BACKGROUND: Free cyanide is a potent toxic agent in the aquatic environment.  Freshwater fish are the most cyanide-sensitive group with high mortality at free cyanide concentrations above 20 μg/L. Exposure to cyanide ions can cause stress, increased mortality and place an appreciable metabolic load on fishes. Rhodanese is a ubiquitous mitochondrial enzyme in both prokaryotes and eukaryotes that detoxifies cyanide (CN-) by converting it to thiocyanate (SCN). OBJECTIVES: The purpose of this investigation was to determine and compare the pattern of tissue distribution of Rhodanese in different tissues of four native Barbus fish including Mesopotamichthys sharpey, Tor grypus, Luciobarbus xanthopterus and Luciobarbus barbulus. METHODS: Fishes (10 from each species) with length of 32.5 ± 6.5 and weight of 440 ± 110   were collected from five major fishing reservoirs of Karoun River including Gotvand, Shushtar, Molasani, Darkhoine and Ahvaz. Rhodanese activity was assayed by the method of Sorbo in the liver, kidney, gill and intestine. The unit of enzyme activity was defined as micromoles thiocyanate formed per minute at 37 °C and pH 9.2 and enzyme activity was expressed as U/mg protein. RESULTS: Rhodanese activity was detected in all tissues studied, albeit in different amounts. Specific activities of Rhodanese (U/mg protein) in different tissues ranged from 0.135 to 0.337 in the liver, 0.113 to 0.262 in the kidney, 0.121 to 0.157 in the gill, and 0.094 to 0.162 in the intestine, respectively. CONCLUSIONS: The highest activity of Rhodanese in all four species was observed in the liver and kidney, followed by the gill and intestine. Our results suggest that Rhodanese may be functional in many physiological activities in these species which needs to be clarified in detailed.

BACKGROUND: Opioids and nitric oxide (NO) are functionally linked in the regulation of intestinal motility. OBJECTIVES: To investigate the role of NO in the opium induced bowel dysfunction in mice. METHODS: Sixty-six male mice received incrementally doses of the following treatments in six groups for 5 consecutive days: 1) Opium (0.2, 0.3, 0.4, 0.5 and 0.6mg/30g/day), 2) N-nitro-L-arginine methyl ester (L-NAME, 5,7.5,10,15 and 20mg/kg/day), 3) L-arginine (5-20mg/kg/day), 4) Opium+L-NAME, 5) Opium+L-arginine and 6) distilled water. At the end of the treatment, the abdomen was opened; some pieces of duodenal and proximal colon were taken to determine NO synthase (NOS) expression and nitrite levels, and some isolated rings from those parts of small and large intestine were prepared and transferred to the organ bath system to study intestinal motility. RT-PCR was used to determine the NOS gene expression. To determine the small intestinal transit, 30 mice in six groups, were used for oral administration of charcoal+gum in vivo. RESULTS: Opium decreased amplitude of the duodenum and ileum contractions, but increased frequency of duodenal and mid colon contractions (P<0.05). While the gene expression of inducible, neuronal and endothelial NOS was increased in colon (P<0.05), a reduced neuronal and endothelial NOS gene expression was shown in duodenum. The charcoal+gum transit was decreased in opium-treated animals compared to the control group (19.9%). However, L-arginine increased this transit while L-NAME decreased it. CONCLUSIONS: Opium induced intestinal smooth muscle spasms, which result in the decreased intestinal movements. The alterations in NOS gene expression may be a compensation mechanism against opium-induced intestinal dysfunction.

Persistence of the vaccine RH strain of Toxoplasma gondii was studied by bioassay and histologically in 14 pigs. Pigs were euthanatized 2, 4, 7, 8, 14, 15, 21, 29, 36, 42, 52, 57, and 76 days after IM inoculation with 100,000 T gondii tachyzoites. Viable T gondii tachyzoites derived from the RH strain were isolated by bioassay in mice inoculated with tissues of pigs euthanatized up to 14 days after vaccination. Except for fever, pigs vaccinated IM with the RH strain remained clinically normal. Two other pigs inoculated IV with 100,000 T gondii tachyzoites of the RH strain became ill, and 1 pig was comatose by 4 days after inoculation. These findings indicate that route of inoculation may influence the response of pigs to T gondii. To evaluate protective immunity in pigs vaccinated with the RH strain, 16 age-matched pigs were allotted to 4 groups (A-D) of 4 pigs each. Eight pigs (groups A and C) were vaccinated IM with 100,000 RH strain tachyzoites and 8 pigs (groups B and D) were nonvaccinated controls. Pigs of groups A and C were challenge-inoculated orally with a lethal dose of T gondii oocysts (100,000 oocysts) 81 days after vaccination, pigs of groups B and D were inoculated similarly 220 days after vaccination. The concentration of T gondii at 3 days after challenge inoculation of pigs vaccinated 81 days earlier was reduced 100,000-fold in mesenteric lymph nodes, compared with that in a nonvaccinated pig euthanatized at 3 days after challenge inoculation. Another nonvaccinated pig became comatose and had to be euthanatized at 7 days after challenge inoculation, numerous tachyzoites were in its mesenteric lymph nodes, intestines, and liver. The vaccinated pigs generally remained clinically after challenge inoculation with oocysts. Toxoplasma gondii was not isolated by bioassays from tissues of 5 of 8 vaccinated pigs, but was recovered from all nonvaccinated pigs. Results indicate that protective immunity persisted in pigs for at least 7 months after vaccination with the nonpersistent RH strain of T gondii.

The efficacy of a beef-based, chewable formulation of ivermectin against a mixed infection of Ancylostoma braziliense and A tubaeforme was determined in cats. Ivermectin administered orally at approximately 24 microgram/kg of body weight was 92.8% effective against adult A braziliense and 90.7% effective against adult A tubaeforme. The number of eggs per gram of feces had decreased 98.1% by 7 days after treatment. Clinical signs of hookworm disease also decreased after treatment. Location of adult parasites within the small intestine, percentage of infecting larvae that developed to the adult stage, and egg size in cats with infections of A braziliense and A tubaeforme were similar to those reported for cats with separate infections of either species.

Three commonly used keratin monoclonal antibodies (MAB)--AE1:AE3, CAM 5.2, and MAK-6--were compared with routinely used cytokeratin antibody. The expression of these antibodies was analyzed in several tissues obtained from clinically normal dogs and in a variety of neoplasms from dogs. Using appropriate enzymatic digestion, paraffin-embedded tissues processed in routine manner retained their typical keratin expression. Differentiated and poorly differentiated epithelial neoplasms, lymphomas, and melanomas were studied by use of the avidinbiotin-peroxidase technique. All 4 of the aforementioned antibodies had similar staining profiles. Of 3 anaplastic carcinomas, 2 had positive reaction to all 4 antibodies. All lymphomas, plasma cell tumors, and amelanotic melanomas had negative reaction to MAK-6, CAM 5.2, AE1:AE3, and cytokeratin MAB. Three basal cell epitheliomas had positive reaction to all 4 antibodies, whereas 1 basal cell tumor with a solid pattern had negative staining reaction. Two carcinoids had negative reaction to all markers and 1 of 2 malignant chemodectomas and 1 transitional cell carcinoma had staining reaction to only AE1:AE3 MAB. Comparing the 4 antibodies, use of AE1:AE3 MAB produced the strongest staining intensity followed by cytokeratin, MAK-6, and CAM 5.2 MAB. All 4 antibodies had low background staining. In conclusion, AE1:AE3 and MAK-6 MAB are as useful as cytokeratin MAB for identification of poorly differentiated epithelial neoplasms in dogs and cats.

Thirteen 3-week-old pigs that had been allowed to nurse for the first 16 to 18 hours after birth were orally inoculated with 1 X 10(6.5) TCID(50) of porcine rotavirus. All developed diarrhea, anorexia, and vomiting by postinoculation (PI) hour 30. These signs had abated by PI day 6. Villus blunting in the small intestine was most severe in the jejunum and ileum of pigs euthanatized between PI days 3 and 5. Villi had returned to nearly normal length by PI day 6, although fused villi were seen in a few locations in the distal portion of the jejunum and in the ileum. Virus was detected in the feces of inoculated pigs by isolation in cell cultures and by electron microscopy during the 7-day course of the experiment. There was 1 extraintestinal virus isolation from the lung of 1 pig at PI day 2. Infection and disease developed in the presence of serum-neutralizing antibody obtained by nursing seropositive sows. There was no significant change in neutralizing antibody titers in the 3-week-old pigs over the course of the experiment. In this experimental work, a model to study rotavirus infection in 3-week-old pigs has been developed.

A porcine rotavirus isolate was titrated in neonatal colostrum-fed and colostrum-deprived pigs. The stock rotavirus suspension had a titer of 10 /ml and was in its fifteenth cell culture passage in MA-104 cells. Fourteen colostrum-fed pigs were orally inoculated with dilutions of the stock virus suspension ranging from undiluted to 10-5. These pigs did not develop notable clinical signs during the 7-day experimental trial and no pathologic changes were found in intestine, liver, lung, kidney, spleen, or brain. However, rotavirus was detected in feces of the colostrum-fed pigs, using virus isolation and electron microscopic techniques. Rotavirus was also isolated from lung, brain, or spleen of 4 of 12 of these pigs. Sixteen colostrum-deprived pigs were orally inoculated with dilutions of the stock virus suspension ranging from 10-1 to 10-8. Diarrhea developed in 10 of 12 pigs that were given up to the 10-6 dilution. Seven of these 12 pigs died because of the severity of diarrhea. Pigs that died of rotavirus-induced diarrhea had severe villus loss in the jejunum and ileum. Villi of the small intestine of colostrum-deprived pigs that survived the severe diarrhea were within normal limits at the end of the 7-day trial. The colostrum-deprived pigs that were inoculated with a dilution less than 10-6 and survived past 96 hours underwent seroconversion. Rotavirus was detected by virus isolation and electron microscopy in the feces of all colostrum-deprived pigs that survived beyond 18.5 hours after inoculation. Virus was isolated from lungs, brain, or spleen of 12 of 16 colostrum-deprived pigs.

A subpopulation of purified, interepithelial lymphocytes from porcine small intestinal mucosa contained cytoplasmic granules. Toluidine blue staining revealed metachromatic granules in 13.64% (606/4,450) cells. The cells had scant organelles, a single large nucleus with obvious invagination of the nuclear membrane, and prominent chromatin. Each cell contained 1 to 10 cytoplasmic membrane-bound granules, 0.6 to 1.5 microns in diameter. These findings indicated that the granular mucosal lymphocytes are related morphologically to mucosal mast cells. The presence of serotonin in the granules, confirmed by the serotonin releasing test, provided functional evidence that granular mucosal lymphocytes are related to mucosal mast cells.

Uptake of ferritin by M cells in follicle-associated epithelium at various sites in the small and large intestines was examined in 4 healthy 5-week-old pigs by use of electron microscopy. A 2.5% solution of ferritin in saline was injected into ligated loops of the jejunum and ileum containing aggregations of lymphoid follicles (Peyer's patches), as well as into intestinal loops containing lymphoglandular complexes at the ileocecal junction, in the central colonic flexure, and in the rectum. As negative control, saline solution was injected into loops at identical localizations. After an exposure period of 2 hours, uptake of ferritin by M cells, but not by enteroabsorptive cells of the small and large intestines, was observed. Numbers of M cells with ferritin and total M cells were counted and the percentage was calculated. Total number of M cells was highest in lymphoglandular complexes in the rectum and lowest on domes of the ileal Peyer's patch. High numbers of M cells with ferritin were found on domes of the jejunal Peyer's patch, and in lymphoglandular complexes at the ileoceral entrance and in the rectum. Only a few M cells on domes of the ileal Peyer's patch and in lymphoglandular complexes in the central colonic flexure contained ferritin. The percentage of M cells with internalized ferritin was similar on domes of the ileal Peyer's patch, and in lymphoglandular complexes at the ileocecal junction and in the rectum. It was higher on domes of the jejunal Peyer's patches and lower in lymphoglandular complexes of the central colonic flexure. Ferritin was found in the apical tubulovesicular system, multivesicular bodies, and a few vacuoles in the central area of M cells. Ferritin was exocytosed into the lateral intercellular spaces next to M cells. Uptake of ferritin by intraepithelial cells in the follicle-associated epithelium could not be documented, but ferritin was present in vesicles of subepithelial macrophages.

Diamine oxidase (DAO), an enzyme of small intestinal origin, is released from mucosal storage sites by IV administration of heparin, to yield the plasma postheparin DAO (PHD) curve. The PHD curve is diminished when mucosal surface area is lost, and baseline (without heparin) plasma DAO activity increases when mucosal storage sites are damaged. Plasma DAO activity was measured after 2 doses of heparin were administered Iv in healthy, conscious horses. In anesthetized horses, the PHD curve was studied: during sham small intestinal surgery, and during venous strangulation obstruction (VSO) of the distal 50% of the small intestine. In a third group of anesthetized horses, baseline plasma DAO activity (without heparin) was measured during vso of the distal 50% of the small intestine for 90 minutes, followed by reperfusion for 90 minutes. Postheparin plasma DAO curves in conscious horses were similar to those reported in other species Horses with VSO had a similar PHD curve as did sham-operated controls at all times, except at 15 minutes, when plasma DAO activity was significantly (P < 0.05) greater in the vso group. Horses with VSO and reperfusion had no change in baseline plasma DAO activity throughout the study. Peritoneal fluid DAO activity remained low throughout the study, but increased slightly in horses with VSO that received heparin, possibly because of DAO from extravasated blood in the peritoneal fluid. Results indicated that the plasma DAO response to IV administered heparin in horses is similar to that in other mammals, but, unlike other species, baseline and postheparin DAO activities did not change as expected after small intestinal vascular obstruction and mucosal injury. There may be additional sources of DAO in horses, the type of injury induced was not of sufficient magnitude to affect storage sites of DAO, or the circulatory changes induced by vso might have altered tissue delivery of heparin.