Reduction of liver copper concentration by the organic arsenical, 3-nitro-4-hydroxyphenylarsonic acid

The interaction between 3-nitro-4-hydroxyphenylarsonic acid (roxarsone) and Cu was studied in a series of experiments with crossbred, broiler-type chicks. A fully fortified corn-soybean meal diet was fed in all assays. While roxarsone caused a marked reduction in liver Cu concentration, arsanilic acid (4-aminophenylarsonic acid), As2O3 and As2O5 were without effect. When structural analogs of roxarsone were studied, it was found that o-nitrophenol and 3-nitro-4-hydroxybenzoic acid also had no effect on liver Cu concentration in birds fed a high level of Cu. However, liver Co concentration was reduced by the addition of either o-nitrophenol or roxarsone to the diets of birds fed a high level of Co. It was concluded that arsenic per se had no effect on liver Cu accumulation or depletion, but that a chelate was probably formed between Cu or Co and the nitroso and hydroxyl groups of the ring portion of roxarsone. In addition to the reduction in liver Cu deposition, concentrations of Cu in the bile, brain, heart and pancreas of chicks were reduced by the addition of roxarsone to a high-Cu diet. Neither dietary nor intraperitoneally (ip) injected roxarsone had an effect on liver Cu concentration when Cu was injected ip. Therefore, both roxarsone and Cu had to be present in the diet for the liver Cu-lowering effect of roxarsone to be exerted. A further experiment was conducted with growing rats to determine the effect of roxarsone on Cu balance. Feeding roxarsone elevated Cu excretion in the urine but had no effect on Cu excretion in the feces.     

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.     

Fibrosing cholehepatitis in broiler chickens induced by bile duct ligations or inoculation of Clostridium perfringens.

The pathogenesis of fibrosing hepatitis causing condemnations in broiler chickens was investigated. Three to four week old broilers were inoculated via the hepatoenteric bile duct with saline washed suspensions of Clostridium perfringens (10(7) and 10(8) organisms). In another group of broilers, both bile ducts were ligated. The sequential development of liver and gall bladder lesions was studied at intervals ranging from 1-28 days postsurgery. The lesions were similar in both experiments in that the liver became mottled and swollen by five to seven days. Fibrinoid necrosis, heterophil and lymphocyte infiltration, bile duct hyperplasia and fibrosis with reticulin fiber proliferation occurred. By 14-17 days, the liver was enlarged, tan colored and firm with red and white foci. By 28 days, bile duct proliferation and fibrosis were massive with only a few hepatocytes remaining. The liver capsule was not involved. Jaundice was not present but the birds with ligated bile ducts excreted intensely yellow stained droppings after six to seven days. The gall bladder in inoculated birds was edematous and distended with flocculent or inspissated material. Clostridium perfringens was reisolated from gall bladder and/or liver of inoculated birds up to 28 days postsurgery. It is suggested that this organism plays a role in the pathogenesis of fibrosing cholehepatitis by inducing septic intrahepatic cholestasis.     

Experimental biliary cryptosporidiosis in broiler chickens

Biliary cryptosporidiosis was studied by inoculation of 2 x 10(5) Cryptosporidium baileyi oocysts (AU-B1 isolate) into the gall bladders of ten 6-day-old broiler chickens. Clinical signs of disease were not seen. Three of the 10 chickens developed biliary tract infections, based on histologic examination of tissue sections. Lesions seen in the gall bladders of these birds included epithelial hyperplasia and infiltration of the underlying connective tissue with mononuclear leukocytes. One of these birds also had involvement of the hepatic bile ducts. The bile ducts were mildly dilated and contained lesions similar to those seen in the gall bladder. Few to many cryptosporidia were present in the gall bladders and bile ducts of infected birds. Chickens may be of use in the study of biliary cryptosporidiosis, a common sequel to enteric infection in humans with human immunodeficiency virus infection.     

Investigations of toxicity episodes involving chemotherapeutic agents in Victorian poultry and pigeons

This series of case reports details observations on toxicity episodes in poultry due to a variety of chemotherapeutic agents. These problems arose owing to overdosage, variation in species susceptibility, potentiation of the toxic effects of one substance by the presence of another substance, and particular disease or other on-farm factors. Ignorance and accident were responsible for some of these situations. The episodes involved monensin, salinomycin, nicarbazin, sulphaquinoxaline, dinitolmide, dimetridazole, nitrofurans, streptomycin, and 3-nitro-4-hydroxyphenylarsonic acid.     

Emergence of fowl aviadenovirus C-4 in a backyard chicken flock in California

Fowl aviadenovirus (FAdV) species D and E are associated with inclusion body hepatitis (IBH); species C, serotype 4 (hereafter, FAdV4) is associated with hepatitis–hydropericardium syndrome (HHS) in young chickens. Outbreaks of HHS have led to significant losses in the poultry industry in several countries, predominantly in China. In April 2020, FAdV4 was detected in a remote backyard flock in California. In a mixed flock of chickens of various breeds and ages (6 mo to 2 y old), 7 of 30 were found dead within a week without premonitory signs. One additional bird died after the flock was relocated to fresh pasture, bringing the total mortality to 8 of 30 (27%). Postmortem examination of 3 birds revealed good body condition scores and active laying. One chicken had subtle hemorrhages throughout the liver, and the other 2 had diffusely dark mahogany livers. On histopathology, 2 chickens had hepatic necrosis with hepatocytes containing large, mostly basophilic, intranuclear inclusion bodies, identified by electron microscopy as 82.2-nm diameter adenoviral particles. Virus isolation and genomic sequencing performed on a liver sample revealed strains with 99.9% homology to FAdV4 isolates reported from China. To our knowledge, FAdV4 has not been reported in the United States to date. Furthermore, the chickens affected here were all adults and exhibited a variation of serotype 4 disease in which IBH was present but not hydropericardium.     

Arsenic-sulfur amino acid interactions in the chick

Experiments were conducted with growing crossbred chicks to determine the reasons why cysteine exacerbates roxarsone (3-nitro-4-hydroxyphenylarsonic acid) toxicity. A fortified corn-soybean meal diet that met or exceeded all nutrient requirements of the young chick was fed. While cysteine enhanced roxarsone toxicity, it had little effect on the toxicity of the inorganic arsenicals As2O3 and As2O5. The toxicity of another pentavalent organic arsenical, phenylarsonic acid, was also exacerbated by cysteine. In contrast, the growth-depression resulting from feeding the trivalent form of phenylarsonic acid, i.e., phenylarsine oxide, was not affected by dietary addition of cysteine. Supplementation of the diet with cystine, methionine or K2SO4 did not exacerbate roxarsone toxicity. Reduced glutathione (GSH), however, slightly increased the gain/feed depression resulting from feeding 300 mg roxarsone/kg diet. When injected ip 1) roxarsone and cysteine, or 2) roxarsone and ascorbic acid killed 100 or 60% of the birds, respectively, within 48 h postinjection. Few (6.7%) deaths resulted from ip injections of the same level of roxarsone alone. Therefore, the potentiation of toxicity requires pentavalent organic arsenicals and compounds that can act as reducing agents. We concluded that cysteine exacerbates roxarsone toxicity by reducing it to the more toxic trivalent state.     

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.     

Pancreatic necrosis and ventricular erosion in adenovirus-associated hydropericardium syndrome of broilers

Seven 19-day-old broiler chickens affected with hydropericardium syndrome (HPS) with pancreatic necrosis and gizzard erosions were investigated pathologically and virologically. Mortality increased after 13 days of age in a flock on a broiler farm. The mortality rate of the flock reached 10% by 19 days of age. Macroscopically, the chickens had hydropericardium (the characteristic gross change of HPS), pinpoint white foci in the pancreas, and ventricular erosions. Histologically, the chickens had multifocal hepatic necrosis with intranuclear inclusions in hepatocytes, a marked increase of macrophages in the spleen and lung, mild epicardial edema, multifocal necrosis of pancreatic acinar cells with intranuclear inclusions, focal necrosis of the ventricular koilin layer, and degeneration of the ventricular glandular epithelium with intranuclear inclusions. Immunohistochemically, intranuclear inclusions in the liver, pancreas, and ventriculi were stained positively against group I avian adenovirus (GIAAV) antigens. Ultrastructurally, 67-nm diameter viral particles were present in intranuclear inclusions. Virologically, serotype 4 of GIAAV was isolated from the liver, heart, and kidney of affected chickens. The pathologic changes of the present cases differ from previous cases of HPS; therefore, the present strain of GIAAV may have different pathogenicity for chickens than the previous virus strain of HPS.     

Effects of aflatoxin B1 and fumonisin B1 on body weight, antibody titres and histology of broiler chicks

1. Our objective was to evaluate the toxic effects of aflatoxin B1 (AFB1) and fumonisin B1 (FB1), administered singly or in combination to broilers. 2. Feeds were prepared with concentrations equal to 0, 50 and 200 microg AFB1/kg, and/or 0, 50 and 200 mg FB1/kg, and offered to broiler chicks from 8 to 41 d of age. The experimental design was totally randomised, in a 3 x 3 factorial arrangement with 9 treatments and 12 birds per treatment. Animals were vaccinated against Newcastle disease on d 14 of life and killed at 41 d. 3. Compared with controls, all mycotoxin-treated groups at 41 d had lower body weight and weight gain, and higher relative heart weight. The relative weight of the liver increased only in birds fed diets containing 200 mg FB1, singly or in combination with AFB1. 4. At 35 d, all groups receiving mycotoxin-treated rations had reduced geometrical mean antibody titres, with birds from groups fed combinations of AFB1 and FB1/kg having even lower values, when compared to the other groups. 5. Histological changes were observed only in liver from birds fed mycotoxin-contaminated rations, and in kidneys of birds fed the diet containing 200 microg AFB1 and 200 mg FB1/kg. Main alterations included vacuolar degeneration and cell proliferation of bile ducts in the liver, and hydropic degeneration in renal tubules in the kidneys. 6. We concluded that AFB1 and FB1 in combination have primarily additive effects on body weight, liver structure and immunological response of broilers at the concentrations used.     

Persistence of the V4 strain of Newcastle disease virus in an open-range flock of chickens

The V4 strain of Newcastle disease virus was introduced into a small open range flock of bantam chickens, by dosing half the birds directly into the crop. As indicated by rises in titres of haemagglutination inhibition antibody, the virus spread to the uninoculated birds and persisted in the flock for two years, infecting chickens that were introduced by natural brooding and rearing. All new clutches of chicks seroconverted by 80 days of age, and the titres of adult birds showed a concurrent rise, suggesting that the chicks were amplifying the virus. The modes of spread and of persistence of the virus were not determined; although cloacal swabs were taken regularly, only one yielded virus. Antibody titres of the inoculated birds remained above the presumptive protective level of 3 (log2) for over a year, whereas the titres of birds infected by contact were generally less than 3.     

An unusual case of inclusion body hepatitis in a cockerel

This report describes inclusion body hepatitis in a 12-week-old leghorn-type cockerel. The cockerel had come from a specific-pathogen-free flock and was reared in a positive-pressure isolator until 6 weeks old. Grossly, the cockerel was jaundiced, and the liver was swollen and had small white foci throughout. Histologically, there were foci of hepatic necrosis and basophilic inclusion bodies in the nuclei of numerous hepatocytes. A fowl adenovirus, isolated from the liver, was found to belong to serotype 2/12. The bursa of Fabricius had some degenerative changes, but these were not consistent with infectious bursal disease, nor was there evidence of seroconversion to infectious bursal disease virus in the remaining chickens.     

Evaluation of two mycotoxin binders to reduce toxicity of broiler diets containing ochratoxin A and T-2 toxin contaminated grain

In order to assess ochratoxin A (OA) and T-2 toxin (T-2) binding ability of two commercial sorbents, both in vitro and in vivo trials with broilers were performed. Crude OA and T-2 extracts from contaminated grain were used to assess in vitro binding ability of two sorbents (Zeotek [Zk] and Mycofix [Mx]), by quantifying free mycotoxin through an enzyme-linked immunosorbent assay (ELISA) test. For in vivo trial, a 3 x 2 x 2 factorial arrangement was used for this experiment, being the factors: adsorbents (none, Zk, and Mx), OA (0 and 567 parts per billion [ppb]) and T-2 (0 and 927 ppb). OA and T-2 contaminated wheat and corn, respectively, were added to sorghum-soybean meal diets to meet 567 ppb of OA and 927 ppb of T-2. Mycotoxins were fed alone or combined in treatments. After 21 days, blood chemistry, gross, and histological evaluations were performed. Relative weights of liver, kidney, and bursa of Fabricius were obtained. Zk had the highest OA and T-2 in vitro binding ability (100% and 8.67%, respectively). Chickens fed OA with or without sorbents had a lower body weight and feed intake reduction. However, those birds fed T-2 were partly protected by a sorbent. Birds fed both toxins showed toxic additive effects, and no protection of any adsorbent was observed. A significant reduction in plasma proteins, albumin, and globulins was a characteristic observed in all birds fed diets with OA both with or without adsorbents. Uric acid level in blood was increased in all chickens fed OA-contaminated diets. Histological findings observed in birds fed OA-contaminated diets were necrosis of kidney tubular cells, swollen and necrotic hepatocytes, bile ducts hyperplasia, and increased diameter of proventriculus glands. In birds that received T-2 alone, only the liver, with the same kind of lesions, was affected. According to these results, it can be concluded that there is not a relation between in vitro and in vivo trials. OA toxic effects could not be counteracted by any sorbent. T-2 toxicity could be partially counteracted by an adsorbent used in this research.     

Pathology of spontaneous colibacillosis in a broiler flock

Forty-eight of 134 chickens collected from a flock on a broiler farm were diagnosed pathologically and microbiologically to have colibacillosis. Both acute septicemia (seven birds, 1 to 36 days old) and subacute serositis (41 birds, 5 to 57 days old) were found. The former consisted of necrosis with fibrinous exudates in the ellipsoids and lymphoid follicles of the spleen, and fibrinous thrombi in sinusoids of the liver with occasional necrosis of hepatic cells. The latter had fibrinopurulent inflammation with granulomatous changes in the serosal tissues--including the epicardium, pericardium, and hepatic peritoneal sac--accompanied by septicemic lesions in the spleen and liver. Respiratory lesions (airsacculitis, pneumonia, and tracheitis) were noted in most chickens affected with acute septicemia and subacute serositis. Degenerative changes also were observed in the bursa of Fabricius.     

Feeding of Ferrets with the Raw Meat and Liver of Chickens Chronically Poisoned with Toxic Groundnut Meal

Chickens were fed a ration containing 30 per cent of toxic groundnut meal for up to six weeks. The concentration of aflatoxin (toxic metabolites of Aspergillus flavus) in the above ration was 3.06 p.p.m. At the end of 2nd, 4th or 6th week the birds were killed. The meat was removed from the bones and put through a meat grinder. The livers of three groups were pooled together. Three control groups of birds kept on commercial pellets were treated similarly.

Female ferrets, two years of age, were used in the present study. They were divided into four groups. The first three groups were given for one month meat from chickens fed the toxic ration for 2, 4, and 6 weeks, respectively. Each of these three groups contained one control ferret that was fed with the meat of chickens fed a commercial ration for a similar period of time. One half of the 4th group was fed pooled liver from intoxicated birds and one half was fed liver from control birds.

No significant changes in the ferret tissues were observed as a consequence of feeding them with the meat or liver from the chickens chronically poisoned with toxic groundnut meal.

     

IMPORTANCE OF ACUTE DEATH SYNDROME IN MORTALITIES IN BROILER CHICKEN FLOCKS

SUMMARY A survey of the weekly causes of mortality was carried out in a flock of 64,000 broiler chickens in Western Australia. Some 6.86% of the flock died or was culled in the period from day-old to slaughter and were examined post mortem. The most important cause of death, acute death syndrome (ADS) (36%), occurred during all weeks of production and 74% of affected birds were male. A comparison of liver biotin content made between 120 ADS-affected birds and 60 clinically normal flockmates established that the biotin status of ADS-affected birds was adequate.     

A TRANSMISSIBLE CHICKEN TUMOUR ASSOCIATED WITH RETICULOENDOTHELIOSIS VIRUS INFECTION

Histiocytic lymphosarcomas of the intestine, liver, spleen and sciatic nerve were found at necropsy in a 36-week-old laying hen that was culled from a flock of 1800 birds because of emaciation. Type C particles were observed in ultrathin sections of liver and spleen. The serum of the hen contained reticuloendotheliosis virus (REV) antigen, and antibody against REV, but lacked antibodies reactive with Marek's disease virus or subgroups A and B of Rous sarcoma virus. The tumour was transmitted to chickens using a suspension of the initial tumours. These experimental tumours were then transmitted to further chickens, using cultured spleen cells, viable spleen cells that had been stored frozen, and disrupted spleen cells. The tumours, which developed after incubation periods as short as 2 weeks, were histologically similar to those in the original hen. A few chickens also developed feather abnormalities. The chickens with experimentally transmitted tumours developed antibody against REV and REV antigen was demonstrated in cultured cells from these chickens. The chickens failed to develop antibody against Rous sarcoma virus and only 1 of 29 developed antibody against Marek's disease virus.     

An outbreak of fowl cholera in ring-necked pheasants (Phasianus colchicus)

A total of 120 ring-necked pheasants from a 3000-bird flock in Zeeland, MI, died over a 3-day period. Clinical signs included sudden death, diarrhea, and limping. At necropsy, hepatomegaly with multifocal cream-colored foci randomly distributed throughout the parenchyma was observed in diseased birds. Additionally, the spleen was enlarged up to three times its normal size and had a marbled appearance. Microscopically, there was multifocal splenic and hepatic necrosis with intralesional rod-shaped bacteria. Pasteurlla multocida serotype 3/4 was isolated from liver and spleen. In this paper, we report an outbreak of acute fowl cholera in ring-necked pheasants.     

Cholangiohepatitis in broiler chickens in Japan: histopathological, immunohistochemical and microbiological studies of spontaneous disease

Forty-five broiler carcasses from 6 different flocks were condemned due to liver lesions at processing meat inspection, and collected for pathological and bacterial examinations. All affected chickens showed liver enlargement with discolouration and an apparent acinar pattern. The enlarged gallbladder and the extrahepatic bile ducts contained yellow inspissated cream-coloured material. Histopathologically, extensive proliferation of bile ductules with fibrosis was observed in interlobular connective tissue, and it spread to form bridges with adjoining triads. Destruction and obstruction of portal bile ducts with multiple granulomas due to bacterial infection and outflow of the bile were frequently observed. Many Gram-positive bacilli were seen in the lesions, and they were identified as Clostridium perfringens by indirect immunofluorescence staining technique. Clostridium perfringens was isolated from affected livers. These findings are consistent with cholangiohepatitis. Therefore, it is suggested that C. perfringens might be important in the pathogenesis of cholangiohepatitis in broiler chickens.