The role of Cox-2 expression in the prognosis of dogs with malignant mammary tumours

Immunohistochemical detection of Cyclooxygenase (Cox)-1 and -2 enzymes in canine mammary tumours (CMT) has recently been described. However, the prognostic value of their expression needs to be established. The aim of this study was to investigate Cox (-1 and -2) prognostic value in malignant CMT by evaluating its correlation with clinicopathological parameters (tumour size, histological type, necrosis, lymph node metastasis) and with Disease Free Survival (DFS) and Overall Survival (OS). Twenty seven female dogs with malignant tumours were included. Cox-2 expression was associated with lymph node metastasis at surgery time, development of distant metastasis during follow-up (p = 0.038), DFS (p = 0.03) and OS (p = 0.04). Multivariate survival analysis showed that Cox-2 did not retain its significance as an independent prognostic factor. For Cox-1 expression, no statistically significant association was observed. Present study suggests the usefulness of testing Cox-2 specific inhibitors as part of an adjuvant therapy in female dogs with malignant mammary neoplasias.     

PCNA and grade in 13 canine oral squamous cell carcinomas: association with prognosis

This study evaluated the prognosis factors of age, tumour size, anatomic location, histological grade and proliferating cell nuclear antigen (PCNA) expression in 13 dogs with oral squamous cell carcinoma (OSCC) with bone invasion and without signs of lymph node or distant metastasis. All animals were treated with radical excision performed with at least 1 cm margin, based on computed tomography images. In the 2‐year follow‐up, median disease‐free survival was 138 days for dogs with grade 3 tumours and was not reached for those with grade 2 tumours. Grade 3 tumours and PCNA labelling index ≥65% were related with a shorter disease‐free survival time and consequently poor prognosis (p = 0.003 and p = 0.034, respectively). Mean PCNA labelling index was significantly higher in recurrent cases (p = 0.011). Histological grade and PCNA expression may be important prognosis factors in canine OSCC.     

Timely adjuvant chemotherapy improves outcome in dogs with non-metastatic splenic hemangiosarcoma undergoing splenectomy

Timely delivery of adjuvant chemotherapy has been shown to be advantageous in many human cancers and canine osteosarcoma. Adjuvant chemotherapy has been shown to improve outcome for canine splenic hemangiosarcoma. The aim of this retrospective study was to investigate whether timely adjuvant chemotherapy administration resulted in better outcome in dogs with non-metastatic splenic hemangiosarcoma undergoing splenectomy. Medical records were searched for dogs with non-metastatic, splenic hemangiosarcoma that received splenectomy and adjuvant chemotherapy. The number of days from surgery to the first chemotherapy dose (StoC) was evaluated to identify the cut-off value associated with the best survival advantage. StoC and other possible prognostic factors were tested for influence on time to metastasis (TTM) and overall survival (OS). Seventy dogs were included. Median StoC was 20 days (range: 4–70). The time interval associated with the greatest survival benefit was 21 days. Median TTM and OS of dogs with StoC ≤ 21 days were significantly longer than those with StoC >21 days (TTM: 163 vs. 118 days, p = .001; OS: 238 vs. 146 days, p < .001). On multivariable analysis, StoC >21 days was the only variable significantly associated with increased risk of tumour progression (HR 2.1, p = .010) and death (HR 2.3; p = .008). Starting adjuvant chemotherapy within 21 days of surgery may be associated with a survival benefit in dogs with non-metastatic splenic hemangiosarcoma, possibly due to the early targeting of newly recruited metastatic cells after surgery.     

Pathophysiology of humeral fractures in a sample of dairy heifers

AIMS: To investigate the pathophysiology of humeral fractures in first-lactation dairy heifers in the North Island of New Zealand.

METHODS: Ten 2-year-old dairy heifers with humeral fractures were subject to euthanasia and the fractured and non-fractured contralateral humeri were collected. Humeri were also collected from 10 unaffected 2-year-old dairy heifers sent for slaughter. Humeri from heifers with and without fractures were examined using computed tomography (CT), and four slices of the diaphysis and lower metaphysis (D1–4) were analysed using the Bone J plug-in for Image J. The humeri were sectioned sagittally and 5?mm bone slabs were processed for histopathology.

RESULTS: There were no differences in bone length between the humeri from heifers with or without fractures (p=0.31). Median cortical bone mineral density at D1 was increased in humeri from affected compared with unaffected heifers (810 vs. 783?mg/cm³; p=0.03), cortical area at D1 was reduced (816 vs. 1,037?mm²; p=0.04), the median stress strain index, a calculated theoretical measure of bone strength, at D1 was decreased (7,288 vs. 9,072?mm³; p<0.01), and the median ratio of overall bone volume (BV) to total volume (TV) was decreased (0.32 vs. 0.38; p<0.01). The median periosteal circumference at D1 was also reduced in humeri from affected compared with unaffected heifers (151 vs. 173?mm; p<0.01). Using a binary logistic regression model, BV/TV was the only variable associated with humeral fractures (p=0.03).In nine of 10 fractured humeri the fracture appeared to have started just distal to the head of the humerus and spiralled distally down the diaphysis to end just above the humeral condyles. Histopathological findings included a reduction in the number, and thickness, of trabeculae in the metaphysis; metaphyseal growth arrest lines, and osteoclastic resorption in fractured humeri. Concentrations of copper in serum from four of five animals with fractures were within, and one was below, normal reference ranges, while concentration of copper in the livers of three heifers with fractures were below adequate ranges.

CONCLUSIONS: The CT and histological findings were consistent with a diagnosis of osteoporosis. We propose that humeral fractures in dairy heifers are associated with osteoporosis, possibly as a result of insufficient deposition of bone during growth because of protein-calorie malnutrition. Increased osteoclastic resorption of bone associated with calcium mobilisation for lactation, and periods of copper deficiency may contribute to bone weakening.     

Comparison of a novel form of zinc with zinc oxide bolus licensed for prevention of facial eczema,and interaction with copper oxide in sheep

To evaluate a novel elemental zinc bolus compared with a registered positive control zinc oxide bolus and assess serum zinc concentrations following concomitant treatment with a capsule containing copper oxide needles. Forty Romney-cross ewes were randomly allocated in a 2 × 2 factorial design study. On Day 0, 20 ewes received novel boluses containing elemental zinc (Investigational Veterinary Product, IVP) while 20 received a zinc oxide bolus (control; CP). Half the animals in each zinc treatment group (n = 10) were treated with a copper oxide needle capsule [Copasure® - Ewe]. Weekly, from Day −7 to 56, all ewes were assessed for signs of photosensitization, and for 10 ewes from each zinc treatment groups, samples were collected for analysis of serum GGT activity, serum zinc concentrations, faecal zinc concentrations and on Days −7 and 56, liver copper concentrations. Multivariable random-effects models assessed the effects of zinc treatment, copper treatment, treatment interactions and time on all analytes. Regression models examined associations between serum and faecal zinc concentrations and GGT activity. Low spore numbers indicated low Pithomyces chartarum challenge. Serum zinc levels were significantly higher in the IVP than in the CP group [p < 0.0001] and varied by time [p < 0.001] and positively associated with faecal zinc concentration [p < 0.001]. Copper treatment did not affect serum zinc [p = 0.82] or faecal zinc [p = 0.92] concentrations. Liver copper concentrations did not differ between zinc treatment groups on Day −7 [p = 0.6] or Day 56 [p = 0.95]. Only the CP/no copper group had no increase in liver copper concentrations.     

Therapeutic management of copper deficiency in buffalo heifers: Impact on immune function

To evaluate the magnitude of copper deficiency in Northern India and to examine the various haematobiochemicals, enzymes, vitamins and immune functions affected by copper deficiency, and to identify the parameters which can be of diagnostic importance in copper deficiency, a survey was conducted in 12 districts of Northern India. Significant deficiency of copper was observed in soil, fodder and serum samples of buffalo heifers. Fifty hypocuperaemic buffalo heifers were selected from these areas and were randomly divided into two groups, A and B. The heifers in group A were provided with mineral mixture containing copper sulphate and in group B without copper sulphate. Significant (p < 0.01) improvement in serum ceruloplasmin level was observed within 30 days of treatment, while significant (p < 0.01) improvement in monoamine oxidase and liver cytochrome oxidase was observed at the 60th day of treatment in group A animals. Significant improvement was observed in T₃ and T₄, in the animals of group A within 60 days of treatment. The values of vitamin A and E showed significant (p < 0.01) improvement within 30 days of treatment. The phagocytic activity of neutrophils against Candida albicans significantly (p < 0.01) improved in group A within 60 days of treatment. Similarly, significant improvement in superoxide dismutase activity in red blood cells was observed at the 30th day, and in total leukocytes and whole blood at the 60th day in group A animals. Significant improvement in liver copper level was observed at the 30th day of treatment, while in group B the liver copper was significantly (p < 0.01) depleted at the 60th day of experimentation. Additional copper supplementation improved growth performance significantly in group A.     

Erythrocyte Lipid Peroxides and Blood Zinc and Copper Concentrations in Acute Undifferentiated Diarrhoea in Calves

Undifferentiated acute calf diarrhoea is a major concern for the dairy industry and its aetiopathogenesis remains diverse. The present study aimed to examine the role of oxidative stress through estimation of erythrocyte lipid peroxide levels and blood zinc and copper concentrations using natural cases of diarrhoea in calves aged 15–30 days old. The calves were kept under identical managemental conditions and were provided with pooled whole colostrum during the first three days and thereafter with only whole milk until they were 1 month old. Diarrhoeic (n = 11) and normal calves (n = 11) of the same age group (15–30 days old) were randomly selected from an organized dairy farm for the study. The mean blood zinc concentration (50.01±2.45 μmol/L vs 66.06±3.06 μmol/L) was significantly (p < 0.05) lower and copper concentration was significantly (p < 0.05) higher in diarrhoeic calves (12.90±0.31 μmol/L vs. 9.44±0.16 μmol/L) than in the healthy calves. The erythrocyte lipid peroxides level was higher (p < 0.05) in diarrhoeic calves (6.88±0.23 nmol malondialdehyde (MDA) per mg of haemoglobin (Hb)) than healthy calves (6.27±0.07 nmol MDA per mg Hb). From the results of the study it is concluded that oxidative stress and antioxidant minerals (zinc and copper) might play important roles in the aetiopathogenesis of bovine calf diarrhoea.     

Survival analysis in dogs with urinary transitional cell carcinoma that underwent whole‐body computed tomography at diagnosis

This retrospective study aimed to evaluate factors associated with survival and to compare characteristics between tumour localizations in dogs with urinary transitional cell carcinoma (TCC) that underwent whole‐body computed tomography (CT) at diagnosis. Dogs with histologically confirmed TCC that received medical therapy between 2010 and 2017 were included; dogs that underwent surgery or radiotherapy for the primary tumour were excluded. According to the CT findings, primary tumour localization (classified into the Bladder, Urethra and Bladder and Urethra groups), prostate involvement, iliosacral lymphadenomegaly, sternal lymphadenomegaly and metastasis to the bone and lung were evaluated for survival analysis. CT at diagnosis revealed iliosacral lymphadenomegaly, sternal lymphadenomegaly, bone metastasis and lung metastasis in 47.7%, 18.5%, 24.6% and 35.4% of the 65 included dogs, respectively. The overall median survival time was 196 days. On multivariable analysis, TCC localization (hazard ratio [HR], 1.90; P = .037), bone metastasis (HR, 2.76; P = .013) and sternal lymphadenomegaly (HR, 3.56; P = .004) were significantly associated with survival. Compared to the Bladder group (n = 16), the Urethra group (n = 26) had higher metastasis rates to the bone (6.3% vs 42.3%; P = .045) and lung (6.3% vs 46.2%; P = .022). The survival time was shorter in the Urethra group than in the Bladder group (121.5 vs 420 days; P < .001), and it was similar only in female dogs (247 vs 420 days; P = .031). These findings suggest that whole‐body CT could be valuable for predicting the prognosis in urinary TCC.     

Effect of copper on the development,protein and esterase isozymes of Drosophila melanogaster

Copper is an important ingredient in fungicides, which are used for many economic plants. However, there is concern about the side‐effects of copper‐based fungicides due to their potential to affect beneficial mites. The purpose of the present paper was to investigate the effects of copper on the development of the fruit fly, Drosophila melanogaster. It was found that higher doses of copper significantly prolonged the developmental time of the fruit flies, especially during the larval stages. When 320 mg L^‐1 copper‐contaminated food was used, most of the larvae died when they were small and before the pupal stage. The protein contents and esterase isozymes extracted from the larvae changed according to the doses of copper. The small‐molecular‐weight protein bands gradually became weaker or were lost as the copper levels increased. However, low doses of copper stimulated a stronger expression of a few proteins. These results indicate that low doses of copper generally have no lethal effects on D. melanogaster because a specific group of genes, which encode specific proteins, are probably activated in order to withstand the onslaught of stressful conditions. At high doses of copper in food (e.g. 320 mg L^‐1), fly development and viability are significantly affected.     

Inter‐ and intra‐individual differences in serum copper and zinc concentrations after feeding different amounts of copper and zinc over two lactations

The influence of different copper and zinc contents in rations on blood serum concentrations was tested in a feeding trial at the Institute of Animal Nutrition of the Federal Research Institute for Animal Health (FLI). A total of 60 German Holstein breed cows (38 first lactating and 22 second lactating cows) were tested. All animals received a diet based on maize‐ and grass silage ad libitum. The animals were divided into two groups with 30 cows each. Group A received concentrates supplemented with copper and zinc as recommended, whereas Group B was offered a concentrate with roughly double the amount of copper and zinc. Blood samples were taken several times during the lactation. The concentrations of copper and zinc in the serum were measured and correlations between trace element intake and their serum concentrations were calculated. The mean dry matter intake was significantly higher for cows in the second lactation (19.1 kg/day) than for cows in the first lactation (16.4 kg/day), whereas the DM‐intake was constant in the different feeding groups (17.3 and 17.4 kg/day). The correlation between feed intake in the first and the second lactation was r = 0.76. The copper concentration in the serum was not influenced by the different feedings (11.9 and 12.5 μmol/l), but high variations were found between the cows. The influence of the lactation number was not significant (12.1 and 12.4 μmol/l). In contrast to copper, the zinc concentration in the serum was significantly higher for Group B (14.0 μmol/l) than for Group A (12.2 μmol/l), respectively, for second lactating cows (13.8 μmol/l) in relation to 12.7 μmol/l for first lactating cows. The individual variation was also high as for copper. Neither the correlation between copper intake and copper serum concentration nor the correlation between zinc intake and zinc serum concentration was significant.     

Effect of copper edetate injection in dry pregnant cows on hematology,blood metabolites,weight gain and health of calves

The objectives of the present study was to investigate the effects of the copper injection and injection timing to pregnant dairy cows at dry period on the hematology, some serum metabolites, health and growth of their calves. Thirty Holstein cows at the dry period were assigned to the three following experimental groups: 1) control (injections of 2.5 ml normal saline), 2) test 1 (125 mg Cu as copper edetate injection at the first week of the dry period) and 3) test 2 (125 mg Cu as copper edetate injection at the third week of the dry period). Ten milliliters of jugular blood was taken from all calves 24–48 h after birth and at 7, 14, 21 and 28 days of age for measuring hematological and biochemical parameters. For evaluation of growth and health, body weight of all of the calves was measured at birth and at the end of the trial, and the health of calves was checked by a technician twice a day and any sign of illness, treatment (if needed) and duration of illness was recorded. Copper treatment was proved to have a significant effect on the counts of WBC (lower in test 2, higher in test 1), lymphocyte (lower in test 2, higher in test 1) and the concentrations of fibrinogen (lower in test 1, higher in test 2), cholesterol (lower in control, higher in test 1) and copper (lower in test 1, higher in test 2) (p < 0.05). Age (sampling time) also influenced most measured parameters (p < 0.05) except values of WBC, neutrophil, monocyte, globulin and concentrations of iron, zinc and copper.     

Transcriptome sequencing reveals two subtypes of cortisol-secreting adrenocortical tumours in dogs and identifies CYP26B1 as a potential new therapeutic target

Cushing's syndrome (CS) is a serious endocrine disorder that is relatively common in dogs, but rare in humans. In ~15%–20% of cases, CS is caused by a cortisol-secreting adrenocortical tumour (csACT). To identify differentially expressed genes that can improve prognostic predictions after surgery and represent novel treatment targets, we performed RNA sequencing on csACTs (n = 48) and normal adrenal cortices (NACs; n = 10) of dogs. A gene was declared differentially expressed when the adjusted p-value was <.05 and the log₂ fold change was >2 or < −2. Between NACs and csACTs, 98 genes were differentially expressed. Based on the principal component analysis (PCA) the csACTs were separated in two groups, of which Group 1 had significantly better survival after adrenalectomy (p = .002) than Group 2. Between csACT Group G1 and Group 2, 77 genes were differentially expressed. One of these, cytochrome P450 26B1 (CYP26B1), was significantly associated with survival in both our canine csACTs and in a publicly available data set of 33 human cortisol-secreting adrenocortical carcinomas. In the validation cohort, CYP26B1 was also expressed significantly higher (p = .012) in canine csACTs compared with NACs. In future studies it would be interesting to determine whether CYP26B1 inhibitors could inhibit csACT growth in both dogs and humans.     

Doxorubicin chemotherapy for presumptive cardiac hemangiosarcoma in dogs†

Sixty‐four dogs were treated with single‐agent doxorubicin (DOX) for presumptive cardiac hemangiosarcoma (cHSA). The objective response rate (CR + PR) was 41%, and the biologic response rate (CR + PR + SD), or clinical benefit, was 68%. The median progression‐free survival (PFS) for treated dogs was 66 days. The median survival time (MST) for this group was 116 days and was significantly improved compared to a MST of 12 days for untreated control dogs (P = 0.0001). Biologic response was significantly associated with improved PFS (P < 0.0001) and OS (P < 0.0001). Univariate analysis identified larger tumour size as a variable negatively associated with PFS. The high rate of clinical benefit and improved MST suggest that DOX has activity in canine cHSA.     

Thromboelastographic results and hypercoagulability in dogs with surgically treated hepatocellular adenoma and carcinoma: A Veterinary Society of Surgical Oncology prospective study

Background

The most common haemostatic abnormality in dogs with cancer is hypercoagulability. A transient hypercoagulability has been documented in people with hepatocellular carcinoma (HCC) that resolves within weeks following hepatic tumour resection.

Objective

The objective was to compare the haemostatic status of dogs with liver tumours and healthy control dogs, by comparing coagulation and thromboelastography (TEG) measurements at three time points.

Methods

Liver tumour and healthy control dogs receiving surgery for liver lobectomy and ovariohysterectomy, respectively, were prospectively enrolled. All dogs had blood collected at three time points: pre-operative, 24 h post-operative and ~2 weeks post-operative. Haematological and haemostatic values were compared across time points in each group using repeated measures ANOVA tests.

Results

Ten and eight dogs were enrolled for the liver and control groups, respectively. Platelet count was significantly higher in the liver group than the control group at all time points, but within the normal range (pre-operative: 438.7 vs. 300.9 × 10⁹/L, p = .0078; 24 h post-operative: 416.2 vs. 283.9 × 10⁹/L, p = .0123; 10–14 days post-operative: 524.6 vs. 317.3 × 10⁹/L, p = .0072). The measure of the overall coagulant state (G-value) was significantly increased for the liver group compared to the control group at all time points (pre-operative: 15.6 vs. 8.6 d/sc, p = .0003; 24 h post-operative: 18.3 vs. 11.2 d/sc, p = .039; 10–14 days post-operative: 15.1 vs. 9.6 d/sc, p = .015).

Conclusion

The liver group was hypercoagulable based on elevated G-values at all time points compared to the control group. This hypercoagulability was attributed to the effect of hepatic tumours alone, and not secondary to surgery and anaesthesia.     

Prognostic significance of hypermethylation of death‐associated protein kinase (DAPK) gene CpG island in dogs with high‐grade B‐cell lymphoma

Death‐associated protein kinase (DAPK) is a serine/threonine kinase and a tumour suppressor gene. Diffuse large B‐cell lymphomas with inactivated DAPK through hypermethylation of a CpG island is known to result in a biologically aggressive phenotype in humans. This retrospective study was carried out to analyse the prognostic significance of DAPK CpG island hypermethylation in canine lymphoma. We hypothesized that DAPK CpG island hypermethylation can be a negative prognostic indicator in dogs with nodal high‐grade B‐cell lymphoma. Forty‐seven dogs with high‐grade B‐cell lymphoma, according to the updated Kiel classification, were evaluated after being treated with a CHOP (vincristine, cyclophosphamide, doxorubicin and prednisolone)‐based chemotherapy protocol. The methylation status of the DAPK CpG island was examined by methylation‐specific PCR. Progression‐free survival (PFS) and overall survival (OS) were compared using the Kaplan‐Meier analysis and log‐rank test. The cox proportional hazard regression model was used to evaluate the effect of multiple variables. Hypermethylation of the DAPK CpG island was detected in 21 of the 47 dogs. The PFS and OS in dogs with the hypermethylation (median: 220 and 266 days, respectively) were significantly shorter than those of dogs without hypermethylation (median: 301 and 412 days, respectively) (PFS, P = .036; OS, P = .007). In the multivariate analysis, hypermethylation of the DAPK CpG island remained an independent prognostic factor in predicting shortened PFS (P = .047) and OS (P = .021) as well as clinical substage b. Overall, hypermethylation of the DAPK CpG island was a negative prognostic factor in canine high‐grade B‐cell lymphoma.     

Acanthomatous ameloblastoma in dogs treated with intralesional bleomycin

Acanthomatous ameloblastoma (AA) is a benign gingival tumour that often invades bone. This retrospective study evaluated the efficacy of intralesional (IL) bleomycin as a treatment for AA. Six dogs received weekly or bimonthly IL bleomycin injections (dose range, 10–20 U m^?2). A seventh dog presented with advanced, nonresectable AA was treated palliatively. One to sixteen treatments were administered (median, 5). Six of the seven dogs had a complete response within 4 months from initial IL injection (median, 1.5 months), whereas the palliative case had approximately 25% decrease in tumour volume 14 days from initial injection. Local recurrence was not observed during the study period, with a median follow‐up time of 842 days. Adverse effects were limited to wound formation with bone exposure (n = 4), mild tissue reactions (n = 3), local swelling (n = 2) and local infection (n = 1). The conclusions of this study show IL bleomycin is an effective treatment for canines with AA.     

A comparison of the antinociceptive effects of xylazine, detomidine and romifidine on experimental pain in horses

Objective To study the analgesic potency of the α₂‐agonist romifidine in the horse using both an electrical current and a mechanical pressure model for nociceptive threshold testing. In addition, a comparison was made with doses of detomidine and xylazine that produce equivalent degrees of sedation. Study design Randomized, placebo‐controlled, blinded cross‐over study. Animals Six adult Swiss warmblood horses, one mare and five geldings, weighing from 530 to 650 kg and aged 6–15 years. Methods Nociceptive thresholds were measured using an electrical stimulus applied to the coronary band and using a pneumatically operated pin pressing on the cannon bone. Measurements were made immediately before and every 15 minutes for 2 hours after IV injection of the test substances. Lifting of the foot indicated the test end point. Results The three α₂‐agonists caused a temporary increase in nociceptive thresholds with a maximal effect within 15 minutes and a return to baseline levels within 1 hour. Using electrical current testing nociceptive thresholds were significantly different from placebo (mean ± SD) for detomidine at 15 minutes (from control 5.8 ± 0.9 to 23.3 ± 3.9 mA, p = 0.0066) and 30 minutes (from control 6.6 ± 1.1 to 18.8 ± 3.3 mA, p = 0.0091). The difference was significant for romifidine at 15 minutes only (from control 5.8 ± 0.9 to 18.7 ± 3.8 mA, p = 0.0066). With mechanical pressure testing nociceptive thresholds were significantly different from control for detomidine at 15 minutes (from 3.2 ± 0.2 to 6.2 ± 0.5 N, p = 0.00076) and 30 minutes (from 3.2 ± 0.7 to 5.7 ± 0.8 N, p = 0.0167). The difference was significant for xylazine at 15 minutes (from control 3.2 ± 0.2 to 5.6 ± 0.7 N, p = 0.0079). At 15 minutes the order of magnitude of the measured antinociceptive effect was significantly different between the two pain tests for both romifidine and detomidine, but not for xylazine. For romifidine, the increase of mean thresholds compared to placebo was 4.0 ± 1.3 times placebo levels with the electrical current test compared to 1.3 ± 0.3 times for the mechanical pressure test (p = 0.037). For detomidine, the increase of mean thresholds compared to placebo was 5.4 ± 1.7 times control levels with the electrical current test compared to 2.0 ± 0.2 times for the mechanical pressure test (p = 0.040). This represents a 2.7 (romifidine) and 3.4 times (detomidine) greater increase in thresholds using electrical current testing compared to the use of mechanical pressure testing. Conclusion and clinical relevance This study demonstrates the analgesic potential of α₂‐agonists in the horse for somatic pain and that they can have quantitatively different antinociceptive effects according to the antinociceptive test used.     

Effects of water‐borne copper on the gills and hepatopancreas of Macrobrachium rosenbergii

We focused on elucidating the toxic effects of water‐borne copper on the giant freshwater prawn Macrobrachium rosenbergii. After seven days of exposure to copper (Cu²⁺) at concentrations ranging from 0.01 mg L^?1 to 0.5 mg L^?1, three isozymes, malate dehydrogenase, alkaline phosphatase and esterase, were analyzed and compared using polyacrylamide electrophoresis (PAGE) and biochemical staining. The results indicated that the electrophoretic patterns of the isozymes showed a copper‐concentration‐related difference. Low doses of copper stimulated strong expression of the three isozymes. Electrophoretic patterns of malate dehydrogenase and alkaline phosphatase gradually became weaker or even lost as the level of copper increased. In contrast, esterase patterns exhibited an increased molecular heterogeneity at higher copper concentrations. A transmission electron microscope was used to study ultrastructure differences in the gills and hepatopancreas of M. rosenbergii, and the results showed significant structural damage at increased levels of copper compared with the control group. The basement membranes and mitochondira in the gills were seriously damaged, the cuticle electron density distribution was not homogeneous, and an infolded basement membrane, circularized nucleus, disintegrated nuclear membrane, and decreased mitochondria number and size were observed in the gills. Similarly, flowing out of karyoplasms, partly falling microvilli, decreased mitochondrion, partly disappeared mitochondrial cristae, and a thinned matrix were observed in the hepatopancreas. These findings indicate that exposure to elevated copper levels might damage the ultrastructure of the gills and hepatopancreas of M. rosenbergii and might further weaken their normal physical activities. Isozymes were quite sensitive to environmental stress and changes in isozyme elctrophoretic patterns might be effective biomarkers of environmental contamination.     

Antibody Response in Sheep Following Immunization with Streptococcus bovis in Different Adjuvants

Recent studies have shown that immunization with Streptococcus bovis using Freund's complete adjuvant (FCA) may confer protection against lactic acidosis in sheep. The major objective of this study was to compare the antibody responses to S. bovis in a practically acceptable adjuvant (Freund's incomplete adjuvant (FIA); QuilA; dextran sulphate (Dex); Imject Alum; or Gerbu) and in FCA. Thirty-five sheep were randomly allocated to 7 treatment groups. Six groups were immunized with S. bovis in an adjuvant; the other group served as the non-immunization control. The primary immunization was administered intramuscularly on day 0, followed by a booster injection on day 28. Immunization with FCA induced the highest saliva and serum antibody responses. The saliva antibody concentrations in the FIA and QuilA groups were significantly higher than those in the Alum, Dex and Gerbu groups (p<0.01). The serum antibody concentration in the FIA group was significantly higher than those in the QuilA, Alum, Dex and Gerbu groups (p<0.01). Immunization enhanced the antibody level in faeces (p<0.05), but there was no significant difference between the different adjuvant groups (p>0.05). Seven and 14 days following booster immunization, the saliva antibody levels induced by QuilA and/or FIA were comparable with the level stimulated by FCA (p>0.05). There was a strongly positive correlation (R ² = 0.770, p<0.01) between the antibody concentrations in saliva and serum. Compared with the controls, a higher faecal dry matter content was observed in the animals immunized with either FCA or QuilA. The change in faecal dry matter content was positively associated with the faecal antibody concentration (R ² = 0.441, p<0.05). These results indicate that FIA and QuilA were effective at inducing high levels of antibody responses to S. bovis, and suggest that either Freund's incomplete adjuvant or QuilA may be useful for preparing a practically acceptable vaccine against lactic acidosis.     

Animal fibre: The forgotten nutrient in strict carnivores? First insights in the cheetah

As wild felids are obligate carnivores, it is likely that poorly enzymatically digestible animal tissues determine hindgut fermentation, instead of plant fibre. Therefore, faecal concentrations of short‐chain fatty acids (SCFA, including branched‐chain fatty acids, BCFA), indole and phenol were evaluated in 14 captive cheetahs, fed two different diets differing in proportion of poorly enzymatically digestible animal tissue. Using a cross‐over design, the cheetahs were fed exclusively whole rabbit or supplemented beef for 1 month each. Feeding whole rabbit decreased faecal propionic (p < 0.001) and butyric (p = 0.013) acid concentrations, yet total SCFA was unaltered (p = 0.146). Also, a remarkably higher acetic acid to propionic acid ratio (p = 0.013) was present when fed whole rabbit. Total BCFA (p = 0.011) and putrefactive indole (p = 0.004) and phenol (p = 0.002) were lower when fed whole rabbit. Additionally, serum indoxyl sulphate, a toxic metabolite of indole, was analysed and showed a quadratic decrease (p = 0.050) when fed whole rabbit. The divergent SCFA ratios and the decrease in putrefaction when fed whole rabbit could be caused by the presence of undigested tissue, such as skin, bone and cartilage, that might have fibre‐like functions. The concept of animal fibre is an unexplored area of interest relevant to gastrointestinal health of captive cheetahs and likely other felids.