Lactobacillus rhamnosus GG SpaC pilin subunit binds to the carbohydrate moieties of intestinal glycoconjugates

Lactobacillus rhamnosus GG (LGG) is a well‐established probiotic strain. The beneficial properties of this strain are partially dependent on its prolonged residence in the gastrointestinal tract, and are likely influenced by its adhesion to the intestinal mucosa. The pilin SpaC subunit, located within the Spa pili structure, is the most well studied LGG adhesion factor. However, the binding epitopes of SpaC remain largely unknown. The aim of this study was to evaluate the binding properties of SpaC to the carbohydrate moieties of intestinal glycoconjugates using a recombinant SpaC protein. In a competitive enzyme‐linked immunosorbent assay, SpaC binding was markedly reduced by addition of purified mucin and the mucin oligosaccharide fraction. Histochemical staining revealed that the binding of SpaC was drastically reduced by periodic acid treatment. Moreover, in the surface plasmon resonance‐based Biacore assay, SpaC bound strongly to the carbohydrate moieties containing β‐galactoside at the non‐reducing terminus of glycolipids. We here provide the first demonstration that SpaC binds to the oligosaccharide chains of mucins, and that the carbohydrate moieties containing β‐galactoside at the non‐reducing termini of glycoconjugates play a crucial role in this binding. Our results demonstrate the importance of carbohydrates of SpaC for mucus interactions.     

Intrapancreatic accessory spleen mimicking pancreatic insulinoma with intrapancreatic metastasis in a cat

An 11-year-old neutered male Domestic Shorthair cat presented with a 3-month history of hypoglycemia, two episodes of seizure, and intermittent tick-like signs. Serum biochemistry revealed severe hypoglycemia associated with high insulin concentrations. Dynamic abdominal computed tomography (CT) indicated two pancreatic masses, which were enhanced most during the late arterial phase but had different degrees and variations of attenuation. Partial pancreatectomy was performed. Histopathology and immunohistochemistry confirmed that one mass was an insulinoma and the other was an ectopic splenic tissue, consistent with the differences in imaging findings. When an intrapancreatic lesion with hyper-attenuation on dynamic abdominal CT is detected, not only insulinoma or metastasis of malignancies but also intrapancreatic accessory spleen (IPAS) should be considered as differential diagnoses.     

Effects of tris(1,3-dichloro-2-propyl) phosphate on epididymal sperm parameters in adult male rats

Tris(1,3-dichloro-2-propyl) phosphate (TDCIPP) is widely used as a flame retardant and is known to exhibit anti-androgenic effects in vitro and in vivo. To assess the reproductive toxicity potency of TDCIPP, we investigated the effects of 7 days of TDCIPP oral administration on epididymal sperm motion and concentration in adult male Wistar–Imamichi rats. Thirty-five days after the final administration, sperm parameters were evaluated by computer-assisted sperm analysis. Results showed that sperm swimming progression and vigor and sperm concentration in TDCIPP-treated rats were unexpectedly higher than those in control rats. TDCIPP did not significantly affect the percentage of motile sperms or sperm swimming pattern. These results contribute to the understanding of the biological effects of TDCIPP.     

Validation and application of real-time polymerase chain reaction assays for representative rumen bacteria

Real‐time polymerase chain reaction (PCR) assays for 11 representative rumen bacterial species were validated. The sensitivity was tested by using the serially diluted target 16S rDNA from respective bacterial species. The recovery of the target DNA and the assay reproducibility were determined using DNA from rumen fluid spiked with different quantities of the target. Minimum detection levels for the target were 10–100 copies in pure culture. The recovery of the added target ranged from 82.4 to 116.6%. The intra‐ and inter‐assay variations of each assay were <9.4 and <12.6%, respectively. Therefore, the real‐time PCR assays evaluated in the present study are considered to be sufficiently reliable for monitoring all 11 bacterial species in the rumen. The assays were then applied to the monitoring of the bacterial species attached to ruminally incubated rice straw. Among the monitored fibrolytic species, Fibrobacter succinogenes was found to be the most dominant, accounting for 2.61% of total bacteria after 24 h incubation. Selenomonas ruminantium and Streptococcus bovis, non‐fibrolytics, were detected on the rice straw at 8.96% and 1.16% of total bacteria, respectively. Such high levels of non‐fibrolytics on the plant fiber suggest a synergistic relationship between fibrolytics and non‐fibrolytics.     

Adrenocorticotropic hormone-induced secretion of cortisol in goats is inhibited by androgen

In a previous study, it was found that there are sex differences in goats with respect to the levels of cortisol secretion induced by transportation stress. We also found that treatment of castrated male goats with dihydrotestosterone (DHT) suppressed the increase in plasma cortisol concentration following transportation, but did not suppress the secretion of adrenocorticotropic hormone (ACTH). This suggests that androgen might block ACTH ‐ induced cortisol secretion. In order to examine this hypothesis, the effects of androgen on ACTH‐induced cortisol secretion in goats were investigated. First, castrated male goats were treated with testosterone (T), DHT or cholesterol (cho) for 21–25 days. Cho was used as a control for T and DHT treatment. Then, plasma cortisol concentrations were compared among the hormonal treatments after ACTH injection. Subsequently, the distribution of androgen receptors in the caprine adrenal gland was investigated. There were no differences in the basal cortisol concentrations among the hormonal treatments. However, plasma cortisol concentrations after ACTH injection were significantly lower in T ‐ and DHT ‐ treated goats than in cho ‐ treated goats. Androgen receptors were present in 60% of the cells in the zonae fasciculata and reticularis of the adrenal cortex, the regions that secrete glucocorticoids. These results suggest that androgen may act directly on the adrenal cortex to suppress cortisol secretion induced by ACTH.     

Combined effects of clenbuterol and various concentrations of protein on performance of broiler chickens

1. The effects of the beta-adrenergic agonist, clenbuterol (1 mg/kg diet) on the growth and muscle composition in female broiler chickens (14 to 32 d of age) fed on diets containing various concentrations of protein (220, 240 or 260 g protein/kg) were examined. 2. Body weight gain over the 18 d period increased linearly with increasing protein intake. The rate of gain was significantly higher in clenbuterol-treated chickens than in control birds. 3. Dietary clenbuterol increased thigh muscle weight and protein concentration of breast and thigh muscle, regardless of dietary protein content. Protein/DNA ratio in thigh muscle was enhanced by clenbuterol feeding, and the magnitude of difference of the ratio was higher in chickens fed on the 240 and 260 g CP/kg diets than in those fed the 220 g CP/kg diet. 4. It was concluded that clenbuterol-treated chickens require increased dietary protein to maintain maximal growth, and that increased protein consumption is an important factor in improving growth in clenbuterolfed broilers.