the ventral cervical region, originating from the right vagosympathetic trunk. In view of the infiltrating pattern, the cellular pleomorphism and the numerous mitoses on histopathological examination, the tumour was classified as a malignant peripheral nerve sheath tumour.     

Abiotic triggers for sea and river lamprey spawning migration and juvenile outmigration in the River Rhine,Germany

The activity patterns of migrating lampreys (sea lamprey Petromyzon marinus and river lamprey Lampetra fluviatilis) are poorly understood. This study uses extensive screening data from the cooling water intake of a nuclear power plant on the River Rhine, Germany, gathered on 1308 days over 46 consecutive months between 2011 and 2014. More than 34,000 lampreys were retrieved, of which the great majority (99.9%) were outmigrating nonfeeding postmetamorphic individuals. The main factors controlling outmigration were season, water temperature and river discharge. The bulk of downstream migration was observed each year in spring at water temperatures between 9 and 12°C, with earlier migrations during significant discharge peaks between December and February. The migration activity of postmetamorphic individuals declined during summer and autumn, regardless of discharge. Turbidity increased the occurrence of postmetamorphic lampreys and supported diurnal migration, which accounted for nearly 20% of all lamprey records. In winter, outmigrating individuals of both species exhibited significantly longer body lengths than those detected during other seasons. Haematophagus postmetamorphic sea lampreys were also retrieved (n = 28) and represent the most upstream records of parasitically feeding sea lamprey detected thus far. Adult lampreys migrating upstream were found mainly in spring, but in very low numbers, indicating high prespawning mortalities. The study points to a challenging future for the protection of lampreys in this area: the long duration and the round‐the‐clock nature of the observed migration demand flexible solutions to realise safe operational windows for water intake stations and to minimise impacts on juvenile lampreys.