Copper deficiency in sheep: An assessment of the relationship between concentrations of copper in serum and plasma

AIM: To assess the relationship between concentrations of copper in serum and plasma in sheep.

METHODS: Concentrations of Cu were measured in paired serum and heparinised plasma samples collected from 110 sheep in nine flocks. Linear regression was used to evaluate whether flock or gender had a significant effect on the association between concentrations of Cu in serum and plasma. The individual results for concentrations of Cu in serum were then compared with those from plasma, using correlation and limits of agreement plotting.

RESULTS: Concentrations of Cu in serum ranged from 7.3 to 22 (mean 14.0) µmol/L, while concentrations in plasma ranged from 9 to 27 (mean 16.3) µmol/L. On average, concentrations of Cu in serum were 2.3 µmol/L lower than in plasma. Over the range of values seen in this study, concentrations of Cu in plasma and serum were significantly correlated (r=0.89), and mean concentrations in serum were 87% of those in plasma. There was no effect of flock or gender on the relationship between concentrations of Cu in serum and plasma. Despite the significant correlation, there was marked variability between individual samples in the proportion of Cu that was lost during clotting, with the 95% limits of agreement for serum Cu ranging from 70 to 104% of the plasma concentration.

CONCLUSIONS: As in cattle, the individual variability in the loss of Cu during clotting in sheep is too great for concentration of Cu in serum to be used as a substitute for that in plasma.

CLINICAL RELEVANCE: When assessing the blood Cu pool as part of the diagnosis of Cu-responsive disease in sheep, the concentration of Cu in plasma should be measured in preference to that of serum. We suggest that a range of 4.5 to 9 µmol/L in plasma be used to define marginal Cu status in sheep.     

Risk characterisation and management of sewage sludge on agricultural land-implications for the environment and the food-chain

The disposal of sewage wastes may cause severe environmental problems as was graphically demonstrated with pollution on Sydney's ocean beaches in recent years. Sewage sludges contain valuable plant nutrients and organic matter which can improve the fertility and structure of the soil. However, human parasites, pathogenic micro-organisms and chemicals capable of causing soil contamination, phytotoxicity and residues in animal products may also be present. Although sewage sludge is frequently spread on agricultural land overseas, it is not common in Australia and most states do not have specific regulations to minimise risk and promote good practice. A sludge-to-land program began in the Sydney region in 1990. It follows guidelines written by NSW Agriculture to encourage beneficial agricultural use of sludge by adoption of environmentally sustainable practices. This article describes the major risks to the food-chain and the environment, which may be associated with applying sewage sludge to agricultural land. It summarises how the risks are managed, and where further research data are required.     

Pre-hospital low-volume resuscitation with hemoglobin-glutamer-200 (Hb-200) in a canine hemorrhagic shock model

The Bowen Technique is a unique hands‐on therapy developed by Tom Bowen of Australia in the 1950s–60s and adapted for use in animals by Carol Bennett in 1997. It consists of a specific sequence of gentle, precise rolling moves done with thumb and fingers over muscle and tendon edges, across the direction of tension. A short waiting period follows each set of moves. Definitive mechanism(s) of action remains to be identified; however, autonomic nervous system modulation (heart rate variability studies) has been documented in humans. The technique has also been reported effective for back, hip, neck, and shoulder pain in human studies. Five geriatric dogs (13–16 years old) of various breeds were presented in a case series at a small animal clinic for primary complaints of stiffness, poor ambulation, and difficulty lying down/getting up. Four had hindlimb proprioceptive deficits; three had arthritic changes to coxofemoral joints and/or lumbar spine; three showed active indicators of pain (chronic pacing/panting; irritability; social withdrawal). All were on NSAID and/or nutritional therapy with unsatisfactory results. One dog had acupuncture weekly for over 1 year but no longer tolerated the needles. Treatment consisted of Animal Bowen at weekly or biweekly intervals. Three dogs were pain scored [0–10; average starting score 6.7 ± 0.6 (mean ± S.D.); range 6–7] by owner before and after treatment. All five dogs showed significant positive changes in attitude, and four had notable improvement in ease of ambulation, after the first treatment. Improvement was progressive over treatment course. Average post‐treatment score was 2.3 ± 2.1 (0, 3, 4); average pain score reduction was 4.3 ± 1.5 (range 3–6) after two to six treatments for the three dogs scored. One dog had complete resolution of hindlimb lameness of 5 years duration after three sessions. Animal Bowen, used alone or in conjunction with standard analgesics and other treatments, can be an effective therapy for chronic musculoskeletal conditions in dogs.     

Urine retention in cattle putatively associated with injection of an ivermectin and closantel anthelmintic formulation into the ischiorectal fossa

Case History: A group of 39, 19–22-month-old Friesian bulls were administered an ivermectin/closantel anthelmintic via intended S/C injection in the ischiorectal fossa on 15 June 2017 (Day 0). Over the next 50 days, 22 affected bulls presented various degrees of anorexia, abdominal pain and urine dribbling. Seventeen bulls were examined by transrectal ultrasonography which revealed urinary bladder distension in all 17, and peritoneal fluid accumulation in some. Overall, eight bulls died or were subjected to euthanasia. On-farm postmortem examination of three bulls revealed urinary bladder rupture.

Clinical Findings: On Day 50 one affected live bull was admitted to Massey University for further investigation. This bull continuously dribbled urine and had an overtly distended urinary bladder as determined by rectal palpation and ultrasonography.

Pathological Findings: Postmortem examination of this bull revealed a markedly distended urinary bladder, massive subcapsular and pericapsular renal oedema with retroperitoneal fluid accumulation, minimal hydronephrosis and no evidence of mechanical urinary outflow obstruction. The right ischiorectal fossa contained multifocal areas of tissue fibrosis that extended into areas innervated by the distal cutaneous branch of the pudendal nerve and the pelvic nerve. Histopathological changes consisted of extensive fibrosis, myonecrosis and neurodegeneration, and evidence of granulation tissue and inflammation at the putative injection site and in surrounding tissues.

Diagnosis: A local inflammatory reaction at the presumed injection site together with localised peripheral neurodegeneration and myelopathy may have led to detrusor-sphincter dyssynergia causing urine retention.

Clinical Relevance: These cases of urine retention and bladder rupture in cattle were of putative iatrogenic origin. Veterinarians should be aware of this rare complication after S/C injections in the ischiorectal fossa.     

Further evidence that zinc sulphate compromises the efficacy of dipping treatments using diazinon to control sheep lice (Bovicola ovis)

Objective To compare the wettability and efficacy of diazinon dip wash made with and without the addition of zinc sulphate.
Design Field experiments using a shower and a plunge dip complemented by in-vitro wettability experiments.
Procedure A flock of infested sheep was divided into groups and treated in a shower dip with clear or cloudy dam water plus up to 1.5% zinc sulphate. Another infested line of sheep was treated using a plunge dip with nil or 1% zinc sulphate. In both experiments, wetting was assessed after dipping and louse counts were conducted for 9 months after treatment. Five in-vitro experiments compared the wettability of dip wash containing diazinon with up to 1.5% zinc sulphate added.
Results In the shower dipping experiment, live lice were found at 1 month after dipping in the cloudy water groups with 0.75%, 1.0% and 1.5% zinc sulphate and at 2 months in the 0.75% zinc sulphate group. No lice were found at subsequent inspections or at any time in the groups that were plunge dipped. Zinc sulphate decreased the amount of dip wash retained by wool staples in all in-vitro experiments (P < 0.05).
Conclusion Zinc sulphate should be considered as a risk factor that could cause failure to eradicate a lice infestation. The risk can be overcome by ensuring that all sheep are saturated at dipping and that the dip wash, and any holding tanks, are agitated throughout the dipping event.     

Effects of local anaesthesia of 4 to 8 hours duration on the acute cortisol response to scoop dehorning in calves

Objective To determine the effect of a long-acting local anaesthetic on the plasma cortisol response in calves dehorned using a scoop.
Design A physiological study with controls.
Procedure Seventy female calves, 3 to 4 months old, were divided into seven groups and dehorned by scoop with or without a long acting local anaesthetic (bupivacaine) which provides local anaesthesia for 3 to 4 h. In some treatment groups the local anaesthetic was given a second time, after 4 h, to give 8 h of anaesthesia. Blood samples were taken before and for 9 h after treatment and plasma cortisol concentrations were measured.
Results Scoop dehorning caused a marked rise in plasma cortisol concentration, which returned to pre-dehorning values after 7 h. The cortisol concentrations of calves given bupiva-caine were similar to those of control animals for 4 h after dehorning, but then a significant and protracted rise occurred, with concentrations returning to pretreatment values at about 9 h after dehorning. In calves given a second treatment of bupivacaine 4 h after dehorning, the plasma cortisol concentration remained similar to control values for 8 h after dehorning but then increased sharply.
Conclusions During its period of activity long-acting local anaesthetic prevented the rise usually seen after scoop dehorning in plasma cortisol concentrations; by inference it also alleviated the pain-induced distress caused by this procedure. However, when its effects had worn off the plasma cortisol values rose sharply indicating that animals still experienced pain-induced distress despite local anaesthetic being administered.     

Endometrial nitric oxide synthase activity in mares susceptible or resistant to persistent breeding‐induced endometritis and the effect of a specific iNOS inhibitor in vitro

Emerging research suggests that the nitric oxide system may play a role in persistent breeding‐induced endometritis (PBIE) in the mare. Differences in uterine nitric oxide (NO) levels between mares susceptible or resistant to PBIE and a dose‐dependent inhibitory effect of NO on uterine contractility have been demonstrated. The objectives of this study were to investigate the difference in total nitric oxide synthase (NOS) activity of the endometrium between susceptible and resistant mares and the effect of a specific inducible nitric oxide synthase (iNOS) inhibitor on the endometrial NOS activity in vitro. Six susceptible and six resistant mares were selected based on preset criteria and the results of an intrauterine challenge with killed spermatozoa during oestrus. Endometrial biopsy samples were collected 24 hr post‐challenge and cultured at 37°C for 24 hr in L‐arginine supplemented minimum essential medium with or without a specific iNOS inhibitor (1,400 W dihydrochloride, 1 mM). The medium and the cultured endometrial tissue were collected after 24 hr of culture and assayed for NO and total protein, respectively. Total NO content of the medium, normalized to endometrial tissue wet weight or total protein, was used as a measure of endometrial NOS activity. Non‐parametric tests were applied for statistical analysis. Susceptible mares had significantly greater endometrial NOS activity than resistant mares. The iNOS inhibitor treatment significantly reduced NOS activity in endometrial samples derived from susceptible and resistant mares. These findings provide a basis for in vivo testing of specific iNOS inhibitors as preventative or therapeutic options for PBIE in mares.     

Identification of ectopic ovotestis in a dog with XX ovotesticular,SRY‐negative,disorder of sexual development

A 1‐year‐old, previously spayed phenotypic female Poodle/Soft‐coated Wheaten Terrier (Whoodle) cross was presented for a suspected ovarian remnant. Serum luteinizing hormone (LH) concentration was below the detection limit (<1 ng/ml Witness^® LH), and serum progesterone concentration was elevated in the chemiluminescence immunoassay (CLIA; 20 ng/ml), consistent with dioestrus and presence of ovarian tissue. Transabdominal ultrasound revealed a retroperitoneal soft tissue structure suspected to be a gonad. On exploratory laparotomy, a gonad was removed from the cranial retroperitoneum, cranial to the right kidney, after ligation of its primary blood supply. Histological examination proved the gonad to be an ovotestis. Subsequent cytogenetics revealed a 78 XX karyotype, thus confirming the diagnosis of ectopic ovotestis in a XX ovotesticular, SRY‐negative, disorder of sexual development in a dog.     

First report and histological features of Chlamydia pecorum encephalitis in calves in New Zealand

CASE HISTORY: Between September and October 2013, 40 of 150 crossbred Friesian dairy calves on a farm in the Manawatu region of New Zealand developed neurological signs when between 1 and 3 months of age. Calves were grazed in multiple mobs and calves from each mob were affected. A variable response was observed to initial treatment with thiamine, fluoroquinolone antibiotics and non-steroidal anti-inflammatory drugs.

CLINICAL AND PATHOLOGICAL FINDINGS: Affected calves exhibited a range of neurological signs that included generalised depression, hind limb ataxia with a stiff gait, and knuckling of the fetlocks. In advanced cases, calves became recumbent with opisthotonous. Over a 4-week period, 13 calves died or were subject to euthanasia and a thorough necropsy was performed on three of these calves. Necropsy findings included fibrinous peritonitis, pleuritis and pericarditis, with no gross abnormalities visible in the brain or joints. Histology of the brain was possible in seven of the affected calves, with lesions ranging from lymphocytic and histiocytic vasculitis and meningoencephalitis, to extensive thrombosis and neutrophilic inflammation. Immunohistochemistry using an anti-chlamydial lipopolysaccharide antibody revealed positive immuno-staining in all seven cases, with no brain samples exhibiting immunostaining for Histophilus somni. DNA was extracted from a sample of fresh brain from one case and chlamydial DNA sequences were amplified by PCR and found to be identical to Chlamydia pecorum. PCR was also performed on formalin-fixed brain tissue from three of the other cases, but no chlamydial DNA was amplified.

DIAGNOSIS: Chlamydia pecorum meningoencephalomyelitis (sporadic bovine encephalomyelitis).

CLINICAL RELEVANCE: This is the first time that C. pecorum has been confirmed as a cause of clinical disease in New Zealand. Practitioners should be aware of this disease as a differential in calves with neurological signs, and submit samples of formalin-fixed brain as well as fresh brain to enable confirmation of suspected cases using PCR analysis. Furthermore, these cases illustrate that the histological lesions in the brains of calves with C. pecorum are more variable than previously reported, and pathologists should be aware that histological features may overlap with those traditionally ascribed to other organisms, such as H. somni.