Experimental 3-nitro-4-hydroxyphenylarsonic acid toxicosis in pigs

Five times the recommended dose of 3-nitro-4-hydroxyphenylarsonic acid (187.5 mg kg-1) induces a pathognomonic clinical syndrome in pigs. The main clinical features are not continually present but are inducible only by exercise. From day 11 on the experimental diet a nervous syndrome was inducible. This manifested as muscle tremors and clonic convulsive episodes. Paraparesis developed by day 22 and paraplegia by day 33. Liver arsenic levels plateaued at 5.4 +/- 1.3 mg kg-1. The experiment confirms field observations that 3-nitro produces a characteristic toxicological syndrome, which is distinct from that of arsanilic acid. It also confirms that 3-nitro has a higher absolute toxicity than arsanilic acid in pigs as well as a lower margin of safety.     

Neuropathology of experimental 3-nitro-4-hydroxyphenylarsonic acid toxicosis in pigs

Twenty pigs were fed a diet containing 187.5 mg kg-1 of 3-nitro-4-hydroxyphenylarsonic acid (3-nitro). Ten pigs were euthanized at intervals up to 29 days, 3-nitro was withdrawn from the diet of the remaining pigs on day 30, and these animals were subsequently euthanized at intervals up to 49 days after commencement of the experiment. A nervous syndrome characterized by clonic convulsive episodes inducible by exercise, developed at day 11. Paraparesis was apparent at day 22 progressing to paraplegia by day 33 (3 days after cessation of 3-nitro feeding). Histopathologic examination revealed myelin and axonal degeneration in the white matter of the spinal cord coincident with the onset of nervous signs. Marchi-positive degeneration was present in the dorsal funiculus at cervical level at day 22. Lesions intensified with increasing duration of toxicosis and while degenerate fibers were seen in all funiculi, there was preferential involvement of the fasciculi gracilis and cuneatus, the peripheral regions of the ventral and lateral funiculi, and a discrete area of the dorsal region of the lateral funiculus. Peripheral and optic neuropathies were evident from day 32 but were always mild and focal. The experiment establishes 3-nitro as a central-peripheral neurotoxicant of pigs.     

Roxarsone (3-nitro-4-hydroxyphenylarsonic acid) poisoning in pigs

Young pigs, six to ten weeks of age, from two unrelated swine operations were fed a grower ration obtained from a common commercial supplier. Following ingestion of the feed for approximately two weeks, pigs in both groups developed neurological disturbances characterized by blindness, ataxia, incoordination, muscle tremors, posterior paralysis, and quadriplegia. Vocalization described as “screaming” was also observed in several animals. Necropsy findings and tissue arsenic concentrations were consistent with a diagnosis of phenylarsonic acid poisoning. The liver and kidney contained an average arsenic content of 2.9 and 1.8 mg/kg (wet weight), respectively. The feed contained 38 mg of arsenic/kg corresponding to 133 mg roxarsone (3-nitro-4-hydroxyphenylarsonic acid)/kg. This level of roxarsone is approximately three to five times higher than the levels recommended for swine rations. The feed company had placed roxarsone in the ration at levels recommended for the less toxic arsanilic acid. It was assumed that the two organic arsenicals could be added to the rations interchangeably at the same level of formulation. The present investigation indicated that roxarsone is more toxic than arsanilic acid and the margin of safety in swine rations is low.     

Ulcerative cholecystitis produced by 3-nitro-4-hydroxy-phenylarsonic acid toxicosis in turkey poults

Naturally occurring ulcerative cholecystitis was present in a flock of 21-day-old turkey hens that were accidentally given at least 0.004% 3-nitro-4-hydroxy-phenylarsonic acid (3-NITRO) in their water for 2 days. Similar lesions were reproduced by administering 0.002% 3-NITRO to 2-day-old turkey poults for 6 days. Turkeys 31 days old were given up to 0.004% 3-NITRO in their water for 6 days, but no gall bladder ulcers were present in these poults. The toxicity to turkeys of 3-NITRO in the water appears to be age-dependent.     

Rates of change in liver copper concentration in cattle given a copper-deficient diet,with or without pre-treatment with tetrathiomolybdate,for evaluation of two parenteral copper supplements

AIM: To minimise the impact of initial variation in liver copper (Cu) on assessments of Cu supplements for cattle in depletion/repletion experiments.

METHODS: Efficacy of two Cu injections was assessed with 18 calves, weighing 200–250 kg, given a Cu-deficient barley diet, containing 4.1 mg Cu/kg dry matter (DM) and added molybdenum (3 mg/kg) and sulphur (3 g/kg). Initial liver biopsy Cu ranged from 3.15–14.17 mmol/kg DM and nine calves with the highest values were given three subcutaneous injections of 235 mg tetrathiomolybdate (TTM) after 42–46 days depletion to lower liver Cu. Untreated (L) and TTM-treated (H) calves were ranked separately for liver Cu after 50 days depletion and allocated to one of three treatments: 100 mg Cu given subcutaneously as CuCaEDTA in either a paraffin (CuP) or aqueous base (CuA) after 56 days depletion (Day 0) or no injection (O). Thereafter, plasma and liver biopsy Cu were measured every 2–4 weeks for 16 weeks. Responses in liver Cu to Cu injections were compared with and without log_e transformation and by linear regression.

RESULTS: Prior to Cu injection, the fractional decline in liver Cu concentration (FDLCu) after 50 days depletion was 0.64 (SE 0.066) and 0.80 (SE 0.090) in H and L calves, respectively (p=0.09) and mean liver Cu did not differ on Day ?6 (6.65 (SE 0.516) and 4.91 (SE 0.681) mmol/kg DM, respectively). Mean plasma Cu was higher in H than L calves on Day 0 (16.6 (SE 0.52) and 13.3 (SE 0.49) μmol/L, respectively (p<0.001)). Rates of decline in log_e liver Cu between Days 0–84 in treatments L and H were: 0.0138 and 0.0071 for Groups O; 0.0033 and 0.0016 for Groups CuP; 0.0073 and 0.0049 for Groups CuA (pooled SE 0.0014) mmol/kg DM/day, respectively. Between Days 84–114, FDLCu was uniformly high across experiments and groups (0.59 (SE 0.042)). Cu injections did not affect plasma Cu, which remained 3.1 (SE 0.41) umol/L higher in Experiment H than in L (p=0.017).

CONCLUSIONS: The use of rates of change in liver copper concentrations improved the assessment of efficacy for two parental copper supplements and that of pre-treatment with tetrathiomolybdate, which, contrary to expectation, slowed Cu turnover by mechanisms that remain unclear.     

Dynamics of uric acid concentration in the liver of cockerels subjected to model stress situations induced by fasting]

A model stress situation was created by nutrition deprivation. The experiment was performed with 130 cockerels of the initial breed White Leghorn. The experimental cockerels of the age of 18 to 24 days fasted for 24, 48 and 72 hours. Uric acid was determined in the supernatant of liver homogenate by means of a colorimetrically reductometric method with phospho-wolframic agent. A significant increase in the uric acid concentration in the liver of cockerels was found out during the model stress: the concentration of uric acid increased progressively from the initial 70.25 +/- 8.94 mg per 100 g to 93.15 +/- 9.66 mg per 100 g after 24-hour fasting, to 90.23 +/- 7.84 mg per 100 g after 48-hour fasting and even to 137.04 +/- 15.59 mg per 100 g after three-day fasting of 72 hours. After re-alimentation, the concentrations of uric acid were reduced to physiological values already within 24 hours.     

高铜对肉鸡肝线粒体膜通透性、脂类代谢及肝和肌肉铜含量的影响

选用1日龄健康AA肉鸡120只随机均分成4组,使用硫酸铜作为铜源,饲喂玉米-豆粕型基础日粮,对照组饲料铜含量为11mg/kg,3个试验高铜组饲料铜含量分别为110,220,330mg/kg,试验至60日龄结束,来探讨高铜日粮对肉鸡肝线粒体膜通透性、脂类代谢及肝和肌肉铜含量的影响。结果显示:(1)随着铜浓度的增加和/或饲养日龄的延长,线粒体膜通透性转换孔(Mitochondrial permeability transition pore,MPTP)逐渐扩大(P0.05);(2)血清总胆固醇(T-Ch)含量110,220mg/kg铜添加组低于对照组(P0.05),330mg/kg铜添加组极显著低于对照组(P0.01),而甘油三酯(TG)的含量在各高铜组均极显著低于对照组(P0.01);(3)与对照组相比,肝脏铜含量110,220mg/kg铜添加组有所增加(P0.05),330mg/kg铜添加组极显著增加(P0.01),肌肉中铜含量在各高铜组均极显著高于对照组(P0.01)。这说明高铜可造成肝线粒体不同程度的肿胀和损伤,改变肉鸡脂类的新陈代谢,明显增加肝和肌肉组织的铜含量。     

Changes in liver copper concentration of thoroughbred foals from birth to 160 days of age and the effect of prenatal copper supplementation of their dams

OBJECTIVES: To monitor the change in liver copper concentration of Thoroughbred foals from birth to 160 days of age and to determine the effects of supplementation by two injections of copper edetate given to dams in late gestation on the liver copper concentration of their foals at birth. PROCEDURE: Ten mares pregnant to the same stallion were randomised into two groups on the basis of age, liver copper concentration and expected foaling date. The treatment group mares were given 100 mg and 250 mg copper edetate intramuscularly during the ninth and tenth months of gestation respectively. Foals had liver biopsies taken weekly in the first month of life, then monthly for four months. Foals were euthanased at 160 days of age; liver samples were taken and the copper concentrations were determined. RESULTS: Two distinct patterns of age dependent decline in liver copper concentration were evident. The mean (+/- SD) liver copper concentration of the foals was high at birth (374 +/- 130 mg/kg DM), and for seven it declined to adult values by 160 days of age (21 +/- 6 mg/kg DM). In three foals the decline was at a slower rate than in the other seven and at 160 days of age the mean concentration was 162 +/- 32 mg/kg DM. Repeated measures analysis showed significant differences between each biopsy (P < 0.01) and between 'normal' and 'accumulator' foals (P < 0.002). Copper injections given to mares in late pregnancy had no effect on the liver copper concentration of foals at birth. CONCLUSIONS: The significance of the two patterns of age dependant decline in liver copper concentration is unknown. Parenteral copper supplementation of the dam in late gestation had no effect on the liver copper concentration of the foal at birth.     

硼对铜中毒肉用乌骨鸡肝铜含量及肝功能的影响

选用560只1日龄肉用乌骨鸡,采用二因子多水平随机化试验设计来探讨日粮硼对铜中毒肉鸡肝铜及肝功能的影响.试验中分别在玉米-豆粕型基础日粮中(50.9 mg/kg Cu,10.1 mg/kg B)添加549.1,749.1 mg/kg的铜和49.1,109.1 mg/kg的硼.结果表明,硼可在一定程度上抵御高铜对肝细胞的毒害,并减少肉鸡肝铜的蓄积量;说明硼对肉用乌骨鸡铜中毒具保护效应.     

Reduction of Salmonella typhimurium concentration in broiler chickens by milk or whey

Whey (5%) in the feed of chicks for the first 10 days of life reduced the mean log10 number of viable S. typhimurium from 5.68 in control chickens to 3.38 in whey-fed chickens. Lactose in drinking water or reconstituted dry milk (5% wt: vol) in drinking water reduced the mean log10 number of S. typhimurium to 2.60 and 2.11, respectively. Milk (5% wt: wt) in feed was not effective in reducing S. typhimurium colonization. The lack of effect of milk in the feed is believed to be because not enough lactose was provided at the 5% (wt: wt) concentration. Lactose in whey or nonfat dried milk offers alternatives to the use of pure lactose in preventing or lowering S. typhimurium numbers in young broiler chickens.     

Effect of infectious bursal disease virus insult on iron, copper, and zinc concentration in liver, bursa of Fabricius, spleen, pancreas, and serum of chickens

The effect of a systemic disease on the dynamics of iron, zinc, and copper in chickens fed ad libitum was examined by infecting 10-day-old specific pathogen-free chickens with infectious bursal disease virus (IBDV). Liver, bursa of Fabricius, pancreas, spleen, and serum were sampled in 10 controls and 10 challenged chickens at 3-day intervals postinfection (PI) for 15 days. The samples were analyzed using atomic absorption spectroscopy. Serum levels were similar to that reported in the literature. Concentrations of iron and zinc did not change significantly in the pancreas, but there was an increase in copper in infected pancreatic tissue on days 9 and 15 PI. Iron concentration in the spleen showed a significant increase on days 6, 9, and 15 PI, whereas zinc was only significantly increased on day 15 PI. There was no significant change in copper concentrations in the spleens of infected chickens vs. controls. This finding is in line with previously reported data. The results showed that the liver was not a major tissue where iron and zinc were sequestered, as previous data have shown in mammals. Instead, the bursa of Fabricius had significantly increased levels of both iron and zinc in infected tissue vs. control tissue from 9 days PI on. Furthermore, the bursa had increased levels of copper in the latter portion of the study. These findings suggest that the bursa of Fabricius rather than the liver is the major organ for metallic ion sequestering during IBDV infection.     

Effects of dried whey and copper sulfate on the growth responses to organic acid in diets for weanling pigs

Five 21-d to 28-d experiments involving 484 pigs weaned at 28 +/- 2 d of age were conducted to evaluate the effects of addition of organic acid to a fortified, corn-soybean meal diet (CS) or to a similar diet containing 15% dried whey (CSW) on performance of pigs. The effects of an antibiotic-sulfonamide combination (110 mg chlortetracycline, 110 mg sulfamethazine, 55 mg penicillin/kg) and the interactive effects of Cu sulfate (250 ppm Cu) and acid also were evaluated. The acid was a commercial product consisting of 96% organic acid (citric acid and Na citrate, 2:1). Treatments in Exp. 1 and 2 were factorial arrangements of the CS or CSW basal diets supplemented with 0 or 1% (Exp. 1) and 0, .5 or 1% (Exp. 2) of the acid product. Pigs fed diets containing whey consumed more feed (P less than .01) and gained weight faster (P less than .05), but they had feed/gain responses similar to those of pigs fed the CS diet. Addition of 1% acid improved (P less than .01) growth rate of pigs fed the CS diet but did not improve (P greater than .25) growth rate of pigs fed the CSW diet. Feed/gain was improved (P less than .01) by acid addition to both the CS and the CSW diets. Improvements in gain and feed/gain were similar for the two levels of acid. In Exp. 3 and 4, factorial combinations of 0 and 1% acid and 0 and 250 ppm Cu were evaluated in diets containing an antibiotic-sulfonamide combination. In addition, a negative control diet (no antibiotics, acid or Cu) was included. Pigs fed diets containing antibiotics gained faster and more efficiently (P less than .01) than those fed the control diet.(ABSTRACT TRUNCATED AT 250 WORDS)     

Distribution of iron and copper in the liver and spleen of veal calves in relation to the concentration of iron in the diet

The distribution of iron and copper in the liver and spleen of veal calves was determined in a group of calves fattened with a commercial milk replacer supplemented with 5 ppm iron and in a control group fed the unsupplemented milk replacer. No marked variations were detected in the distribution of iron and copper. A liver biopsy is therefore likely to be a representative sample of the whole liver with respect to iron and copper concentration.     

Effects of lipid sources,lysophospholipids and organic acids in maize-based broiler diets on nutrient balance,liver concentration of fat-soluble vitamins,jejunal microbiota and performance

1. Three experiments with a 2 × 2 × 2 factorial arrangement were conducted to evaluate maize-based diets for broilers containing different lipid sources [soybean oil (S) or beef tallow (T)] supplemented with or without lysophospholipids and organic acids on nutrient balance (Experiment I, evaluation period of 10–14 d), on liver concentration of fat-soluble vitamins, on jejunal microbiota (Experiment II, sampling at d 14) and on performance (Experiment III, accumulated periods of 1–14, 1–21 and 1–42 d).

2. A total of 1344 male chicks were used. In each experiment, the birds were allotted in a completely randomised design with 8 replications. The lysophospholipids were mainly composed of lysolecithins and the organic acids blend was constituted by lactic (40%), acetic (7%) and butyric acids (1%).

3. An interaction between lipid sources and lysophospholipids was observed on faecal apparent digestibility of lipid (ADL), which improved with lysophospholipids addition in T diets. Broilers fed on S had higher ADL and faecal apparent digestibility of nitrogen-corrected gross energy (ADGE_N).

4. It was not possible to demonstrate a significant treatment effect on the liver concentration of vitamins A and E, even with the differences in fatty acid profile between S and T.

5. Enterobacteria values were below the detection threshold. Lysophospholipid supplementation reduced gram-positive cocci in T-fed birds. S diets promoted lower total anaerobe counts compared with T diets, independent of additives.

6. S diets increased BW gain and feed:gain ratio in all evaluation periods. Lysophospholipids and organic acids improved feed:gain ratio at 1–21 d in T diets. Furthermore, main effects were observed for lysophospholipids and organic acids at 1–42 d, which increased BW gain and improved feed:gain ratio, respectively.

7. No positive interactions between additives were found.

     

高效液相色谱法检测奶牛血清中脂肪酸浓度

为了建立奶牛血清中脂肪酸检测方法,采用高效液相色谱法,异丙醇-正庚烷-2 mol/L磷酸(80∶20∶2)提取,正庚烷萃取,2-溴苯乙酮衍生化,流动相为乙腈-甲醇-水梯度洗脱,流速0.45 mL/min,色谱柱为Inert Sustain C18分析柱(150 mm×2.1 mm,3μm),紫外检测波长为244 nm。结果显示:二十碳五烯酸(C20∶5,EPA)、豆蔻酸(C14∶0)、二十二碳六烯酸(C22∶6, DHA)、花生四烯酸(C20∶4,AA)、亚油酸(C18∶2)、棕榈酸(C16∶0)、油酸(C18∶1)、正-十七烷酸(C17∶0)、硬脂酸(C18∶0)可以完全分离,峰形良好,精密度RSD≤2.96%,回收率在96%～99%之间。本方法稳定、可靠,适用于奶牛血清中脂肪酸浓度的检测。