Isolation and identification of Treponema pedis and Treponema phagedenis-like organisms from bovine digital dermatitis lesions found in dairy cattle in Turkey.

The isolation and identification of microorganisms associated with digital dermatitis (DD) in Turkish dairy cattle was investigated using punch skin biopsy samples from typical clinical lesions; they were collected from dairy farms and abattoirs in 5 different geographical locations in Turkey. Morphological characteristics and flagellation types were examined using a transmission electron microscope, and their enzyme profiles by enzyme activation kits; their catalase reaction characteristics were evaluated by the addition of 3% H2O2. Their phylogenies were identified using 16S rRNA and the results compared with known gene bank data. Bacterial cells were 5.0 to 18.2 µm long, 0.2 to 0.5 µm wide, and their minimum number of periplasmic flagellum was 4 (4:8:4) with a maximum of 8 (8:16:8). All isolates were catalase negative. Of the spirochetes isolated and identified, group I organisms showed close similarity with Treponema pedis (99% genetic homology), whereas those in group II were similar to Treponema phagedenis (98% homology). This is the first report of specific sub-groups of Treponema spp., isolated from Turkish dairy cattle presenting with DD lesions, being associated with this disease; these morphotypes were similar to those found globally in housed dairy cattle units and are probably significant microorganisms associated with the aetiopathogensis of this infectious disease causing acute bovine lameness. These results suggest that the distribution of DD-associated treponemes is not specific to particular geographic regions of Turkey.

Evaluation of preemptive dexketoprofen trometamol effect on blood chemistry, vital signs and postoperative pain in dogs undergoing ovariohysterectomy.

OBJECTIVE: To investigate the postoperative analgesic effects of preemptive dexketoprofen trometamol in dogs subjected to ovariohysterectomy (OHE). MATERIAL AND METHODS: Seventeen adult bitches of various breeds were used in this study. The dogs were randomly allocated into of two groups. Subjects in the dexketoprofen trometamol (DEX) group (n=10), received intravenous (i.v.) dexketoprofen trometamol, 1 mg/kg, 15 minutes before premedication, while those assigned to the control (C) group (n=7) were given no analgesics prior to premedication. Pain level was assessed by two researchers before the administration of anaesthesia (15 minutes before start) and 0, 1, 2, 4 and 6 hours after surgery. A modified University of Melbourne Pain Scale (UMPS) was used to evaluate pain in both groups. RESULTS: Serum cortisol level changed from 0 to 1 h and from 0 to 1 to 4 h were compared between the groups; the increase in the C group was statistically significant. The modified UMPS was applied to both groups at baseline and postoperative 1, 2, 4 and 6 h. According to this test, the values for DEX were significantly lower than controls at 4 and 6 h (p<0.001). CONCLUSION: Stable vital signs with unchanged biochemical parameters on dexketoprofen administration are a promising finding. The clinical advantage shown by the pain scale difference and the low serum cortisol levels should qualify dexketoprofen for preemptive pain management in dogs (Tab. 5, Fig. 2, Ref. 30).

Comparison of the effects of lidocaine and fentanyl in epidural anesthesia in dogs.

The study included 12 clinically healthy, adult male dogs of various breeds, admitted to our clinic for castration. After general anesthesia with sevoflurane, we administered epidural fentanyl (1 mcg/kg) to fentanyl group, while lidocaine group was given Lidocaine (3 mg/kg) through epidural administration. When hemodynamic parameters were stabilized, first measurements were recorded at minutes 0, 15, 30, 60 in both groups, which included Heart Rate (HR), body temperature, systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), sodium (Na+), potassium (K+), glucose (GLC), and hemoglobin (HB) measurements. In addition, serum samples were obtained from arterial blood at the same measurement times, and pH, pO2, pCO2, HCO3, %O2 Saturation, BE levels were measured. For hematological analysis, WBC, RBC, HCT, THR counts were performed. For serum biochemical analysis, venous blood samples were collected at minutes 0 and 60 and CK, TP, UREA, ALT, AST, ALB, GGT, CRE, CK-MB parameters were assessed using auto-analyzer. Moreover, cortisol levels were measured in the samples collected at minutes 0, 30, and 60.Mean arterial blood pressure values measured at minutes 15, 30 and 60 were found significantly lower in the fentanyl group (p<0.01). In conclusion, we suggest that epidural anesthesia with lidocaine and fentanyl can provide an effective and safe option in high-risk groups (Tab. 5, Fig. 1, Ref. 24).

Effects of abdominal compartment syndrome on gastric emptying time.

INTRODUCTION: Increase in intra-abdominal pressure may affect gastric emptying time but the precise effect has not been demonstrated. Effects of acute increase of intra-abdominal pressure on gastric emptying time can not be demonstrated in clinical or experimental studies. In this study we aimed to study the effect of increased intra-abdominal pressure on gastric emptying time. METHODS: Six male stray dogs that weighed 20-25 kg were studied. Following the induction of general anaesthesia, an abdominal catheter was placed and intra-abdominal pressure was raised at a rate of 5 cm H2O every 10 minutes using intra-abdominal administration of physiologic saline solution until 45 cm H2O pressure was reached. Gastric emptying time was measured scintigraphically at the beginning of the study (IAB 0 cm H2O ) and again four hours later when pressure reached the maximum value (IAP was 45 cm H2O). RESULTS: Gastric emptying time for baseline pressure was in average 51.83±13.16 whereas for 45 cm H2O pressure it was in average 90.83±26.96. This difference was found statistically significant (p<0.05). The differences between baseline values and values after increased intra-abdominal pressure were statistically significant (Tab. 4, Fig. 1, Ref. 26).

The ovarian and uterine arteries in the chinchilla (Chinchilla lanigera).

The purpose of this study was to describe arteries supplying the ovaries and uterus in the chinchilla. Five healthy adult female chinchillas were used. In order to reveal the arterial network by dissecting under a stereoscopic microscope, latex coloured with red ink was injected through the common carotid artery. The ovaries of the chinchilla are supplied by the arteriae ovaricae which formed end-to-end anastomoses with the cranial termination of the arteria uterina. Soon after leaving the aorta abdominalis, the arteriae ovaricae extended 2-3 mm caudolaterally, then released 1 branch and extended caudally and bifurcated into 2 further branches. One of these supplied branches to fat tissue. The other branch coursed caudally and anastomosed with the arteria circumflexa ilium profunda and dispersed into fat tissue. The arteria ovarica further subdivided into 2 rami ovaricae. The origins of the uterine arteries were exclusively from the left arteria iliaca externa. The arteria uterina gave a branch to the arteria umbilicalis and consecutive branches which supplied to the ureter, urinary bladder and cranial aspects of the vagina. It also gave rise to 2-3 branches to the cervix and further supplied 10-12 meandering branches to the uterine horns. The arteria uterina gave rise to many tortuous arteries to the uterus and provided 2 further branches to the ovary.

Diagnostic assessment without cut-offs: application of serology for the modelling of bovine digital dermatitis infection.

Bovine digital dermatitis (BDD) is an epidermitis which is a leading cause of infectious lameness. The only recognized diagnostic test is foot inspection, which is a labour-intensive procedure. There is no universally recognized, standardized lesion scoring system. As small lesions are easily missed, foot inspection has limited diagnostic sensitivity. Furthermore, interpretation is subjective, and prone to observer bias. Serology is more convenient to carry out and is potentially a more sensitive indicator of infection. By carrying out 20 serological assays using lesion-associated Treponema spp. isolates, three serogroups were identified. The reliability of the tests was established by assessing the level of agreement and the concordance correlation coefficient. Subsequently, an ELISA suitable for routine use was developed. The benchmark of diagnostic test validation is conventionally the determination of the key test parameters, sensitivity and specificity. This requires the imposition of a cut-off point. For serological assays with outcomes on a continuous scale, the degree by which the test result differs from this cut-off is disregarded. Bayesian statistical methodology has been developed which enables the assay result also to be interpreted on a continuous scale, thereby optimizing the information inherent in the test. Using a cross-sectional study dataset carried out on 8 representative dairy farms in the UK, the probability of infection, P(I), of each individual animal was estimated in the absence of a 'Gold Standard' by modelling I as a latent variable which was determined by lesion status, L as well as serology, S. Covariate data (foot hygiene score and age) were utilized to estimate P(L) when no lesion inspection was performed. Informative prior distributions were elicited where possible. The model was utilized for predictive inference, by computing estimates of P(I) and P(L) independently of the data. A more detailed and informative analysis of the farm-level distribution of infection could thus be performed. Also, biases associated with the subjective interpretation of lesion status were minimized. Model outputs showed that young stock were unlikely to be infected, whereas cows tended to have high or low probabilities of being infected. Estimates of probability of infection were considerably higher for animals with lesions than for those without. Associations were identified between both covariates and probability of infection in cows, but not in the young stock. Under the condition that the model assumptions are valid for the larger population, the results of this work can be generalized by predictive inference.

Can dantrolene protect spinal cord against ischemia/reperfusion injury? An experimental study.

BACKGROUND: The aim of this experimental study was to investigate the possible protective effect of dantrolene on neuronal injury induced by aortic ischemia/reperfusion (I/R). METHODS: Nineteen rabbits were divided into three groups: sham (group 1, n = 5, no I/R), control (group 2, n = 7, only I/R) and dantrolene (group 3, n = 7, dantrolene + I/R). Abdominal aortic occlusion between the renal arteries and iliac bifurcations was carried out for 30 min. The spinal cord functions of the subjects were assessed using the Tarlov Scale. Blood and cord tissue samples were taken for biochemical and histopathological evaluation. RESULTS: Tarlov scores in group 3 were significantly higher than in group 2 ( P < 0.05). In group 3, the MDA levels of the spinal cord decreased significantly compared to those of group 2 ( P < 0.05). In rabbits with I/R (group 2), the GSH levels of the spinal cord decreased significantly compared to those of group 1 ( P < 0.01), but dantrolene pretreatment significantly prevented a decrease in GSH levels. Histopathological examination showed that group 3 had less vascular proliferation, hemorrhage, edema and neuron loss than group 2. CONCLUSIONS: It was concluded that dantrolene applied after ischemia might help protect the spinal cord against ischemia/reperfusion injury.

Gross morphological features of plexus brachialis in the chinchilla (Chinchilla lanigera).

This study documents the detailed features of the morphological structure and the innervation areas of the plexus brachialis in the chinchilla (Chinchilla lanigera). The animals (5 female and 5 male) were euthanased with ketamine hydrocloride and xylazine hydrocloride combination, 60 mg/kg and 6 mg/kg, respectively. Skin, muscles and nerves were dissected under a stereo-microscope. The brachial plexus of the chinchilla is formed by rami ventrales of C5-C8, T1 and T2, and possesses a single truncus. The subscapular nerve is formed by the rami of the spinal nerves originating from C6 (one thin ramus) and C7 (one thick and 2 thin rami). These nerves innervate the subscapular and teres minor muscles. The long thoracic nerve, before joining with the brachial plexus, obtains branches from C6 and C7 in 5 cadavers (3 male, 2 female), from C7 in 4 cadavers (2 male, 2 female) and from C6-C8 in only 1 female cadaver. These nerves disperse in variable combinations to form the extrinsic and intrinstic named, nerves of the thoracic limb. An undefined nerve branch originates from the rami ventrales of C7, C8 and T1 spinal nerves enter the coracobrachial muscle.

Characterization of a spirochaete isolated from a case of bovine digital dermatitis.

AIMS: The aim of the study was to characterize a spirochaete isolated from the lesions of a cow with digital dermatitis (DD). METHODS AND RESULTS: The characterization was on the basis of its light and electron microscopic appearance, enzymic profile and DNA sequence analysis of its flagellin and 16S rRNA genes. The spirochaete was 6-8-microm long and 0.2-0.3 microm in diameter, and possessed seven to eight periplasmic flagella, with three to five helical turns. The enzymic profile of the bacterium resembles, but is not identical to that of Treponema brennaborense. Its flagellin gene sequence was identical to that of Treponema phagedenis but distinct from that of an ovine spirochaete. Analysis of a 1477-bp region of the 16S rRNA genes indicated that this is a Treponema species and that it is indistinguishable from some isolates made from cases of bovine DD in the United States. Finally, electron microscopy revealed the presence of myovirus-like bacteriophage particles in all cultures of the treponeme examined. CONCLUSIONS: The spirochaete isolate was identified as a Treponema species closely related to some isolates from the United States (by 16S rDNA) and to T. phagedenis (by flagellin gene sequence) and is associated with bacteriophage particles. SIGNIFICANCE AND IMPACT OF THE STUDY: The fact that the isolates with the same or very similar 16S rDNA sequences have been obtained from cases of bovine DD in cattle in different countries at different times, lends further support to the hypothesis that treponemes play a role in the pathogenesis of this disease.

Bovine digital dermatitis and severe virulent ovine foot rot: a common spirochaetal pathogenesis.

A potential pathological role for spirochaetes in bovine digital dermatitis (bovine DD) and severe virulent ovine foot rot (SVOFR) has been considered and a treponeme isolate obtained from each disease in the UK. In this work, we have investigated the hypothesis that the two diseases may have a shared (common) spirochaetal aetiology. Experiments were designed to identify serological similarities and differences between the two spirochaetes; an enzyme-linked immunosorbent assay (ELISA) was developed to detect anti-treponeme antibodies in the sera of cows and sheep against the two-treponeme isolates. Sera were further tested for antigen reactivity by Western blotting. Cattle and sheep with bovine DD and SVOFR, respectively, had increased seropositivity rates to both treponeme isolates, with different patterns of reactivity between farms. In some cattle herds, significant correlations were shown between antibodies to bovine DD treponemes and SVOFR treponemes (P<0.001). In other herds, there was no apparent cross reaction, suggesting the presence of more than one treponeme in bovine DD on some farms. There was no significant correlation between the two treponeme isolates when ELISA-tested against 58 sheep sera from SVOFR cases (P>0.05); sheep showed strong evidence of reactivity to one or the other treponeme antigens, but never to both. Western blotting against both treponeme antigens showed that they frequently displayed different antigen epitopes, although some minor bands were common to both organisms. The data suggest that there are a number of spirochaetes in UK farms, which could be involved in the pathogenesis of either bovine DD or SVOFR.

Some relationships between spirochaete infections and digital dermatitis in four UK dairy herds.

Digital dermatitis, presenting as a focal necrotic dermatitis on the skin of cows' feet adjacent to the interdigital space, has been associated with dermal infection by spirochaetes. Lesions cause severe, transient lameness. An enzyme-linked immunoassay (ELISA) was used to measure IgG(2) antibody titres against three different spirochaetes: USA bovine Treponema isolates 1-9185MED and 2-1498, and a UK strain of Borrelia burgdorferi B31. Logistic models were used to describe lesion prevalence in terms of the antibody titres, herd and milking status, year, lactation number, and yield. The antibody titres and milking status (P<0.001) dominated the logistic fits, with the other variables adding little. Generalised linear models were used to describe the antibody titres in terms of the presence or absence of lesions, herd, year, lactation number, and yield. The presence or absence of lesions had the most significant effect for all three antibodies (P<0.001). Neither the herd effects nor the effects due to lactation number or yield showed consistent patterns.

Effect of medetomidine-butorphanol-ketamine anaesthesia and atipamezole on heart and respiratory rate and cloacal temperature of domestic pigeons.

The purpose of this study was to evaluate the sedative-anaesthetic effects of a combination of medetomidine (M, 50 microg per pigeon), butorphanol (B, 50 microg per pigeon) and ketamine (K, 25 mg per pigeon) in domestic pigeons. Eight domestic pigeons (four male and four female, 8-15 months old) were used. The combination of Medetomidine and butorphanol injectable solutions were used to produce sedation. Ten minutes after M + B administration, K was injected. The anaesthetic effects of the drugs were reversed by administration of Atipamazole (AT) at 60 min after K administration. All drugs were injected into the pectoral muscles. The sedative-anaesthetic effects of the M + B-K combination and, alterations in respiratory rate (RR), heart rate (HR), electrocardiographic (ECG) findings and cloacal temperature (CT) were investigated before and 10 min after pre-medication with M + B, at 5, 15, 30, 45 and 60 min during the onset of K anaesthesia and at 1, 5, 10, 20, 30 and 60 min following the administration of AT. The HR and RR of pigeons decreased within 10 min following M + B administration and remained lower until 1st and 5 min of AT injection, respectively. In ECG, no significant alterations in P, Q, R and S-values were observed, however, arhythmia was recorded for three pigeons, which returned to normal values following AT administration throughout the measurement. Cloacal temperature decreased gradually during the anaesthesia from 41.0 to 32.7 degrees C. The drug combination used in this study produced a satisfactory general anaesthesia for seven of the eight pigeons. All pigeons were unconscious within 5 min after K administration as indicated by disappearance of the palpebral and corneal reflexes and lack of reaction to the pain stimuli during the study. The effect of AT administration was observed within 10 min as all pigeons responded partly against stimuli and all reflexes. It is concluded that M + B-K anaesthesia in pigeons is a safe and reliable anaesthetic protocol for surgery.

Isolation and characterisation of a novel spirochaete from severe virulent ovine foot rot.

A novel spirochaete was isolated from a case of severe virulent ovine foot rot (SVOFR) by immunomagnetic separation with beads coated with polyclonal anti-treponemal antisera and prolonged anaerobic broth culture. The as yet unnamed treponeme differs considerably from the only other spirochaete isolated from ovine foot rot as regards morphology, enzymic profile and 16S rDNA sequence. On the basis of 16S rDNA, it was most closely related to another unnamed spirochaete isolated from cases of bovine digital dermatitis in the USA, raising the possibility of cross-species transmission. Further information is required to establish this novel ovine spirochaete as the cause of SVOFR.

Serological evidence of spirochaetal infections associated with digital dermatitis in dairy cattle.

A potentially infectious aetiology for digital dermatitis in dairy cattle was investigated and centred on the possible involvement of spirochaetes. An enzyme-linked immunosorbent assay (ELISA) was developed to detect bovine anti-Borrelia burgdorferi (B31) and anti-Treponeme (USA bovine isolates) antibodies in the sera of cows; sera were further tested for antigen specificity by Western blotting. Compared to normal cows, those with digital dermatitis had a much higher seropositivity rate to B. burgdorferi and the treponemes. Significant correlations were shown between antibodies to B. burgdorferi and to Treponemes (P < 0.001), suggesting strong cross-reacting epitopes shared by these spirochaetes. In Western blotting of B. burgdorferi antigens, the main band detected by ELISA positive sera was the 41 kDa flagellar protein; lesser frequency of staining was seen with 34 (OspB), 39 and 55 kDa bands. For the USA treponeme antigens, ELISA positive sera gave reactions to the 34-kDa band and also bands at 41 and 55 kDa. Polyclonal antibodies to Treponema denticola and T. vincentii showed reactions with the bovine treponemes which were predominantly to the 34-kDa antigen. Monoclonal antibodies to B. burgdorferi flagella (41 kDa) antigen and OspA (31 kDa) did not detect any treponeme bands in Western blotting. The study has provided serological evidence that spirochaetes (which are related to human treponemes) may be involved in the pathogenesis of digital dermatitis.

The frequent detection of a treponeme in bovine digital dermatitis by immunocytochemistry and polymerase chain reaction.

A study was carried out to determine whether spirochaetes are frequently associated with digital dermatitis in United Kingdom (UK) dairy cattle. Histopathological examination of lesions using a silver stain showed a large number of unidentified spirochaete-like organisms present in digital dermatitis hoof skin tissue in all examined biopsies. Immunocytochemical staining demonstrated that spirochaetes in skin lesions were identified by polyclonal antisera to Borrelia burgdorferi, Treponema denticola and Treponema vincentii (again all biopsies were positively stained), whereas monoclonal antibodies to B. burgdorferi and any Treponema pallidum did not stain any organisms in all biopsies. A PCR of 16S rRNA, previously shown to be specific for a new treponeme, was employed and produced positive results from 82.4% of digital dermatitis tissues. It is concluded that this spirochaete (or related spirochaetes), which is similar to human oral treponemes, is frequently associated with, and may be responsible for, pathological changes in digital dermatitis.