Biofilm production by pathogens associated with canine otitis externa,and the antibiofilm activity of ionophores and antimicrobial adjuvants

Otitis externa (OE) is a frequently reported disorder in dogs associated with secondary infections by Staphylococcus, Pseudomonas and yeast pathogens. The presence of biofilms may play an important role in the resistance of otic pathogens to antimicrobial agents. Biofilm production of twenty Staphylococcus pseudintermedius and twenty Pseudomonas aeruginosa canine otic isolates was determined quantitatively using a microtiter plate assay, and each isolate was classified as a strong, moderate, weak or nonbiofilm producer. Minimum biofilm eradication concentration (MBEC) of two ionophores (narasin and monensin) and three adjuvants (N‐acetylcysteine (NAC), Tris‐EDTA and disodium EDTA) were investigated spectrophotometrically (OD_570nm) and quantitatively (CFU/ml) against selected Staphylococcus and Pseudomonas biofilm cultures. Concurrently, minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of planktonic cultures were assessed. 16/20 of the S. pseudintermedius clinical isolates were weak biofilm producers. 19/20 P. aeruginosa clinical isolates produced biofilms and were distributed almost equally as weak, moderate and strong biofilm producers. While significant antibiofilm activity was observed, no MBEC was achieved with narasin or monensin. The MBEC for NAC ranged from 5,000–10,000 µg/ml and from 20,000–80,000 µg/ml against S. pseudintermedius and P. aeruginosa, respectively. Tris‐EDTA eradicated P. aeruginosa biofilms at concentrations ranging from 6,000/1,900 to 12,000/3,800 µg/ml. The MBEC was up to 16‐fold and eightfold higher than the MIC/MBC of NAC and Tris‐EDTA, respectively. Disodium EDTA reduced biofilm growth of both strains at concentrations of 470 µg/ml and higher. It can be concluded that biofilm production is common in pathogens associated with canine OE. NAC and Tris‐EDTA are effective antibiofilm agents in vitro that could be considered for the treatment of biofilm‐associated OE in dogs.     

Thymic hyperplasia causing right ventricular outflow tract compression following treatment for oesophageal rupture in an Arabian colt

True thymic hyperplasia has been reported sporadically in the human literature as an immunological rebound phenomenon following immunosuppressive treatment or disease. There are limited accounts in domestic species, mainly following vaccination, and thymic hyperplasia has not been a recognised condition in the horse to date. This report details a case of true thymic hyperplasia in a 10-week-old Arabian colt diagnosed by histopathology of core biopsy samples. The colt developed pulmonary stenosis caused by compression due to a space occupying lesion in the cranial mediastinum following a 3-month history of hospitalisation for treatment of traumatic oesophageal rupture with perioesophageal abscess formation. Diagnostic imaging of the cranial mediastinum was indicative of a thymic mass, and histopathology confirmed the mass was normal (hyperplastic) thymic tissue. The colt was treated with a tapering dose of corticosteroids, which led to involution of the hyperplastic tissue and resolution of pulmonary artery compression. Thymic hyperplasia may be an unrecognised sequela to chronic inflammation in horses and was only identified in this case when the size was sufficient to compress right cardiac outflow.     

The Neospora caninum-Spain 7 isolate induces placental damage, fetal death and abortion in cattle when inoculated in early gestation

The Nc-Spain 7 isolate of Neospora caninum, which was newly obtained from an asymptomatic congenitally infected calf, demonstrated a similar virulence as Nc-1 strain in mouse models. The aim of this study was to characterize the pathogenesis of Nc-Spain 7 isolate in cattle after experimental infection at 65 days of gestation. For this purpose, thirteen pregnant heifers were divided into three groups as follows: group A: 7 heifers inoculated with 1×10(8) tachyzoites of Nc-Spain 7 isolate; group B: 4 heifers inoculated with 1×10(8) tachyzoites of Nc 1 strain; and group C: 2 heifers received PBS. Serum samples were collected weekly and heparinized blood samples were collected three times (0, 28 and 42 days after inoculation) by jugular venipuncture. Placenta and fetal tissue samples were collected at time of necropsy. Specific antibody response in the dams was tested by IFAT, indirect ELISA, and rNcGRA7 and rNcSAG4 based-ELISA. Specific antibody response in fetal fluids was tested by IFAT. IFN-γ production was measured after in vitro culture of PBMC and the supernatant was assessed using a commercial kit (BOVIGAM). A significant increase in N. caninum antibody responses was detected in groups A and B by IFAT and by i-ELISA from day 14 after inoculation onwards. Besides, antibody response against rNCGra7 protein was also detected in all inoculated heifers by rNcGra7-based ELISA. Four fetuses from group A and one from group B were aborted between 3 and 5 weeks after infection. In the recovered fetuses, only 3 out of 4 fetal fluids from fetuses of group A and 1 out of 3 of group B were seropositive by IFAT, but all of them were positive by PCR. Transplacental transmission could be determined in all fetuses from groups A and B by PCR and/or IHC. Heifers of group C and their fetuses remained negative by all techniques. The results of this study demonstrate that the NC-Spain 7 isolate could be transmitted transplacentally, and produced fetal death and abortion in cattle.     

Temperament and hypothalamic-pituitary-adrenal axis function are related and combine to affect growth, efficiency, carcass, and meat quality traits in Brahman steers

Associations between temperament, stress physiology, and productivity were studied in yearling Brahman steers (n = 81). Steers differed in calpain system gene marker status; 41 were implanted with a hormonal growth promotant at feedlot entry. Temperament was assessed with repeated measurements of flight speed (FS) and crush score (CS) during 6 mo of backgrounding at pasture and 117 d of grain finishing. Adrenal responsiveness was assessed with ACTH challenge, with plasma samples collected immediately before and 60 min after challenge. Steers with higher FS and CS had higher prechallenge plasma cortisol, glucose, lactate, and nonesterified fatty acid concentrations. The ACTH-induced cortisol response was unrelated to FS or CS, but glucose remained higher after challenge in flightier steers. The hormonal growth promotant reduced adrenal responsiveness; tenderness genotype had no effect. When temperament assessments and cortisol concentrations before and after challenge were combined in a principal components analysis, four vectors accounting for 38%, 25%, 18%, and 9% of the variation were identified. The first vector had significant loadings on temperament and prechallenge cortisol; increasing scores were associated with increased plasma glucose, lactate, and nonesterified fatty acid and with reductions in BW and feedlot growth rates, carcass fatness, and muscle pH. The second vector loaded only on ACTH-induced cortisol response; increased scores related to increased residual feed intake, number of daily feed sessions, and meat marbling score. The third and fourth vectors had different loadings on FS and CS and appeared to identify different aspects of temperament measured by FS or CS. Fewer associations were found between the third or fourth vectors and productivity traits, possibly because of lower variance accounted for by these vectors. In conclusion, temperament was related to prechallenge cortisol but not to ACTH-induced cortisol response. Principal components analysis separated these traits into separate components, which in turn had different relations with productivity traits. The largest component of temperament was described similarly by FS and CS, but there were smaller components that these described differently. There were some temperament-related differences in the metabolic status of the steers which were not related to the variation in cortisol, suggesting involvement of the sympatho-adrenal-medullary axis in these temperament-related effects.