Chemical fingerprinting of naphthenic acids and oil sands process waters-A review of analytical methods for environmental samples.

This article provides a review of the routine methods currently utilized for total naphthenic acid analyses. There is a growing need to develop chemical methods that can selectively distinguish compounds found within industrially derived oil sands process affected waters (OSPW) from those derived from the natural weathering of oil sands deposits. Attention is thus given to the characterization of other OSPW components such as oil sands polar organic compounds, PAHs, and heavy metals along with characterization of chemical additives such as polyacrylamide polymers and trace levels of boron species. Environmental samples discussed cover the following matrices: OSPW containments, on-lease interceptor well systems, on- and off-lease groundwater, and river and lake surface waters. There are diverse ranges of methods available for analyses of total naphthenic acids. However, there is a need for inter-laboratory studies to compare their accuracy and precision for routine analyses. Recent advances in high- and medium-resolution mass spectrometry, concomitant with comprehensive mass spectrometry techniques following multi-dimensional chromatography or ion-mobility separations, have allowed for the speciation of monocarboxylic naphthenic acids along with a wide range of other species including humics. The distributions of oil sands polar organic compounds, particularly the sulphur containing species (i.e., OxS and OxS2) may allow for distinguishing sources of OSPW. The ratios of oxygen- (i.e., Ox) and nitrogen-containing species (i.e., NOx, and N2Ox) are useful for differentiating organic components derived from OSPW from natural components found within receiving waters. Synchronous fluorescence spectroscopy also provides a powerful screening technique capable of quickly detecting the presence of aromatic organic acids contained within oil sands naphthenic acid mixtures. Synchronous fluorescence spectroscopy provides diagnostic profiles for OSPW and potentially impacted groundwater that can be compared against reference groundwater and surface water samples. Novel applications of X-ray absorption near edge spectroscopy (XANES) are emerging for speciation of sulphur-containing species (both organic and inorganic components) as well as industrially derived boron-containing species. There is strong potential for an environmental forensics application of XANES for chemical fingerprinting of weathered sulphur-containing species and industrial additives in OSPW.

Experimental reproduction of Mycoplasma gallisepticum disease in chukar partridges (Alectoris graeca).

An outbreak of conjunctivitis and severe respiratory disease occurred in an integrated chukar partridge (Alectoris graeca) operation that involved about 8000 birds. The main clinical features were conjunctivitis and sinusitis and frequent mouth breathing, but almost no gasping or coughing. In 1000 breeders, egg production declined from 73% to 20%. Morbidity reached 100%, and losses from mortality and culling approached 60%. At necropsy, a conjunctivitis (often bilateral) and extensive caseated sinusitis were common. There was an occasional slight mucoid tracheitis, but no significant air sac lesions were noted. Mycoplasma gallisepticum, designated strain GM1125, was isolated and identified. Exposure of susceptible chukars to GM1125 reproduced the field disease. GM1125 was reisolated from the conjunctiva of all exposed birds 12 days postinfection, but infrequently from there or the respiratory system 36 days postexposure, even though clinical disease was still present. The experimental disease was confined to the conjunctiva and the upper respiratory tract. An occasional mucoid tracheitis was noted, but generally, the lungs and air sacs were not involved. Infection was followed by an appreciable serological response to M. gallisepticum.

Neoplasms and related proliferative lesions in control Sprague-Dawley rats from carcinogenicity studies. Historical data and diagnostic considerations.

Historical data are presented for neoplasms and related proliferative lesions from 1,170 Sprague-Dawley rats that served as controls in 9 carcinogenicity (2 year) studies conducted in the Safety Evaluation Facility of Ciba-Geigy Corporation, Summit, New Jersey. The most common neoplasm was pituitary adenoma, which occurred in 62.2% of the male and 84.7% of the female rats. Incidences of other neoplasms that occurred in more than 6.0% of the rats were, for males, benign pheochromocytoma (19.0%), cutaneous keratoacanthoma (7.9%), pancreatic islet cell adenoma (7.5%), benign testicular interstitial cell tumor (6.5%), and thyroid C-cell adenoma (6.5%). For females these incidences were mammary fibroadenoma (31.3%), mammary adenocarcinoma (16.8%), and mammary adenoma (6.5%). Focal cortical hypertrophy/cystic degeneration of the adrenal, a focal nonneoplastic lesion of zona fasciculata cells that often degenerate into large cysts, was present in 23.4% of all male and 82.7% of all female rats. Criteria for the differential diagnoses of selected neoplasms and related lesions are presented.

Delineation of the lateral spread of Mycoplasma gallisepticum infection in chickens.

Lateral spread of S6 strain Mycoplasma gallisepticum (MG) was studied in small populations of chickens. One experimentally exposed bird served as the source of infection, and the presence of MG-agglutinating antibody was evidence of infection in individuals. The results were subjected to survival data analysis. In the seven experiments, four similar but not identical phases of lateral spread were observed: phase 1, a generally long latent phase (median 15, range 12-21 days) before antibody was first detected in the MG-inoculated bird; phase 2, a generally short period (median 1, range 1-21 days) in which infection gradually appeared in 5-10% of the population; phase 3, a fairly constant characteristic phase (median 24, range 7-32 days) in which 90-95% of the remaining population developed MG antibody; phase 4, a generally short terminal phase (median 4, range 3-19 days) in which the remainder of the population became positive. Increasing the population density increased the rate at which lateral spread occurred.

Toxicity of chloroform and bromodichloromethane when administered over a lifetime in rats.

Chloroform (CHCl3) and bromodichloromethane (CHBrCl2) are the two most common haloorganic contaminants of chlorinated drinking water. A significantly increased incidence of hepatic neoplastic nodules was found in female rats when either of these compounds was administered in drinking water to Wistar rats throughout their life-span. Hepatic adenofibrosis was also produced by chronic ingestion of these two halomethanes.

The effects of Love Canal soil extracts on maternal health and fetal development in rats.

The effects of a solvent extract of the surface soil of the Love Canal chemical dump site, Niagara Falls, New York, and of a natural extract, or leachate, which is drained from the canal for treatment, on the maternal health and fetal development were determined in rats. The solvent extract, which was contaminated with 2,3,7,8-tetrachlorodibenzo-p-dioxin (2, 3,7,8-TCDD) at 170 ppb and numerous other chlorinated organic compounds with the primary identified components being the isomers of benzenehexachloride (BHC), was dissolved in corn oil and administered by gavage to pregnant rats at 0,25,75, or 150 mg crude extract/kg/day on Days 6-15 of gestation. A 67% mortality was observed at the highest dose. The rats were sacrificed on Day 20. Dose-related increases in relative liver weight accompanied by hepatocyte hypertrophy were observed at all dose levels. Fetal birthweight was decreased at 75 and 150 mg extract/kg/day. No major treatment-related soft tissue or skeletal malformations, except for delayed ossification, were observed. Based on literature values for BHC, all of the observed toxicity could be accounted for by the BHC contaminants of the extract. The crude organic phase of the leachate was administered to pregnant rats at 0,10,100, or 250 mg/kg/day as described above. Maternal weight gain decreased at 100 and 250 mg/kg/day, accompanied by 5 and 14% maternal mortality, and 1 and 3 dead fetuses, respectively. Early resorptions and the percentage of dead implants increased whereas fetal birthweights were decreased at 250 mg/kg/day. No major treatment-related soft tissue or skeletal malformations, except for delayed ossification, were observed. The primary components of the complex leachate by mass were tetrachloroethanes; however, 2,3,7,8-TCDD, which was present at 3 ppm, probably accounted for all the observed toxicity.

Prevalence of Mycoplasma gallisepticum and M. synoviae in commercial layers in southern and central California.

The prevalence of Mycoplasma gallisepticum (MG) and M. synoviae (MS) in commercial pullet and layer flocks in Southern and Central California was estimated by testing serum and egg-yolk samples from 360 sample flocks in Southern California and 41 sample flocks in Central California. Data relating to potential risk factors associated with MG and MS infections were collected. The estimated true prevalence rate of MG was 73% in Southern California and 3% in Central California. The estimated true prevalence rate of MS was 91% in Southern California and 32% in Central California. Compared with uninfected flocks, MG-infected flocks in Southern California were significantly older and were medicated less (P less than 0.05). More managements were under a multiple-age system, more flocks had molted, more were vaccinated with F-strain, and more had concurrent infection with MS (P less than 0.05). Only one sample flock in Central California was MG-infected; none were vaccinated with F-strain. In Southern California, MS-infected flocks were older than uninfected flocks, more had molted, more were medicated, and more had concurrent infection with MG (P less than 0.05). In Central California, MS-infected flocks did not differ significantly from uninfected flocks in any factor examined; the lack of statistical significance may be due to small sample size.

Subchronic oral toxicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin in the guinea pig: comparisons with a PCB-containing transformer fluid pyrolysate.

In contrast to the well-characterized acute toxicity of the environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TCDD) in the guinea pig, the effects of prolonged po exposure in this species are unknown. The present report describes the results of administration to guinea pigs of 2,3,7,8-TCDD in the feed at levels of 0, 2, 10, 76, or 430 ppt for up to 90 days. Additional aims were to examine recovery following prolonged 2,3,7,8-TCDD exposure in the guinea pig and to generate data to facilitate comparison of the previously reported toxicity of a transformer fluid pyrolysate with that of pure 2,3,7,8-TCDD. Animals receiving 430 ppt 2,3,7,8-TCDD exhibited body weight loss, thymic atrophy, liver enlargement, and 60% mortality by Day 46 (males) and by Day 60 (females), when surviving animals in this group were sacrificed. Total 2,3,7,8-TCDD consumption was approximately 1.3 and 1.9 micrograms/kg, respectively. Animals receiving 76 ppt 2,3,7,8-TCDD for 90 days (total 0.44 microgram/kg) exhibited a decreased rate of body weight gain and increased relative (to body) liver weights. Male animals also displayed a reduction in relative thymus weights and elevated serum triglycerides, while females exhibited hepatocellular cytoplasmic inclusion bodies and lowered serum alanine aminotransferase activities. Toxic effects were generally similar to those observed after acute 2,3,7,8-TCDD administration. No dose-related alterations were seen in animals receiving either 10 ppt (total 0.06 micrograms/kg) or 2 ppt (total 0.01 micrograms/kg) for 90 days, establishing a no-observed-effect level of approximately 0.65 ng 2,3,7,8-TCDD/kg/day. In the recovery study, groups of guinea pigs were administered 430 ppt 2,3,7,8-TCDD for 11, 21, or 35 days and then allowed to recover for an additional 79, 69, or 55 days, respectively. Treatment-related mortality in each group was 0, 10, and 70%, respectively, by Day 90. An effective LD50 of 0.8 microgram 2,3,7,8-TCDD/kg for prolonged exposure was calculated on the basis of these results, a value lower than those previously reported from this laboratory for acute exposure. The results also suggested a possible lowering of the body weight "set point" following 2,3,7,8-TCDD exposure. Comparison of the present findings with those previously reported for a transformer fluid pyrolysate containing a mixture of polychlorinated aromatic species indicated both a greater variety of toxic effects and flatter dose-response relationships for the pyrolysate in the guinea pig.

Observations on commercial layers vaccinated with Mycoplasma gallisepticum (MG) bacterin on a multiple-age site endemically infected with MG.

A commercially available inactivated Mycoplasma gallisepticum (MG) bacterin was administered to chickens on a multiple-age farm endemically infected with MG. A total of 3400 MG-free pullets were vaccinated with the MG bacterin at 19 weeks of age, and 4300 unvaccinated pullets served as controls. The vaccinated group became serologically positive by the rapid plate agglutination (RPA) test within 3 weeks, and the unvaccinated group became positive in 7 weeks. The hemagglutination-inhibition test responses were observed at approximately the same time as the RPA in both of the groups. Egg production and mortality through 50 weeks of age did not differ significantly between the two groups. MG was isolated from birds of the vaccinated and control groups near the termination of the study.

Histologic characterization of the involuting bursa of Fabricius in single-comb white Leghorn chickens.

Bursas from specific-pathogen-free white leghorn chickens of both sexes were examined at several intervals from hatching to 28 weeks of age. No histologic alterations other than scattered atrophic or cystic follicles were observed through 20 weeks. Obvious involution, first noted at 24 weeks, was in early stages in females and quite advanced in males. Involution was essentially complete by 26 weeks, and only cicatrized vestiges of bursas were present at 28 weeks of age. Gross manifestations included bursal atrophy, variable yellowish discoloration of the mucosa, and matting or total loss of identity of the mucosal plicae. Histologic characteristics of involution are summarized by the following approximate sequence: atrophy and exfoliation of plica epithelium; subepithelial stromal fibrosis; fusion and ultimate collapse of plicae; liquefactive necrosis of first medullary then cortical elements of follicles, which seemed to progress from basal to apical portions of the plicae; progressive proliferation of stromal connective tissue and infiltration of macrophages into areas occupied by necrotic follicles; and, finally, complete fibrous organization of luminal debris, leaving a firm nodule formed by a contracted muscularis surrounding the cicatrized remains of the mucosa.

Lifetime toxicity of chloroform and bromodichloromethane when administered over a lifetime in rats.

Chloroform (CHCl3) and bromodichloromethane (CHBrCl2) are the two most common haloorganic contaminants of chlorinated drinking water. A significantly increased incidence of hepatic neoplastic nodules was found in female rats when each of these compounds was administered in drinking water to Wistar rats throughout their life span. Hepatic adenofibrosis was also produced by chronic ingestion of these two halomethanes.

Comparative susceptibility of embryonated hen eggs and pig lung alveolar macrophages to chlamydial infection.

Pig lung alveolar macrophages provided a suitable cell culture system for primary isolation of ovine chlamydia and sustained their growth through several passages. In centrifuged preparations, titres of established strains compared favourably with those obtained in fertile eggs, and in parallel titrations on primary isolation, evidence of chlamydial infection was detectable in macrophages several days earlier than in eggs.

Esterase A is a proteinase from rat urine that can activate plasminogen.

A proteinase which can activate human, dog and rat plasminogen to plasmin has been isolated from the urine of female rats, using affinity chromatography on benzamidine-coupled Sepharose. Inhibition by diisopropylfluorophosphate, tosyl-L-lysine chloromethylketone and benzamidine classified the enzyme as trypsin-like. The proteinase has weak activity on alpha-casein and hemoglobin, but will not lyse fibrin clots. It readily cleaves arginyl amides, including synthetic substrates specific for human glandular kallikrein and other serine proteinases. A chromogenic substrate for human urokinase (pyro Glu-Gly-Arg-pNA) is a poor substrate for the rat proteinase. Characteristics of the enzyme, such as its molecular weight (25 900), kinetic parameters and inhibition by aprotinin, indicate that this proteinase is esterase A, described by several investigators. Esterase A is shown not to be a true urinary plasminogen activator but rather is a unique arginine-specific proteinase. Urokinase-like and kallikrein-like activity are part of a broader proteolytic activity displayed by this enzyme.

Subchronic exposure of mice to Love Canal soil contaminants.

The health hazard potential of soil collected from the surface of the Love Canal chemical dump site in Niagara Falls, New York, was assessed in 90-day exposure studies. Female CD-1 mice were exposed to two concentrations of the volatile components of 1 kg of soil with and without direct soil contact. Control mice were identically housed but without soil. The soil was replaced weekly and 87 compounds were detected in the air in the cages above fresh and 7-day-old soil as analyzed by gas chromatography/mass spectrometry. The concentration of many of these compounds decreased during the 7-day exposure cycle. Histopathologic, hematologic, and serum enzyme studies followed necropsy of all mice. There was no mortality of mice exposed for up to 90 days under any condition. Thymus and spleen weights relative to body weight were increased after 4 weeks of exposure by inhalation but not after 8 or 12 weeks of exposure. alpha-, beta-, and delta- Benzenehexachlorides , pentachlorobenzene, and hexachlorobenzene were detected in liver tissue from these animals. Mice exposed to 5- to 10-fold elevated concentration of volatiles had increased body and relative kidney weights. There was no chemically induced lesion in any animal exposed only to the volatile soil contaminants. Mice exposed by direct contact with the soil without elevated volatile exposure had increased body (10%) and relative liver weights (169%). Centrolobular hepatocyte hypertrophy, which involved 40 to 70% of the lobules, was observed in all mice in this group.(ABSTRACT TRUNCATED AT 250 WORDS)

Growth of chlamydia in pig lung alveolar macrophages; preparation of macrophages and demonstration of growth.

The harvesting, storing and utilisation in tissue culture of lung alveolar macrophages from 2-3 weeks old piglets were investigated. An average yield of 4 X 10(8) cells was obtained, which in culture usually remained healthy for at least 8 days. In liquid nitrogen at -173 degrees C about 50% of the cells were still viable after 3 months. Pig cells sustained the growth of ovine Chlamydia psittaci and a rising titre was demonstrated over 4 passages.

Potentiation of the toxic effects of acetaminophen in mice by concurrent infection with influenza B virus: a possible mechanism for human Reye's syndrome?

Using weanling mice of two different genetic strains we demonstrated a potentiation of the toxic effects of acetaminophen by prior infection with influenza B virus. The C57BL/6N (B6) strain of mice is genetically predisposed to increased toxicity from acetaminophen when the hepatic cytochrome P-450 mixed function oxidase system is preinduced. When B6 animals are pretreated with influenza B virus and an mixed function oxidase system inducing agent before administering acetaminophen, we observed a significant incidence of atypical "fatty" liver pathology on light microscopy similar to the microvesicular steatosis seen in human Reye's syndrome. Electron microscopic changes in the liver of these animals resemble those published to date in human Reye's syndrome.

Experimental chlamydial pneumonia in pigs.

Naturally occurring disease in pigs associated with chlamydial infections has not been reported in Britain, though evidence of chlamydial challenge has been demonstrated in two separate serological surveys. An isolate of Chlamydia psittaci (28/68) from an ovine pneumonia produced pneumonia in pigs following intratracheal inoculation. Transient pyrexia at 24 hr was followed by increased respiratory rates and inappetance which lasted for a further 48 hr in challenged pigs. Histologically acute exudative reactions were present in the lungs by 24 hr with proliferative changes predominating after 10 days. While variations in the concentrations of inocula were reflected by corresponding increases and/or decreases in gross lung damage, clinical signs and histological reactions were unaltered. Chlamydial organisms were recovered only from lung tissues.

Trifluorinated ether anesthetic lethality in rats: the role of bacterial infection.

The lethal effects of the fluorinated ether anesthetics fluroxene (2,2,2-trifluoroethyl vinyl ether) and its ethyl (TFEE) and allyl analogues in male Wistar rats have previously been demonstrated to be potentiated by specific hepatic microsomal cytochromes P-450, and mediated by the common metabolite 2,2,2-trifluoroethanol (TFE). We report here that administration of lethal combinations of anesthetic and cytochrome P-450-inducing agents or of lethal doses of TFE (0.21 g/kg and higher) to rats caused decreased white blood cell counts, necrosis of sternum bone marrow cells and lymphocytes in the thymic cortex, and resulted in Escherichia coli contamination of the blood, lungs, liver, and kidneys of treated rats. Control animals in identical environments were free of bacterial contamination. Pretreatment of rats with the antibiotic tetracycline-HCl in the drinking water (0.6 g/liter) from 24 hr before anesthetic or TFE administration significantly diminished the mortality. With TFEE and beta-naphthoflavone induction, mortality was reduced from 85 to 30% by the antibiotic. However, the antibody plaque assay following immunization with sheep erythrocytes indicated that the primary humoral immune response to a thymus-dependent antigen was not impaired in treated rats. These results considered together indicate that metabolic formation of TFE from the anesthetic agents produced a decreased host resistance with subsequent increased susceptibility to bacterial infection. If not administered the antibiotic, the animals succumbed to the infection.