Role of anthranilic acid in background levels of sulfonamide in porcine livers when determined by the Tishler method

Anthranilic acid occurs in excised swine livers as a result of temperature abuse before and/or after freezing. The tryptophan metabolite is the major source of the variable non-sulfonamide background level frequently encountered in the Tishler method for determining sulfonamide residues in swine livers. Diazotized anthranilic acid reacts slowly with N-1-(naphthyl)ethylenediamine [k(s-1) = 5.79 (+/- 0.07) X 10(-5)] and the final background level depends on the time elapsed between addition of the coupling agent and spectrophotometric determination. Kynurenine was tentatively identified as a minor source of the non-sulfonamide background level.     

Distribution of blood flow during moderate and strenuous exercise in ponies (Equus caballus)

Blood flow to the brain, heart, kidneys, diaphragm, and skeletal muscles was studied at rest and during graded treadmill exercise, using radionuclide-labeled microspheres (15 microns diameter), in 11 healthy adult ponies. Hemodynamic changes brought about by exercise included marked increases in cardiac output, mean aortic pressure, left ventricular end-diastolic pressure, and right ventricular systolic and end-diastolic pressures. Blood flow to the brain stem and cerebral hemispheres was unchanged during both moderate exercise (heart rate = 154 +/- 3 beats/min) and severe exercise (heart rate = 225 +/- 7 beats/min). Despite marked hypocapnia during severe exercise, cerebellar blood flow increased by 32% above control value (94 +/- 7 ml/min/100 g). Myocardial blood flow increased transmurally with both levels of exercise. The endo:epi (inner:outer) perfusion ratio for the left ventricle and the interventricular septum decreased during exercise. It was, however, not different from unity. During severe exercise, renal blood flow decreased to 19% of its control value. Blood flow to the diaphragm exceeded that to the skeletal muscles during both intensities of exercise. Blood flow to the exercising muscles of the brachium and thigh increased by 31- to 38-fold during moderate exercise and by 70- to 76-fold during severe exercise. It is concluded that the cardiovascular response to strenuous exercise in the pony included an increase in blood flow to the cerebellum, myocardium, diaphragm, and exercising skeletal muscles, while blood flow was diverted away from the kidneys. It would appear that the pony's cardiovascular response to severe exercise is similar to that of persons.     

Gut reactions of radioactive nitrite after intratracheal administration in mice

Intratracheal administration to mice of radioactive nitrite labeled with nitrogen-13 (13NO2-) (half-life, 9.96 minutes) in dosages that do not cause pharmacological perturbation reveals that oxidative and reductive reactions occur in different organs. Oxidation of 13NO2- to radioactive nitrate (13NO3-) predominates in the blood and liver. Reduction of 13NO2- occurs in those mice that harbor intestinal microflora; this reduction does not occur in germ-free mice. The intestinal reduction products include ammonium, glutamate, glutamine, and urea. With a detection limit of about 0.01 percent of the instilled nitrogen-13, no labeled nitrosamines were detected within 30 minutes. Reduced nitrogen-13 is transported out of the intensive into the circulatory system and appears in the urine along with 13NO3-. The biological half-period for 13NO2- destruction is about 7 minutes, and both oxidation and reduction products are formed.     

A non-invasive technique for standing surgical repair of urinary bladder rupture in a post-partum mare: a case report

An 11-year-old mare presented 36 hours after foaling with a ruptured bladder. Uroperitoneum was diagnosed on ultrasound and from the creatinine concentration of the peritoneal fluid. Bladder endoscopy demonstrated tissue necrosis and a rent in the dorsocranial aspect of the bladder. Following stabilisation, including abdominal drainage and lavage, the mare was taken to standing surgery. Under continuous sedation and epidural anaesthesia, and after surgical preparation, a Balfour retractor was placed in the vagina. Using sterile lubricant and moderate force, it was possible to insert a hand into the bladder. The tear was easily palpable on the dorsal portion of the bladder. Two fingers were inserted through the tear and used to provide traction to evert the bladder completely into the vagina where it could grasped with the surgeons other hand to prevent further trauma. A second surgeon could then visualise the entire tear and repaired this using a single layer of size zero PDS suture in a single continuous pattern. As soon as the bladder was repaired, it was replaced via the urethra. The mare did well after surgery and was discharged after 48 hours, apparently normal.This report is the first describing repair of the bladder without an abdominal incision or incision into the urethral sphincter. This greatly reduces the chance of possible complications such as urine pooling after surgery with the previously described standing technique or bladder trauma due to traction with abdominal surgery.     

Shedding patterns of dairy calves experimentally infected with Mycobacterium avium subspecies paratuberculosis

Although substantial fecal shedding is expected to start years after initial infection with Mycobacterium avium subspecies paratuberculosis (MAP), the potential for shedding by calves and therefore calf-to-calf transmission is underestimated in current Johne’s disease (JD) control programs. Shedding patterns were determined in this study in experimentally infected calves. Fifty calves were challenged at 2 weeks or at 3, 6, 9 or 12 months of age (6 calves served as a control group). In each age group, 5 calves were inoculated with a low and 5 with a high dose of MAP. Fecal culture was performed monthly until necropsy at 17 months of age. Overall, 61% of inoculated calves, representing all age and dose groups, shed MAP in their feces at least once during the follow-up period. Although most calves shed sporadically, 4 calves in the 2-week and 3-month high dose groups shed at every sampling. In general, shedding peaked 2 months after inoculation. Calves inoculated at 2 weeks or 3 months with a high dose of MAP shed more frequently than those inoculated with a low dose. Calves shedding frequently had more culture-positive tissue locations and more severe gross and histological lesions at necropsy. In conclusion, calves inoculated up to 1 year of age shed MAP in their feces shortly after inoculation. Consequently, there is potential for MAP transfer between calves (especially if they are group housed) and therefore, JD control programs should consider young calves as a source of infection.     

Improvement in In Vitro Fertilization Rate,Decrease in Reactive Oxygen Species and Spermatozoa Death Incidence in Rams by Dietary Fish Oil

Our aim was to evaluate the effects of fish oil feeding on sperm classical parameters, level of reactive oxygen spices (ROS), spermatozoa death incidence and in vitro fertilization (IVF) rate in rams. We randomly assigned nine rams, into two experimental groups (isoenergetic and isonitrogenous rations with constant level of vitamin E supplement): control (CTR; n = 5) and fish oil (FO; n = 4, 35 g/day/ram). Diets were fed for 70 days during the physiological breeding season. After a 21‐day dietary adaptation period, semen was collected weekly from each ram by an artificial vagina. Sperm classical parameters were determined by the computer‐assisted sperm analyzer system (CASA), and it was prepared for IVF process by swim‐up technique. These evaluations were performed during the first and last weeks of sampling. Intracellular ROS level and spermatozoa death incidence were detected by flow cytometry on a weekly basis after adaptation. Data were analysed with SPSS 15. The volume, concentration (3.6 and 2.7 × 10⁹/ml) and sperm progressive motility (60 and 48%) were significantly improved in the FO group compared with the CTR (p < 0.05). A comparison of two‐cell stage embryos following IVF in the two groups showed a significantly higher fertilization rate in the FO group (56%) compared with the CTR (49%). Superoxide anion (O₂^?) rate was significantly lower (p < 0.05) at the third week of sampling in the FO. Although the H₂O₂ rate was numerically lower in the FO group compared with the CTR, this difference was not significant. In addition, apoptosis showed a significant difference in the third week of sampling (15 and 30% for FO and CTR, respectively; p < 0.05). Overall, adding fish oil to the ram diet not only improved sperm quality and IVF results, it also could reduce oxygen‐free radicals and the incidence of spermatozoa death.     

Loss of reserves of Cu in liver when Cu supplements are withdrawn from dairy herds in the Waikato region

Abstract

AIM: To monitor the consequences of withdrawing mineral Cu supplements from two dairy herds with initially high concentrations of Cu in liver.

METHODS: Two herds were selected from dairy farms in the Waikato region of New Zealand that participated in an earlier survey of Cu supplementation practices and Cu status of dairy cows. The herds were fed pasture, grass and maize silage, plus palm kernel expeller (PKE) containing 25–30 mg Cu/kg dry matter (DM) fed at 2–4 kg/cow/day. No mineral Cu supplements were supplied from January 2009. Pasture samples were collected for mineral analysis in September 2008 and April 2009. Concentration of Cu in liver biopsies from the same 9–10 cows per herd was measured on three occasions between April 2009 and May 2010.

RESULTS: Pastures on both farms contained 10 mg Cu/kg DM, 0.1–0.5 mg Mo/kg DM and 3.5–4.0 g S/kg DM. The initial herd mean concentrations of Cu in liver were 1,500 (SD 590) and 1,250 (SD 640) μmol Cu/kg fresh tissue. In the absence of mineral Cu supplements, those mean concentrations decreased over 12 months to 705 (SD 370) and 1,120 (SD 560) μmol Cu/kg fresh tissue, respectively. For cows in the first herd, the rate of depletion of liver Cu reserves was influenced by initial concentration of Cu, such that high concentration led to faster loss according to first-order kinetics.

CONCLUSIONS: Mineral Cu supplementation was not necessary over 12 months for two dairy herds with mean concentrations of Cu in liver >1,250 μmol Cu/kg fresh tissue, grazing pastures containing 10 mg Cu/kg DM and concentrations of Mo <1 mg/kg DM. The quantity and particularly the duration of feeding PKE appeared to be a factor in whether or not the herd lost substantial reserves of Cu in liver during the year. However, the Cu status of both herds in this study was more than adequate to support late pregnancy and mating.

CLINICAL REVELANCE: Copper status of the herd should be monitored and on-farm management of Cu nutrition should take into account all sources contributing to daily intake of Cu. Where Cu supplementation has been excessive and there is risk of chronic Cu toxicity, mineral Cu supplements may be withdrawn for a period commensurate with the expected rate of liver Cu depletion.     

A critical evaluation of serum methylmalonic acid and vitamin B12 for the assessment of cobalt deficiency of growing lambs in New Zealand

AIM: To derive reference ranges for serum methylmalonic acid (MMA) for the diagnosis of cobalt/vitamin B₁₂-responsiveness in lambs and critique existing serum vitamin B₁₂ reference ranges.

METHODS: Individual animal data from earlier supplementation trials, involving 225 ewes, 106 suckling lambs, 301 lambs during the suckling and post-weaning periods and 414 weaned lambs, for which weight gain to supplementation was observed, were used to derive relationships between serum vitamin B₁₂ and MMA, and liveweight gain.

RESULTS: Serum MMA concentrations were rarely elevated above the norm of <2 µmol/L when serum vitamin B₁₂ concentrations were >375 pmol/L, and not elevated into the range where a liveweight response to supplementation occurred (>10 µmol/L) unless serum vitamin B₁₂ concentrations were below 200 pmol/L. Suckling lambs were able to maintain high growth rates despite elevated serum MMA concentrations (>20 µmol/L).

CONCLUSIONS: The current reference ranges used in New Zealand for serum vitamin B₁₂ are set conservatively high. Serum MMA concentrations appear to allow better differentiation of a responsive condition than vitamin B₁₂ concentrations. Serum MMA concentrations <13 µmol/L indicate responsiveness to supplementation whilst concentrations <7 µmol/L indicate unresponsiveness. In the range 7–13 µmol/L, variation in response was observed and predictability of response is less certain, but supplementation is advisable.

CLINICAL RELEVANCE: The current reference ranges for vitamin B₁₂ responsiveness are conservatively high and lead to over-diagnosis of vitamin B₁₂ deficiency in ill-thriftiness of sheep.     

Congenital polycystic kidney disease in lambs

AIM: To describe the pathology and inheritance of a congenital polycystic kidney disease (PKD) of sheep.

METHODS: Mode of inheritance of PKD was investigated by evaluation of results of the disorder from planned matings in two consecutive years within subsets of a flock that had a high prevalence of PKD in lambs. Gross pathological and histopath- ological studies were based on tissues derived from this study. Haematoxylin and eosin (H&E)-stained paraffin sections of kidney, liver, extrahepatic biliary and pancreatic ducts, pancreas and epididymis were used to describe the lesions.

RESULTS: Twenty-five lambs affected by PKD, of both sexes, were born, numbers in accord with those expected for an autosomal recessive disorder in the population studied. In all cases for which tissues were available, the renal, bile ductal (intrahepatic and extrahepatic), pancreatic and epididymal tissues had widespread dysplastic changes and associated cyst formation.

CONCLUSIONS: The findings of renal cysts in conjunction with cysts in other organs are unifying features in many of the human and animal forms of PKD and suggest a related pathogenic and genetic base consistent with an autosomal recessive disorder.