Liver biopsy techniques for adult horses and neonatal foals to assess copper status

Objective To evaluate standing, percutaneous, ultrasound-guided, transthoracic liver biopsy in mares, and transabdominal, laparoscopically-guided, liver biopsy under general anaesthesia in foals, as techniques for obtaining tissue for assessment of copper status. The techniques were evaluated with respect to ease of use and effect on the animal.
Procedure Twenty of 24 Thoroughbred mares and 21 of their foals were biopsied. The animals were part of a larger study of the effect of copper supplementation on copper status and the prevalence of developmental orthopaedic disease. Livers were also collected from unrelated horses and sampled to investigate the variability in the distribution of copper in liver tissue.
Result The biopsy technique caused no lasting effect on the mares, but there was an increased risk of viscus penetration associated with taking multiple biopsy cores. The use of ultrasonography to scan the target area for the liver identified four cases that were not appropriate candidates for liver biopsy, because of large intestine being located in the biopsy area. In the foals there were no serious postoperative adverse effects, nor was there any evidence of problems caused by the procedure when the abdomen was examined post-mortem at 5 months of age. In livers collected to investigate the variability of copper concentration, copper appeared to be relatively evenly distributed through the liver.
Conclusion Standing, percutaneous, ultrasound-guided, transthoracic liver biopsy in mares, and transabdominal, laparoscopically-guided, liver biopsy under general anaesthesia in foals are convenient procedures for obtaining liver tissue for assessing copper status in horses. The use of ultrasound to identify liver tissue is recommended, especially in older mares.     

Macrocyclic lactone toxicity due to abamectin in farm dogs without the ABCB1 gene mutation

Abstract

CASE HISTORY: A 5-year-old entire female Huntaway from a sheep and beef farm was one of four dogs that developed clinical signs including hypersalivation, depression, blindness and ataxia after the death of another dog. A 4-year-old female Huntaway farm dog from a second farm was observed to be sitting down more often than usual on the day after being fed part of a calf carcass that had been treated with an abamectin pour-on.

CLINICAL FINDINGS: The first dog was ataxic and depressed but did respond to sound. The second dog presented with an acute onset of blindness, mydriasis, absence of a menace response, hypersalivation, gait abnormalities (e.g. high stepping gait and ataxia), and depression. Other presenting signs included muscle tremors, dehydration and difficulty eating. No abnormalities were detected from routine haematology and biochemistry. Analysis of samples of plasma from both dogs revealed concentrations of abamectin of 0.149 mg/L and 0.260 mg/L for the first and second dogs, respectively. Buccal swabs taken from the first dog for DNA testing for the ABCB1 gene mutation, gave a negative result.

DIAGNOSIS: In addition to the presenting signs which suggested a toxicosis, both dogs had measurable concentrations of abamectin in plasma confirming exposure.

CLINICAL RELEVANCE: Farm dogs exposed to concentrated pour-on products containing abamectin have been poisoned and recover or die. The product labels do not carry any warnings as to the risk of poisoning to dogs. This paper discusses two incidents affecting six farm dogs, but the authors are aware of more toxicoses in farm dogs exposed to abamectin.     

The impact of highly concentrated Mo and Cu dietary supplements,fed as a bolus,on the efficacy of chelated versus inorganic Cu in cattle on a low-Cu diet

AIM: To compare the efficacy of chelated versus inorganic forms of dietary Cu supplements, fed as a bolus, when challenged by a daily bolus of dietary Mo in cattle on a low-Cu diet.

METHODS: Forty non-lactating, Friesian dairy cows of adequate Cu status were assigned to four groups and fed a basal diet of baled silage containing 5.3 mg Cu and 0.4 mg Mo/kg DM. The experimental design was a factorial of two chemical forms of supplemental Cu and two levels of Mo intake, provided as pelleted grain supplements made from crushed barley/molasses plus Cu and Mo. The supplements contained 140 mg Cu/kg as Cu sulphate pentahydrate (CS), 140 mg Cu/kg as Cu glycinate (CG), CS plus 38 mg Mo/kg as sodium molybdate (CS+Mo), or CG plus 38 mg Mo/kg (CG+Mo). Commencing on Day 0, supplements were fed once daily (offered 1–1.2 kg/cow) and were completely consumed within 5–10 minutes, which constitutes a bolus type of administration. Liver samples were collected by biopsy at Days ?24, 13, 41 or 47, and 69 for Cu determinations.

RESULTS: The diets fed to the Cu+Mo groups were roughly equivalent to 25 mg Cu and 5.7 mg Mo/kg DM. Mean initial concentration of Cu in liver for all groups was 516 (SE 54) μmol Cu/kg fresh tissue. In cows supplemented with CS and CG, the final (Day 69) concentrations increased (p<0.01) to 939 (SE 166) and 853 (SE 163) μmol Cu/kg, respectively. These values were not different (p=0.72). For groups CS+Mo and CG+Mo, the final concentrations of 535 (SE 122) and 453 (SE 102) μmol Cu/kg were not different from initial values or from each other (p>0.25). The rate of accumulation of Cu in liver following bolus Cu and Mo intake was highly variable but was not affected by initial concentration of Cu in liver (p>0.9) or by the form of Cu (p>0.6). Mean rates of accumulation of Cu in liver were 4.0 (SD 3.8) and 0.65 (SD 2.0) μmol Cu/kg fresh tissue/day for the Cu-only treatments and the Cu+Mo treatments, respectively.

CONCLUSIONS: When fed together as a bolus, high Mo intake negated the effect of supplemental Cu but it did not reduce liver Cu stores. There was no difference in the reaction of dietary Mo with chelated Cu (as glycinate) versus inorganic Cu (as sulphate) dietary supplements.     

Ectopic pregnancy through a caesarean scar in a ewe

CASE HISTORY: A routine ultrasonographic examination for pregnancy diagnosis in a 3-year-old ewe revealed a normal embryo located in the uterine cavity and an abnormal mass located in the abdomen. At the time of examination, the ewe was 4.5?months pregnant, but 10?months previously she had undergone a caesarean section due to dystocia.

CLINICAL FINDINGS: The abnormal mass (12?cm × 8?cm) was located outside the uterine cavity; it was anechoic at the periphery, and in the centre there was an echogenic embryo-like structure. Exploratory laparotomy was performed, which revealed an ectopic pregnancy through the caesarean scar.

TREATMENT: The ectopic foetus was excised and the caesarean scar was repaired. The ewe made a complete recovery and the intrauterine pregnancy was completed to full-term, resulting in the birth of a healthy female lamb 35?days after the removal of the ectopic foetus.

CLINICAL RELEVANCE: Ectopic pregnancies have previously been described in ewes. As far as the authors are aware, this is the first report of an ectopic pregnancy through a caesarean scar in sheep or any other farm animal species.     

Impact of molybdenum on the copper status of red deer (Cervus elaphus)

AIM: To determine the effect of increasing molybdenum (Mo) intakes on serum and liver copper (Cu) concentrations and growth rates of grazing red deer (Cervus elaphus).

METHODS: Molybdenum- and Cu-amended fertilisers were applied to six 1.1-ha paddocks in a 3 × 2 design. Three levels of Mo were applied on two paddocks at each level in mid April (designated Day 1); levels were: none (control), 0.5 (medium) and 1.0 (high) kg Mo/ha as sodium molybdate. In late May (Day 39), two levels of Cu (none and 3.0 kg Cu/ha, as copper sulphate) were applied to each of the three levels of Mo-treated paddocks. Pasture Mo, Cu and sulphur (S) concentrations were measured at about fortnightly intervals. In late June (Day 74), ten 6-month-old red deer hinds were placed on the six experimental pastures, and serum and liver Cu concentrations were monitored at about monthly intervals for 102 days. The hinds were weighed on four occasions during the trial.

RESULTS: Mean pasture Mo concentrations on Day 56 were 2, 4.6 and 11.3 mg/kg dry matter (DM) for the untreated control, medium and high Mo-treated pastures, respectively. Pasture Cu concentration was 95 mg/kg DM on Day 59, 53 mg/kg DM on Day 90, and 9 mg/kg DM by Day 153. Mean S concentration in pasture was 3.3 (range 3.03–3.45) g/kg DM. Copper application to pasture had no significant effect on serum and liver Cu concentrations in deer so data were pooled within Mo treatment. Mean initial (Day 74) serum Cu concentration was 9.2 µmol/L. In the deer grazing the control Mo pasture, this increased to 10.3 µmol/L on Day 112, before decreasing to 6.4 µmol/L on Day 176. In deer grazing the medium and high Mo-treated pastures, mean serum Cu concentrations were 3.8 and 3.9 µmol/L, respectively, on Day 112, and 2.5 and 3.3 µmol/L, respectively, on Day 176. Mean initial (Day 74) liver Cu concentration was 131 µmol/kg fresh tissue. In the deer grazing the control Mo pasture, this declined to 120 and 52 µmol/kg on Days 112 and Day 176, respectively. In deer grazing the medium and high Motreated pastures, liver Cu concentrations decreased to 55 and 52 µmol/kg fresh tissue, respectively, on Day 112, and 21 and 20 µmol/kg fresh tissue, respectively, on Day 176. Mean serum and liver Cu concentrations were not significantly different between deer grazing the medium and high Mo-treated pastures, and were lower (serum p=0.003, liver p<0.001) in those groups than in deer grazing the untreated control pastures. No clinical signs of Cu deficiency associated with lameness were observed. Deer grazing pastures that had Mo concentrations >10 mg/kg DM had lower (p=0.002) growth rates (100 vs 130 g/day) than those on pastures containing <2.4 mg Mo/kg DM.

CONCLUSION: Increasing pasture Mo concentrations from 2 mg/kg DM to ≥4.6 mg/kg DM significantly reduced serum and liver Cu concentrations in grazing deer. Reduced growth rate was observed at pasture Mo concentrations >10 mg/kg DM.     

Musculoskeletal responses of 2-year-old Thoroughbred horses to early training. 1. Study design,and clinical,nutritional, radiological and histological observations

AIMS: This is the first in a series of papers reporting studies in 2-year-old Thoroughbred racehorses that aimed to determine the response of musculoskeletal tissues to early training on grass and sand racetracks. In this paper, the experimental set-up of the whole study is described, and nutritional, workload, and clinical, radiographic and pathological outcomes are reported, including semi-quantitative assessment of macroscopic changes in articular cartilage.

METHODS: The study group comprised 14 two-year-old Thoroughbred fillies reared entirely at pasture. Of these, seven were selected by a licensed racehorse trainer to undergo a 4-week period of initial training in which they were taught to accept saddle and rider, followed by a 13-week period of flatrace training at a racetrack (Weeks 1–13); the other seven fillies were confined to large grass enclosures and were not trained. Nutrient, including macro- and trace-element intakes were estimated. Distances cantered or galloped and average velocities were quantified for the trained horses. All horses were observed daily, weighed approximately weekly, and underwent a clinical lameness examination at Weeks 5, 9 and 13. Distal forelimbs were radiographed prior to Week 1, during Weeks 7–8, and again at the end of the study, when macroscopic changes in articular cartilage of the proximal surface of the proximal phalanx were also scored after staining with India ink.

RESULTS: Dietary intakes met or exceeded recommended requirements for all nutrients except sodium, which was low in the trained horses. Bodyweight increased throughout the study in the untrained horses, and increased until Week 7 and then decreased slightly in the trained horses. Mean velocity data were used to define three stages of the training programme: Stage 1 comprised canter in Weeks 1–4; Stage 2 comprised canter in Weeks 5–8; and Stage 3 comprised canter in Weeks 9–13 and galloping twice weekly. Four of seven horses completed training. These covered a mean distance of 179.2 km at mean velocities (excluding gallops) of 7.63 m/sec (SD 0.58), 8.99 m/sec (SD 0.56), and 8.43 m/sec (SD 0.74) for Stages 1–3, respectively, and galloped 4.45 km at 14.4 (SD 0.1) m/sec. The three horses that did not complete training became lame in Weeks 9, 10 and 11, and covered 147.9 km at velocities of 7.38 m/sec (SD 0.44), 8.88 m/sec (SD 0.33) and 8.43 m/sec (SD 0.59) and galloped 2.1 km. Overall, slight or intermittent lameness in trained horses was noted on 76/655 (12%) of horse observation days. Swelling was evident on 284/655 (41%) of horse observation days in the metacarpophalangeal (MCPJ) and metatarsophalangeal (MTPJ) joints (92%), palmar metacarpal tendon region (7%) or carpus (1%). Swelling of the MCPJ or MTPJ was not associated with obvious lameness. Radiographic changes were minor and no gross lesions in bone or tendon tissue were evident except for one case of dorsal metacarpal disease. Post mortem, the cartilage of some MCPJ and MTPJ had obvious wear lines and high lesion scores, which were not consistently related to clinical evidence of pain, lameness or joint swelling. Mean lesion scores were not significantly different between the MCPJ and MTPJ, or between trained and untrained horses.

CONCLUSIONS: Workload can be readily quantified in racehorses under semi-commercial training conditions. Obvious lesions in cartilage of the MCPJ or MTPJ were present in some trained and some untrained horses and not consistently associated with clinical evidence of lameness, joint swelling or change in other connective tissues.

CLINICAL RELEVANCE: Workload data in racing horses are likely to be highly relevant for studying the pathogenesis of changes in bone, tendon and cartilage during training, for training management and for risk analysis in racehorse populations. Although obvious cartilage lesions produced little clinical effect, such lesions have previously been shown to be progressive and to prejudice athletic capability. Detection of such occult lesions in young horses will require more sophisticated detection methods.     

Paleolithic population growth pulses evidenced by small animal exploitation

Variations in small game hunting along the northern and eastern rims of the Mediterranean Sea and results from predator-prey simulation modeling indicate that human population densities increased abruptly during the late Middle Paleolithic and again during the Upper and Epi-Paleolithic periods. The demographic pulses are evidenced by increasing reliance on agile, fast-reproducing partridges, hares, and rabbits at the expense of slow-reproducing but easily caught tortoises and marine shellfish and, concurrently, climate-independent size diminution in tortoises and shellfish. The results indicate that human populations of the early Middle Paleolithic were exceptionally small and highly dispersed.     

Carbamate poisoning in a dairy goat herd: Clinicopathological findings and therapeutic approach

Abstract

CASE HISTORY: Approximately 1 hour after the consumption of carnations from a nearby glasshouse 55 animals from a dairy goat herd exhibited signs of possible poisoning.

CLINICAL FINDINGS: Upon clinical examination affected animals exhibited signs of salivation, tympany, tachypnoea, polydipsia, urination, diarrhoea, bradycardia, miosis, tremor and convulsions. As poisoning from an acetylcholinesterase-inhibiting insecticide was suspected, treatment with atropine sulphate was initiated at a dose of 0.3 mg/kg bodyweight. The treatment was repeated for some animals that relapsed, and was effective in all cases, with the exception of one goat kid that died.

DIAGNOSIS: Necropsy of the goat kid showed pulmonary oedema and congestion of internal organs. Toxicological analysis of stomach contents and liver of the dead animal, as well as of the carnations, revealed high concentrations of carbamates.

CLINICAL RELEVANCE: Carbamate poisoning after consumption of polluted feedstuffs or glasshouse products can be fatal for dairy goats. Atropine sulphate, at 0.3 mg/kg, can be useful in treating this condition.