Influence of an n‐3 long‐chain polyunsaturated fatty acid‐enriched diet on experimentally induced synovitis in horses

Dietary n‐3 long‐chain polyunsaturated fatty acid (LCPUFA) supplementation has previously been shown to modify joint‐related inflammation in several species, although information in the horse is lacking. We investigated whether dietary supplementation with n‐3 LCPUFA would modify experimentally induced synovitis in horses. Twelve, skeletally mature, non‐pregnant mares were randomly assigned to either a control diet (CONT) or an n‐3 long‐chain fatty acid‐enriched treatment diet (N3FA) containing 40 g/day of n‐3 LCPUFA for 91 days. Blood samples taken on days 0, 30, 60 and 90, and synovial fluid collected on days 0 and 90 were processed for lipid composition. On day 91, joint inflammation was stimulated using an intra‐articular (IA) injection of 100 ng of recombinant equine IL‐1beta (reIL‐1β). Synovial fluid samples taken at post‐injection hours (PIH) 0, 4, 8 and 24 were analysed for prostaglandin E₂ (PGE₂), matrix metalloproteinase (MMP) activity and routine cytology. Synovium and articular cartilage samples collected at PIH 8 were analysed for gene expression of MMP 1 and MMP 13, interleukin‐1beta (IL‐1β), cyclooxygenase 2 (COX‐2), tumour necrosis factor‐alpha and the aggrecanases, a disintegrin and metalloprotease with thrombospondin motifs (ADAMTS)‐4 and ADAMTS‐5. A 90‐day feeding period of n‐3 LCPUFA increased serum phospholipid and synovial fluid lipid compositions of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) compared to CONT horses. The reIL‐1β injection caused an inflammatory response; however, there was no effect of dietary treatment on synovial fluid PGE2 content and MMP activity. Synovial tissue collected from N3FA horses exhibited lower expression of ADAMTS‐4 compared to CONT horses. Despite the presence of EPA and DHA in the synovial fluid of N3FA horses, dietary n‐3 LCPUFA supplementation did not modify synovial fluid biomarkers compared to CONT horses; however, the lower ADAMTS‐4 mRNA expression in N3FA synovium warrants further investigation of n‐3 LCPUFA as a joint therapy.     

Polyunsaturated fatty acids influence inflammatory markers in a cellular model for canine osteoarthritis

Although it is well recognized that dietary supplementation with fish oil improves clinical symptoms in dogs suffering from osteoarthritis, the molecular basis for the dietary benefit is not yet completely resolved in dogs. This study was designed to further clarify how polyunsaturated fatty acids (PUFA) affect key factors of cartilage degeneration in a canine cell culture system mimicking osteoarthritis. Canine chondrocytes were incubated either without or with 10 μm of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), arachidonic acid (AA) or 3.6 μm ibuprofen (Ibu) as positive control for 6 days. After the supplementation, cells were stimulated with 10 ng/ml interleukin‐1β (IL‐1β) for another 48 hr to induce osteoarthritic changes, or left unstimulated. We analysed fatty acid uptake via gas–liquid chromatography, nitric oxide (NO) production via Griess assay, prostaglandin E (PGE) production via ELISA and relative gene expression of several cartilage matrix proteinases, inducible nitric oxide synthase (iNOS) and cyclooxygenase‐2 via RT‐qPCR. After supplementation, the chondrocytes rapidly incorporated the PUFA into their fatty acid pools. The stimulation with IL‐1β caused a marked increase of most of the inflammatory markers measured. N‐3 PUFA EPA reduced IL‐induced gene expression of iNOS and corresponding production of NO. N‐6 PUFA AA also decreased iNOS and NO, but furthermore lowered gene expression of matrix metalloproteinase‐3. On the other hand, AA upregulated the aggrecanase ADAMTS‐5 and augmented the release of PGE. The effect of n‐3 PUFA DHA turned out to be negligible. Our results reveal molecular mechanisms by which PUFA affect degenerative joint disease in dogs. Of particular importance is that not only EPA but also AA decreased several inflammatory markers in our model. Thus, we conclude that an appropriate balance of both n‐3 and n‐6 fatty acids deserves more attention in dietary interventions.     

Effects of feeding a high omega‐3 fatty acids diet in dogs with naturally occurring osteoarthritis

The aim of this randomized, placebo‐controlled and double‐blinded trial was to compare the effect of a veterinary therapeutic diet (VTD) rich in omega‐3 fatty acids (omega‐3) from fish origin to a regular diet used as control (CTR) over a period of 13 weeks in dogs afflicted by naturally occurring osteoarthritis (OA). Thirty privately owned dogs were selected. Dogs had lameness confirmed by an orthopaedic examination, had stifle/hip OA and had locomotor disability based on the peak of the vertically oriented ground reaction force (PVF) measured using a force platform. At Baseline, all owners were asked to determine 2–5 activities of daily living that were the most impaired. Activities were scores (0–4) in accordance with severity using case‐specific outcome measures (CSOM). The PVF was also measured. Dogs (15/group) were then randomly assigned to receive either the CTR or the VTD. The CSOM was completed twice weekly. The recording of PVF was repeated at Week 7 and 13. The VTD‐fed dogs showed a significantly higher PVF at Week 7 (p < 0.001) and at Week 13 (p < 0.001) when compared to Baseline. From Baseline to Week 13, VTD‐fed dogs had a mean (± SD) change in PVF recording of 3.5 ± 6.8% of body weight (%BW) compared with 0.5 ± 6.1%BW (p = 0.211) in CTR‐fed dogs. This change in primary outcome was consistent with an effect size of 0.5. Conversely, dogs fed the CTR did not show significant change in PVF measurements. At the end of the study, the CSOM was significantly decreased (p = 0.047) only in VTD fed dogs. In lame OA dogs, a VTD that contains high level of omega‐3 from fish origin improved the locomotor disability and the performance in activities of daily living. Such nutritional approach appears interesting for the management of OA.     

Effects of tramadol,morphine or their combination in dogs undergoing ovariohysterectomy on peri-operative electroencephalographic responses and post-operative pain

Abstract

AIM: To compare the peri-operative electroencephalogram (EEG) responses and post-operative analgesic efficacy of pre-operative morphine or tramadol with a combination of low-dose pre-operative morphine and post-operative tramadol, in dogs undergoing ovariohysterectomy.

METHODS: Dogs undergoing routine ovariohysterectomy were treated with either pre-operative morphine (0.5 mg/kg S/C, n=8), or tramadol (3 mg/kg S/C, n=8), or pre-operative low-dose morphine (0.1 mg/kg S/C) and post-operative tramadol (3 mg/kg I/V, n=8). All dogs received routine pre-anaesthetic medication, and anaesthesia was induced with I/V thiopentone to effect and maintained with halothane in oxygen. Respiratory rate, heart rate, end-tidal halothane tension (EtHal) and end-tidal CO₂ tension (EtCO₂) were monitored throughout surgery. The EEG was recorded continuously in a three electrode montage. Median frequency (F50), total power (Ptot) and 95% spectral edge frequency (F95) of the EEG power spectra were compared during different 100-second periods of surgery: prior to and during skin incision, ligation of each ovarian pedicle, ligation of uterine body and skin closure. Post-operatively, pain was assessed using the short form of the Glasgow composite measure pain scale (CMPS-SF).

RESULTS: There was no difference in F50 or Ptot of the EEG between baseline and noxious surgical events within each treatment group, or between the three groups (p>0.05). The mean F95 was higher during the first three periods of surgery for dogs administered tramadol and low-dose morphine than those that received 0.5 mg/kg morphine (p=0.001). Dogs that received low-dose morphine and tramadol had lower CMPS-SF pain scores after ovariohysterectomy than those that received either tramadol or morphine alone (p=0.001). There was no difference in pain scores between dogs in the latter two groups.

CONCLUSION AND CLINICAL RELEVANCE: Tramadol and morphine administered pre-operatively provided an equal degree of post-operative analgesia in dogs after ovariohysterectomy. A combination of pre-operative low-dose morphine and post-operative tramadol produced better post-operative analgesia than either drug administered alone pre-operatively. Administration of analgesics pre- and post-operatively could result in improved post-operative well-being of ovariohysterectomised dogs.     

Fatty acid patterns of dog erythrocyte membranes after feeding of a fish-oil based DHA-rich supplement with a base diet low in n-3 fatty acids versus a diet containing added n-3 fatty acids

Background

In dogs, increasing the tissue n-3 fatty acid (FA) content is associated with potential benefit in some medical conditions, e.g. atopic dermatitis, cancer or heart disease. Therefore effectively and conveniently increasing tissue n-3 FA levels in dogs is of interest. Incorporation of dietary n-3 FA into cell membranes may be studied by FA analysis of erythrocyte membranes (EM), because of the correlation of its FA composition with the FA composition of other cells. Aim of the study was to determine whether an n-3 FA additive added to a control diet is as effective in increasing EM n-3 FA content as feeding an n-3 FA enriched diet. Furthermore the time course of the incorporation of dietary n-3 FA into canine EM was investigated.

Methods

Thirty dogs were randomly divided into three dietary groups with ten dogs per group. CONT got a dry dog food diet which did not contain EPA or DHA. FO got a dry dog food diet with a high EPA and DHA content. ADD got the CONT diet combined with an n-3 FA additive rich in DHA and EPA. After a feeding period of 12 weeks the additive was discontinued in ADD and these dogs were fed CONT diet for another four weeks to observe washout effects. Erythrocyte lipids were extracted from venous blood samples and their FA composition was determined by gas chromatography. The Mann-Whitney-U-test was used to detect significant differences between the different groups and time points.

Results

After one week the proportions of n-3 FA, DHA and EPA were already significantly increased in ADD and FO, apparently reaching a plateau within eight weeks. In our study DHA and not EPA was preferably incorporated into the EM. After discontinuing the administration of the additive in ADD, the n-3 FA values declined slowly without reaching baseline levels within four weeks.

Conclusions

In dogs, an increase of dietary n-3 FA content leads to a rapid inclusion of n-3 FA into EM, regardless of whether the n-3 FA are offered as an enriched diet or as a normal diet supplemented with an n-3 FA additive.     

Effects of a constant rate infusion of magnesium sulphate in healthy dogs anaesthetized with isoflurane and undergoing ovariohysterectomy

ObjectiveTo determine the effects of intravenous (IV) magnesium sulphate (MgSO₄) as a bolus followed by a constant rate infusion (CRI) on anaesthetic requirements, neuroendocrine stress response to surgery, haemostasis and postoperative analgesia in healthy dogs undergoing ovariohysterectomy.Study designBlinded randomized clinical trial.AnimalsSixteen female dogs.MethodsAfter intramuscular premedication with acepromazine (0.05 mg kg^?1) and morphine (0.3 mg kg^?1), anaesthesia was induced with diazepam (0.2 mg kg^?1) and propofol (2 mg kg^?1) intravenously and maintained with isoflurane in oxygen in all dogs. Dogs were randomly assigned to two groups, M and C. Group M received MgSO₄ (50 mg kg^?1 over 15 minutes, followed by a 15 mg kg^?1 hour^?1 CRI). Group C received an equivalent bolus and CRI of lactated Ringer's solution. In addition, all dogs received lactated Ringer's solution (10 mL kg^?1 over 15 minutes followed by 10 mL kg^?1 hour^?1). End-tidal isoflurane and carbon dioxide tensions, cardio-respiratory variables, arterial blood gases, electrolytes, ACTH and cortisol concentrations were measured at different time points. Thromboelastography (TEG) was performed pre- and post-anaesthesia. Postoperative pain was evaluated using the short form of the Glasgow Composite Pain Scale. Data were analysed with repeated measures anova and Mann–Whitney U tests (p< 0.05).ResultsNo statistically significant differences between groups were found in any of the measured variables. However, the alpha angle and maximal amplitude recorded by TEG in group M were significantly increased post-anaesthesia, but remained within the reference interval. One dog in Group M and two in Group C received rescue analgesia during recovery.Conclusions and clinical relevanceAs used in this study, MgSO₄ failed to decrease isoflurane requirements, postoperative pain and stress hormone concentrations; however, it did not produce any cardio-respiratory or major haemostatic side effects. Administration of intravenous MgSO₄ together with an opioid during ovariohysterectomy in dogs does not seem to provide any clinical advantage.     

Effect of n-3 fatty acid ratio and dose on clinical manifestations,plasma fatty acids and inflammatory mediators in dogs with pruritus

The use of n-3 fatty acids is often recommended to manage pruritus. The purpose of this study was to determine the effect of various doses of n-3 fatty acids at different n-6:n-3 ratios on plasma fatty acids, clinical response and inflammatory mediators in pruritic dogs. After baseline assessment, dogs were randomly assigned to receive diets varying in both total n-3 and n-6 fatty acid dose and n-6:n-3 ratio. The total clinical score decreased significantly in all four diet groups after 8 weeks with no difference between groups. Plasma fatty acid changes generally mirrored the fatty acid content of the test diets, although alterations appeared to depend on both the dose of n-3 fatty acids and the n-6:n-3 ratio. In this clinical trial, which controlled dietary intake of fatty acids, n-3 fatty acid supplementation did not appear to have an added benefit on clinical signs over thorough clinical management.     

Effects of a DHA‐rich unextracted microalgae as a dietary supplement on performance,carcass traits and meat fatty acid profile in growing‐finishing pigs

Two 125‐day experiments of the same design were conducted to evaluate the effects of a heterotrophically grown microalgae (AURA) containing docosahexaenoic acid (DHA) on pig performance, carcass traits and the fatty acid composition of lean and adipose tissue. In each experiment, 144 Hypor pigs were blocked by sex, allocated to three treatment groups, and fed 0, 0.25% or 0.50% AURA in isonutritive, isocaloric diets. Pigs were weighed on days 0, 28, 56, 84 and 112. Feed and water intakes were measured every 28 days. Pigs were slaughtered on day 125. Data from the two studies were analysed as a single data set. Performance and carcass traits did not differ between treatments. Both microalgae treatment levels enriched (p < .05) Longissimus lumborum (LL) and backfat in DHA and improved (p < .05) their ratios of n‐6 to n‐3 fatty acids.     

Influence of feeding a fish oil‐containing diet to mature,overweight dogs: Effects on lipid metabolites,postprandial glycaemia and body weight

The objective of this study was to determine the effect of feeding a fish oil (FO)‐containing diet on lipid and protein metabolism, postprandial glycaemia and body weight (BW) of mature, overweight dogs. Seven female dogs were randomly assigned to one of two isonitrogenous and isocaloric diets, control (CO) or FO (FO), in a crossover design. Experimental periods were 69 day, separated by a washout period of 30 day. At the beginning of the experiment, and at 30 and 60 day of feeding the experimental diets, the dogs were infused with D‐glucose (2 g/kg BW) through an intravenous catheter. Blood samples were collected for 3 hr to perform a glucose tolerance test. Nitrogen balance measurements began at 06:30 on d 63 of each experimental period and ended at 06:30 on d 69. On d 66 of each period, a single dose (7.5 mg/kg) of ¹⁵N‐glycine was administered orally for determination of protein turnover. Incremental area under the curve and glucose concentration at peak did not differ between treatments or among sampling days within treatment. Glucose half‐life tended to decrease (p < .10) in the FO treatment on day 30 when compared to baseline (day 0). β‐hydroxybutyrate, non‐esterified fatty acid (NEFA) and triglycerides did not differ within or between treatments. Cholesterol decreased (p < .05) on the FO treatment on day 30, 60 and 69 when compared to day 0. High‐density lipoprotein (HDL) decreased (p < .05) in the FO treatment on day 69 when compared to day 0. Body weight, food intake, faecal excretion, DM and N digestibilities, N balance and protein turnover were not different between diets. Overall, FO‐containing diet decreases cholesterol in mature overweight dogs; however, further research is warranted to verify the effects of FO on glucose metabolism.     

Effects of dietary fish oil and alpha-tocopherol supplementation on selected blood parameters and fatty acid profiles in mares and their foals

The effects of fish oil (40 ml/day) supplementation, with or without synthetic all-rac-alpha-tocopherol-acetate (2,500 IU/day), during the last 65 days before expected parturition were investigated in 15 adult mares (553 ± 24 kg BW) and their foals. Mares were assigned to one of three diets: control (n = 5), control plus fish oil and alpha-tocopherol (n = 4; FO + AT) or control with just fish oil (n = 6; FO). Blood samples were obtained from the mares before a 15-day dietary adaptation period (T1) and from mares and foals the first (T2) and fifth (T3) days post-partum. Colostrum was collected at T2 and milk at T3. Routine haematological, biochemical and alpha-tocopherol analyses were undertaken on all blood samples. Fatty acid concentrations were determined in the foal serum and alpha-tocopherol concentrations measured in the milk and colostrum. Diet had no effect on haematology or biochemistry in the mares. Alpha-tocopherol concentrations were significantly higher at T2 & T3 in the FO + AT mares. Foal WBCs were higher in FO (11.33 ± 2.59 × 10⁹/l), comparing to FO + AT and control groups (9.18 ± 1.24 × 10⁹/l and 7.26 ± 1.03 × 10⁹/l, respectively), at T3 (p < .05). There was no significant effect of the fish oil supplementation on the foal's serum fatty acid profile. In the FO + AT group, both colostrum and milk alpha-tocopherol concentrations (2.56 ± 0.36 and 1.36 ± 0.22 µg/ml, respectively) were higher compared than those of the FO group (1.33 ± 0.39 and 0.72 ± 0.31 µg/ml, respectively; p < .05). Additional 2,500 IU/day of synthetic alpha-tocopherol in the last 65 days of pregnancy increased alpha-tocopherol concentrations in colostrum and milk and the foal's serum. 40 ml/day fish oil, however, did not significantly increase serum eicosapentaenoic acid and docosahexaenoic acid concentrations in the foals.     

Microalgae Schizochytrium sp. as a source of docosahexaenoic acid (DHA): Effects on diet digestibility,oxidation and palatability and on immunity and inflammatory indices in dogs

The objective of this study was to evaluate the effects of the microalgae Schizochytrium sp., as a dietary source of docosahexaenoic acid (DHA), on diet palatability, coefficients of total tract apparent digestibility (CTTAD) of nutrients and metabolizable energy (ME), blood variables and indicators of immunity in dogs. We also evaluated oxidative stability. Two diets containing 0 and 0.4% of microalgae Schizochytrium sp. were evaluated in three experiments. On Experiment I the palatability of diets containing 0% versus 0.4% microalgae was compared. In Experiment II test diets were offered for 30 days to determine digestibility, fecal characteristics, and blood parameters. In Experiment III, the oxidative stability of diets containing microalgae versus anchovy oil was evaluated. There was a higher intake ratio of the diet containing microalgae (p < .05). The ME and CTTAD of nutrients increased (p < .05), except for ether extract after acid hydrolysis, with the inclusion of the microalgae in diet. The amount of monocytes and phagocytic granulocytes was higher (p < .05) in dogs fed 0.4% microalgae. There was greater oxidative stability for the sample containing microalgae. The addition of 0.4% microalgae presented high palatability, increased phagocytic cell numbers, and demonstrated oxidative stability superior to anchovy oil.     

Diaphragmatic dysfunction in dogs with cervical spinal disorders before and after surgery using fluoroscopy,motion‐mode ultrasound and radiography was not different than a group of control dogs

Cervical spinal disorders can lead to life‐threatening respiratory complications. Diaphragmatic dysfunction is attributed to spinal cord morbidity secondary to cervical myelopathy or decompressive surgical intervention. The purpose of this observational case‐control study was to determine the frequency of diaphragmatic dysfunction in dogs with cervical spinal disorders and a control group, the strength of association between cervical myelopathies and decompressive surgery with diaphragm paresis, and the agreement between and clinical usefulness of fluoroscopy, motion‐mode ultrasonography, and radiography for diagnosing diaphragmatic dysfunction. Thirty‐five client‐owned dogs were recruited with 14 control dogs and 21 test dogs. Dogs were evaluated for the presence of diaphragmatic dysfunction using radiography, M‐mode ultrasonography, and fluoroscopy before and after an anesthetic or surgical event. Diaphragmatic dysfunction was observed more frequently in dogs with cervical spinal disease prior to surgery (8/21; 38.1%) compared to control dogs (3/14; 21.4%) but was not statistically significant (P = .30). The occurrence of diaphragmatic dysfunction did not significantly increase following surgical decompression in either group. There was no to slight agreement between all imaging modalities. Most dogs with diaphragmatic dysfunction were asymptomatic. Diaphragmatic dysfunction was not statistically associated with cervical myelopathy or decompressive surgery. Ultrasonography and radiography were not useful diagnostic tests for determining diaphragmatic dysfunction in asymptomatic dogs when compared to fluoroscopy.     

丹参多糖对免疫性肝损伤小鼠肝脏过氧化指标、炎症细胞因子和ICAM-1的影响

将SPF级雄性昆明小鼠随机分成正常对照组,LPS模型组,丹参多糖低、中、高剂量组和阳性药物对照组。灌胃相应药物12 d后,腹腔注射LPS和D-GalN建立小鼠免疫性肝损伤模型。检测各组小鼠肝组织中SOD、MDA、GSH-Px水平,比较造模1,3 h后各组小鼠肝组织中IL-1β、IL-6、TNF-α的含量变化以及测定肝组织ICAM-1蛋白的表达水平。结果显示,与正常对照组比较,LPS模型组小鼠肝组织中MDA含量显著升高,SOD、GSH-Px活性显著降低, IL-1β、IL-6、TNF-α含量以及ICAM-1蛋白表达水平均显著升高(P<0.01);与LPS模型组相比,丹参多糖各剂量组和阳性药物对照组小鼠肝组织中MDA含量显著降低,SOD、GSH-Px活性显著升高,IL-1β、IL-6、TNF-α含量以及ICAM-1蛋白表达水平均显著降低(P<0.05或P<0.01);各用药组之间相比较,丹参多糖中、高剂量组和阳性药物对照组调控过氧化指标、炎症细胞因子和ICAM-1效果最显著(P<0.05或P<0.01)。结果表明,丹参多糖具有明显缓解小鼠免疫性肝损伤的作用,其机制可能与拮抗肝脏脂质过氧化反应,降低肝组织中炎症细胞因子含量及ICAM-1蛋白表达有关。     

鱼油对仔猪生产性能、炎性介质和下丘脑-垂体-肾上腺轴激素的影响

本试验研究了鱼油对断奶仔猪生产性能、炎性介质和下丘脑-垂体-肾上腺轴激素的影响.选用32头(28±3)日龄、体重(8.91±0.74)kg的杜洛克×长白×大白仔猪,随机分为4组,每组4个重复,每个重复2头猪,1公1母.采用双因子设计,主因子包括:1)饲粮处理(5%鱼油或5%玉米油);2)免疫应激[注射脂多糖(LPS)或...