Inhibition of juvenile hormone biosynthesis in the tobacco hornworm, Manduca sexta, by a bisthiolcarbamate juvenoid and related compounds

Biosynthesis of juvenile hormone in the tobacco hornworm, Manduca sexta, is inhibited by the bisthiolcarbamate juvenoid N-ethyl-1,2-bis(isobutylthiolcarbamoyl)ethane both in vitro and in vivo. In vitro an extremely steep dose-response curve was obtained with an ID₅₀ value of 6 × 10^?6M. However, in vivo topical treatment with the compound resulted in mild JH antagonistic symptoms, suggesting rapid metabolism of the compound. In agreement with results from metabolic studies performed on plants and in mammals, sulfoxidation of the thiocarbamate S-(4-chlorobenzyl)N,N-diethylthiocarbamate resulted in an enhanced inhibitory effect on JH biosynthesis in vitro. This suggests that the corresponding thiocarbamate sulfoxides may act as intermediates in carbomylating critical thiol sites important in the terpenoid biosynthesis pathway. Furthermore, this study shows that these prototype compounds are interesting tools for further investigation of chemical inhibition of JH biosynthesis in insects.     

The bovine neutrophil: Structure and function in blood and milk

Migration of polymorphonuclear neutrophil leukocytes (PMN) into the mammary gland provide the first line of defense against invading mastitis pathogens. Bacteria release potent toxins that activate white blood cells and epithelial cells in the mammary gland to secrete cytokines that recruit PMN that function as phagocytes at the site of infection. While freshly migrated PMN are active phagocytes, continued exposure of PMN to inhibitory factors in milk such as fat globules and casein, leads to altered PMN morphology and reduced phagocytosis. In the course of phagocytosing and destroying invading pathogens, PMN release chemicals that not only kill the pathogens but that also cause injury to the delicate lining of the mammary gland. This will result in permanent scarring and reduced numbers of milk secretory cells. The life span of PMN is limited by the onset of apoptosis. To minimize damage to mammary tissue, PMN undergo a specialized process of programmed cell death known as apoptosis. Macrophages quickly engulf and phagocytose apoptotic PMN, thereby minimizing the release of PMN granular contents that are damaging to tissue. The PMN possess an array of cell surface receptors that allow them to adhere and migrate through endothelium and to recognize and phagocytose bacteria. One receptor found on phagocytes that is receiving considerable attention in the control of infections by Gram-negative bacteria is CD14. Binding of lipopolysaccharide (LPS) to membrane bound CD14 causes release of tumor necrosis factor-alpha and sepsis. Binding of LPS to soluble CD14 shed from CD14-bearing cells results in neutralization of LPS and rapid recruitment of PMN to the site of infection. Recent advances in the fields of genomics and proteomics should greatly enhance our understanding of the PMN role in controlling intramammary infections in ruminants. Further, manipulation of PMN, through either recombinant proteins such as soluble CD14 that enhance PMN response or agents that mediate PMN apoptosis, may serve as novel therapeutics for the treatment of mastitis.     

Flow cytometric assessment of ethylene glycol monoethyl ether on spermatogenesis in rats

The effects of ethylene glycol monoethyl ether (EGEE) on testicular cell populations in rats were investigated by a flow cytometric method. Rats were administered by gavage with EGEE at the various doses of 0 (saline alone), 100, 200, 400, and 800 mg/kg body weight/day for 4 weeks. The treatment of EGEE caused decreases in the weight of testis and epididymis and in the number of testicular cells. Histopathologically, exfoliation of germ cells into the tubular lumen was observed at the doses of above 200 mg/kg. The treatment of EGEE at the dose of 400 mg/kg caused moderate testicular degeneration. A significant depletion of haploid cells and a disproportionate ratio of diploid and tetraploid cells were observed as determined by flow cytometric analysis. These results indicate that the toxic effect of EGEE on the male reproductive system may be strongly associated with the disproportion of testicular germ cells.     

Effect of intravenous mannitol upon the resistive index in complete unilateral renal obstruction in dogs

Some studies have shown that relative to baseline, the renal resistive index (RI) remains unchanged in nonobstructed kidneys and increases in obstructed kidneys after administration of furosemide. To our knowledge, the effect of mannitol administration on the renal RI of dogs has not been reported. We evaluated the renal RI in 16 kidneys in 8 young adult dogs after administration of mannitol. The mean RI decreased significantly from baseline (P < .01). Additionally, left complete ureteral obstruction wasinduced in 5 dogs. Evaluation by Doppler ultrasonography was performed for 5 days. On the 5th day, Doppler examination was repeated at 30 and 60 minutes after administration of mannitol to obstructed dogs. After induction of left ureteral obstruction, the RI of the left kidney increased significantly over 5 consecutive days. Administration of mannitol decreased the RI in the nonobstructed contralateral kidneys, and thus the RI difference between obstructed and nonobstructed kidneys was increased above normal (P < .001). In conclusion, administration of mannitol may be useful as another diuretic agent to identify unilateral ureteral obstruction on Doppler sonographic examination.     

Histochemical detection of glycoconjugates in the male reproductive system of the horse

Lectins are glycoproteins of plant and animal origin that have the ability to bind specific carbohydrate residues of cell glycoconjugates, particularly in terminal positions. In this study, the binding of lectins, Dolichos biflorus agglutinin (DBA), soybean agglutinin (SBA), Bandeiraea simplicifolia BS-1 (isolectin B4), Triticum vulgaris (WGA), Arachis hypogaea (PNA), and Ulex europaeus (UEA-I), was studied in the reproductive systems of male thoroughbred horses.DBA was detected in the stereocilia of the caput and corpus epididymis, and in the vas deferens. It was weakly detected in connective tissue of the corpus epididymis. Strong SBA staining was seen in epithelial cells in the testis, stereocilia of the corpus and cauda epididymis, and in the vas deferens. There were intense positive reactions for isolectin B4 in interstitial cells in all tissue and serosa of the vas deferens. PNA staining was seen only in stereocilia in the caput and corpus epididymis, and in the vas deferens. Strong WGA staining was seen throughout the testis, except in Sertoli cells, stereocilia, and connective tissue. UEA-I was detected in secondary spermatids, stereocilia, and epithelial cells of the cauda epididymis. These results show that degenerating cells in the testis, epididymal tubules, and vas deferens have differential affinities for lectins, and suggest that lectins play a role in the reproductive system of the horse. The heterogeneity of the lectin staining pattern in the reproductive tubules of adult horses suggests that the carbohydrate composition of each cell type is region specific.     

Detection of Campylobacter jejuni strains in the water lines of a commercial broiler house and their relationship to the strains that colonized the chickens

Campylobacter jejuni is frequently present in the intestinal tract of commercial broiler chickens, and their drinking water has been proposed to be an initial source of bacteria for newly hatched chicks. We studied three sequential commercial broiler flocks raised in a house from which we had cultured C. jejuni from the nipple waters prior to placement of the first flock. Campylobacter cells were detected by immunofluorescence in the biofilm of the drinking nipples during the weeks when the flock was colonized with C. jejuni but not during weeks when the birds were negative. Campylobacter jejuni was isolated from the drinking water during the growth of the first flock and was present in the birds from all three flocks. Randomly amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR) typing with primer OPA11 indicated that seven distinct strains were present within the broiler house. One strain found in drinking water was similar to a strain found in birds in the second flock; however, RAPD-PCR with primer HLW85 showed that the strains were not identical. These results suggest that although the watering system is a potential source of C. jejuni in broiler flocks, the waterborne strain in this study was not detected in the birds.     

Effects of dietary essential oil components on growth performance,digestive enzymes and lipid metabolism in female broiler chickens

1. The present experiment was conducted to describe the effects of thymol, cinnamaldehyde and a commercial preparation of essential oil components (CRINA Poultry), in female broilers. Feed and water were provided for ad libitum consumption. 2. Feed intake, weight gain and feed:gain ratio were not different among the treatments. Water intake was significantly lowered by cinnamaldehyde. Relative liver weight (g/100 g of body weight) was highest in birds given thymol, but this was seen only at the age of 21 d and not at 40 d. Patterns of digestive enzymes in pancreatic tissue were similar for the 4 treatments. 3. Amylase activity in intestinal digesta was highest in chickens given CRINA Poultry for 21 d, but the effect had disappeared after 40 d. Ileal digestibility coefficients for starch and protein were high and identical for all treatments. 4. Fatty acid composition of diet was reflected in that of adipose tissue. Plasma lipid concentrations were not changed by any dietary treatment. 5. Thus, the present results show no effect of essential oil constituents on growth performance in female broiler chickens, but it cannot be excluded that positive effects would have been observed under less hygienic environmental conditions or when using a less digestible diet.     

Effects of dietary electrolyte balance on the chemistry of blood and urine in lactating sows and sow litter performance

One hundred fifty-three sows (average parity of 2.2) were used to determine the effects of dietary electrolyte balance (calculated as mEq/kg of diet for Na + K - Cl) on sows and their litters during lactation. The sows were fed corn-soybean meal-based diets (1.0% lysine, 1.0% valine, 0.95% Ca, and 0.80% P; as-fed basis) starting on d 109 of gestation and throughout the 21-d lactation experiment. Dietary electrolyte balance (dEB) was 0, 100, 200, 350, and 500 mEq/kg (as-fed basis), well above and below the dEB of 185 mEq/kg found in a simple corn-soybean meal-based lactation diet. To achieve the desired dEB, diets had the following: 1) 1.8% HCl (6 N) and 1.06% CaCl2, 2) 1.0% CaCl2, 3) 0.04% NaHCO3, 4) 1.29% NaHCO3, and 5) 2.54% NaHCO3 (as-fed basis). Increasing dEB increased blood pH (linear and quadratic effects, P < 0.001), partial pressure of carbon dioxide (linear effect, P < 0.001), HCO3- concentration (linear and quadratic effects, P < 0.001), and blood base excess (linear and quadratic effects, P < 0.001). However, increased dEB resulted in lower blood concentrations of K (linear and quadratic effects, P < 0.04), Cl (linear and quadratic effects, P < 0.001), and ionized Ca (linear and quadratic effects, P < 0.001). Changing dEB did not affect ADFI; water usage, litter weight gain; sow weight change; sow backfat change; percentages of CP, lactose, and fat in the milk; percentage of sows returning to estrus; days to estrus; and number of pigs born alive in the subsequent litter (P = 0.06). However, piglet survivability to d 10 and overall was greatest with the lower dEB treatments (linear effect, P < 0.05). The pH (linear and quadratic effects, P < 0.001) and colony forming units of total bacteria (linear effect, P < 0.03) in the urine increased as dEB of the diet was increased. In conclusion, dEB had pronounced effects on the physiological status of sows and decreasing dEB below that in a simple corn-soybean meal-based diet decreased bacterial counts in the urine and increased piglet survivability. However, milk composition, sow and litter weights at weaning, and subsequent rebreeding performance of the sows were not affected by dEB.     

Pax-7 immunoreactivity in the post-natal chicken central nervous system

In this immunocytochemical study on the constitutive expression of Pax-7 protein in the postnatal chicken brain, Pax-7 showed region and cell type specific expression. In the optic tectum, only cells in grey matter showed positive immunoreactivities (IRs), whereas those in the white matters did not show any IRs. In thalamic nuclei and several pontine nuclei, we also localized Pax-7 positive IRs. On the contrary, in the cerebellum, Pax-7 was mainly localized within the Bergmann glia, whereas Purkinje cells did not show any IRs. In double immunolabelling studies, most of the Pax-7 IRs did not originate from neuroglial cells such as oligodendrocytes, microglia or astrocytes, but from neurons, with the exception of Bergmann glia in the cerebellum. The presence of Pax-7 IRs in the adult chicken brain could suggest that Pax-7 might play a role in maintaining normal physiological function in some postnatal chicken brain cells.     

Characterization of monoclonal antibodies against avian reovirus S1133 protein sigmaA synthesized in Escherichia coli

Monoclonal antibodies (MAbs) were prepared against avian reovirus S1133 protein sigmaA (esigmaA) synthesized in Escherichia coli. MAbs were characterized and used to develop a diagnostic test. Ten MAbs were selected for competitive binding assay following coupling with horseradish peroxidase. The results indicated that these MAbs delineated two epitopes I and II of esigmaA. An immuno-dot binding assay was used to detect the effect of denaturation on antibody recognition of the epitopes. All MAbs bound to esigmaA in its native form. After denaturation by boiling in SDS and 2-mercaptoethanol, the binding of MAbs recognizing epitope I was fully abolished. However, the reactivity of MAbs recognizing epitope II was not affected. MAbs 31 and 32, recognizing epitopes I and II, respectively, were selected for the cross-reactivity to heterologous reovirus strains. The results suggest that the two epitopes are highly conserved among these virus strains. A MAb capture enzyme-linked immunosorbent assay (ELISA) procedure was developed using MAbs 32 and 31 to detect reovirus protein sigmaA in samples from tendon tissues of infected bird and chicken embryo fibroblast (CEF) cell cultures. Avian reovirus sigmaA antigens in tendon specimens were detected from the inoculated birds as early as 2 days post-inoculation (PI), approximated a peak at 7 days PI, and maintained this until 16 days PI, then decreased gradually. A clear difference in absorbance values between the tendon samples of the avian reovirus- and mock-infected birds is obtained. Positive results were also obtained from avian reovirus-infected CEF and from the tendon tissues of naturally infected broilers. These results indicated that the MAb capture ELISA is a useful methods for the detection of avian reovirus from chickens suspected to have avian reovirus infections.