Quantification of mucosa-adhered microbiota of lambs and calves by the use of culture methods and fluorescent in situ hybridization coupled with flow cytometry techniques

The intestinal mucosa-associated microbiota could play important biological roles due to its close proximity with the animal host, but knowledge on its composition is still limited. The aim of this study was to characterize the microbial communities tightly associated with different parts (rumen, duodenum and colon) of the gastrointestinal tract (GIT) of healthy lambs and calves by using both cultural, and fluorescent in situ hybridization-flow cytometry (FCM-FISH) techniques. Lactic acid bacteria genera were one of the predominant bacteria detected in lambs and calves by both methodologies, possibly constituting an index of their healthy status. The levels of Lactobacillus were significantly higher (p<0.05) in the rumen and duodenum of lambs, and in the rumen of calves. The levels of Bifidobacterium were significantly higher (p<0.05) in the colon of both animal species and the rumen of lambs. Significant differences (p<0.05) were found in counts of other microbial groups (yeast, Enterococcus, Propionibacterium, Bacteroides, Clostridium and Enterobacteriaceae) at diverse GI sections depending on the animal species. In general, microbial counts follow the same trends regardless the applied technique. The most remarkable differences were found in detection levels of Bacteroides and Clostridium, which tended to be significantly higher (p<0.05) when analysed by FCM-FISH. This technique also allowed the detection of quantitatively important bacteria (sulphate-reducing bacteria, Atopobium and Coriobacterium), which are difficult to cultivate in selective medium. Therefore, FCM-FISH has been proven to be a sensitive high throughput approach that provides additional information to that obtained by traditional culture techniques about the complexity of the GI ecosystem of these animal species.     

Journey length and high temperatures: effects on rabbit welfare and meat quality

The transport of domestic animals by road can increase levels of stress and decrease meat quality, especially in unfavourable climates. The purpose of this study was to investigate the effect of journey duration and vertical position on the transport truck on some physiological indicators of stress and on instrumental meat quality parameters in commercial rabbits. In the summer months (June and July, 2003), 78 rabbits were subjected to either long (7 h) (LJ) or short (1 h) (SJ) journeys (3 replicates each, n = 6) between the farm and an abattoir in northern Spain. The position (top, middle, or bottom) occupied by the rabbits on the Multi-Floor cage Rolling Stand (MFRS) of the transport truck was recorded. Blood samples were collected at sticking and meat pH was measured at 24 h post-mortem (pH24). At 48 h post-mortem, samples of the M. longissimus dorsi were used to determine water-holding capacity (WHC) and instrumental tenderness using an INSTRON machine. The levels of corticosterone, glucose, lactate, and creatine kinase were slightly higher in LJ than in SJ samples, but the difference was not statistically significant (p < or = 0.10). Independent of journey length, rabbits in the middle and bottom of the MFRS showed higher levels (p < or = 0.05) of glucose and creatine kinase (middle), and corticosterone (bottom) than the rabbits located at the top floor. The pH 24 and WHC values of the SJ and LJ rabbits did not differ significantly. Journey length had a significant effect (p < or = 0.01) on meat tenderness. The meat from rabbits subjected to LJ had higher values of maximum stress and total toughness than did the SJ rabbits (p < or = 0.05). There were similar differences (p < or = 0.05) between LJ and SJ rabbits in their stress values of compression at 20% and 40% (measures of the tenderness of raw meat). In hot weather, the position on the transport truck appeared to have a greater effect on rabbit welfare than the duration of the journey.     

Histomorphometry of fragmented medial coronoid process in dogs: a comparison of affected and normal coronoid processes

OBJECTIVE: To determine if fatigue microdamage is associated with fragmented medial coronoid process (FMCP). STUDY DESIGN: Prospective study. ANIMALS: Thirty-eight dogs were admitted for subtotal coronoid ostectomy as treatment for FMCP. Surgical specimens of medial coronoid process (MCP) were retrieved from treated dogs. Normal MCP were collected from 5 age- and breed-matched dogs and 2 age-matched research beagles that were euthanatized for reasons other than elbow disease. METHODS: MCP specimens from affected elbows were classified into 4 groups based on disease severity. Specimens were bulk-stained for microdamage using 1% basic fuchsin, embedded in polymethylmethacrylate and sectioned at 130 microm. Specimens were evaluated using epifluorescent microscopy (425-440 excitation, 475 nm barrier filter) for diffuse damage (D.DX), osteocyte loss, and porosity. Cartilage thickness was evaluated using bright field microscopy. Qualitative observations were also made. RESULTS: MCP specimens from affected elbows had increased D.Dx, increased osteocyte loss, and greater porosity than normal groups. Specimens classified as severe FMCP (Classes 3, 4) had the most fatigue microdamage. Some Class 1 specimens, defined as having no visible fissuring of the articular cartilage (AC), had large microcracks in trabecular bone. CONCLUSIONS: Accumulation of subchondral fatigue microdamage, is associated with MCP fragmentation. Fissuring occurs in the subchondral bone before gross fibrillation of articular cartilage develops. CLINICAL RELEVANCE: Accumulation of fatigue microdamage in the MCP is important in the pathogenesis of FMCP. The underlying cause of this microdamage accumulation must be identified before treatment plans that will prevent further osteoarthritis of the elbow joint can be designed.     

Femoral bone adaptation to unstable long-term cemented total hip arthroplasty in dogs

OBJECTIVE: To evaluate femoral adaptation after unstable long-term cemented total hip arthroplasty (cTHA) in dogs. STUDY DESIGN: Clinical study. ANIMALS: Four dogs. METHODS: Paired femurs were examined from client-owned dogs that were donated to a retrieval program after death from causes unrelated to their cTHA. Mean (+/-SEM) dog age was 10.0+/-1.5 years and implant duration was 6.0+/-1.4 years. Implanted femurs had radiographic changes associated with implant loosening and gross mechanical instability at femur retrieval. Femurs were evaluated at 3 levels relative to implant length. Cortical area and medullary area were measured at each level, and cortical porosity was assessed at each level in 4 quadrants and in 3 regions. Implanted femurs were compared with the contralateral non-implanted femurs. RESULTS: Cortical area and cortical porosity were increased at all levels in femurs with unstable implants. Implanted femurs had increased porosity in all quadrants and regions at the proximal 2 levels, and increased porosity in only the cranial quadrant and mid-cortical area at the distal level, when compared with contralateral non-implanted femurs. Corresponding medullary areas were not different. CONCLUSIONS: Significant histomorphometric changes occur in femurs after unstable cTHA. The patterns of periosteal bone formation and endosteal bone resorption support mechanisms of stress shielding and wear debris-mediated osteolysis as factors that may contribute to femoral adaptation and implant loosening. CLINICAL RELEVANCE: Despite popularity and excellent return to function with cTHA, aseptic loosening remains a serious long-term complication. Substantial net bone loss and unfavorable environment with unstable cTHA may make revision surgery less successful.     

RESEARCH PAPER: Segmental dorsolumbar epidural analgesia via the caudal approach using multiple port catheters with ketamine or lidocaine or in combination in cattle

ObjectiveTo determine the analgesic and systemic effects of epidural administration of ketamine, lidocaine or a combination of ketamine/lidocaine in standing cattle.Study designProspective, randomized, experimental trial.AnimalsSix healthy male cattle weighing between 335 and 373 kg.MethodsThe animals received 0.5 mg kg^?1 of ketamine (K), 0.2 mg kg^?1 of 2% lidocaine (L) or 0.25 mg kg^?1 ketamine plus 0.1 mg kg^?1 lidocaine (KL). All the drugs were injected into the dorsolumbar epidural space via a caudal approach through a non‐styletted multiple‐port catheter. Each animal received each treatment at random. Evaluations of analgesia, sedation, ataxia, heart rate, arterial pressure, respiratory rate, skin temperature and rectal temperature were obtained at 0 (basal), 5, 10, 15, 30, 45, 60, 75, 90 minutes after epidural injection, and then at 30‐minute intervals until loss of analgesia occurred. Skin temperature was taken at these intervals up to 60 minutes. All the animals received a standard noxious stimulus; a 4‐point scale was used to score the response. A second scale was used to score ataxia and a third for sedation.ResultsThe duration of analgesia in the upper and lower flanks in cattle was 140 ± 15, 50 ± 14 and 80 ± 22 minutes (mean ± SD) after dorsolumbar epidural KL, K or L, respectively. The cardiovascular changes were within acceptable limits in these clinically healthy cattle.ConclusionsDorsolumbar epidural administration of KL to cattle resulted in longer duration of analgesia of the upper and lower flanks in standing conscious cattle, than the administration of K or L alone.Clinical relevanceFurther research is necessary to determine whether this combination using this technique provides sufficient analgesia for flank surgery in standing cattle.     

Evaluation of an antigen capture ELISA for the early diagnosis of Hypoderma lineatum in cattle under field conditions

An antigen capture or sandwich ELISA (sELISA) was evaluated for the diagnosis of Hypoderma lineatum in cattle under field conditions in northwestern Spain. The kinetics of circulating hypodermin C (HyC) and specific antibodies during the course of natural infestation were determined in a group of 10 Frisian calves. In addition, oesophagi and blood samples were taken from 105 cows at a slaughterhouse in order to compare three methods for the diagnosis of H. lineatum: sandwich ELISA for the detection of the antigen HyC (sELISA), indirect ELISA for the detection of antibodies anti-HyC (iELISA) and the detection of first instars (L1) in the oesophagus. In naturally infested cattle, HyC was present in circulation at low levels during the early and late phases of the infestation. However, in the middle phase, coinciding with the presence of L1 in the oesophagus, two peaks of increased HyC concentration were observed. Specific antibodies increased progressively until the first appearance of larvae in warbles on the back. There was no correlation between antigen or antibody levels and the number of grubs in the back. Prevalence of first instars in the oesophagi of slaughtered cows was 21.9% (23/105). The percentage of cattle that were positive for circulating antigen was slightly higher (24.8%), suggesting the recent destruction of migrating larvae in some animals. However, there was no correlation between the number of L1 and HyC levels. With the iELISA, 79% of the animals were positive to Hypoderma, which means that a high percentage of those animals have been exposed to the parasite but they had no apparent current infestation. The sELISA is a good tool to follow larval development within the host; however, the episodic elevation of HyC levels limits the usefulness of this test for the early diagnosis of Hypoderma under field conditions.     

Comparison of two automated spectrophotometric methods for ceruloplasmin measurement in pigs

Ceruloplasmin (Cp) determination could provide an objective measure of the health status of an animal and could be used as marker of animal health and welfare (Skinner, 2001) (Skinner, J. G., 2001. Special report. International standardization of acute phase proteins. Veterinary Clinical Pathology 30, 2-7.) but only manual methods have been reported to determine Cp concentrations in pigs (McCosker, 1961; Toussaint et al., 1995; Eckersall et al., 1996) (McCosker, P. J., 1961. Paraphenylenediamine oxidase activity and copper-levels in mammalian plasmas. Nature. 190, 887-889; Toussaint, M. J. M., Van Ederen, A. M., Gruys, E., 1995. Implication of clinical pathology in assessment of animal health and in animal production and meat inspection. Comparative Haematology International 5, 149-157; Eckersall, P. D., Saini, P. K., McComb, C., 1996. The acute phase response of acid soluble glycoprotein, alpha(1)-acid glycoprotein, ceruloplasmin, haptoglobin and C-reactive protein, in the pig. Veterinary Immunology and Immunopathology 51, 377-385). In the present study two automated methods based on the use of two different substrates for the determination of serum ceruloplasmin in pigs were developed, evaluated and compared. Both methods showed a good precision and detection limits, with no signs of inaccuracy and could be applied to biochemical autoanalyzers usually found in clinical laboratories using only minimal amounts of serum. Additionally the behaviour of Cp in experimental and clinical situations was studied showing an increase of around two fold after turpentine administration and significantly higher values in cases of pigs with inflammatory conditions when compared with healthy pigs.