Fetal and neonatal goiter in cynomolgus monkeys following administration of the antithyroid drug thiamazole at high doses to dams during pregnancy

To evaluate morphologic alterations in the thyroid gland in the second generation in cynomolgus monkeys, pregnant dams were exposed to high doses of thiamazole. In Experiment A, dams received thiamazole intragastrically via a nasogastric catheter from gestation day (GD) 50 to GD 150 or on the day before delivery. Initially, the dose level was 20 mg/kg/day (10 mg/kg twice daily); however, the dose level was subsequently decreased to 5 mg/kg/day (2.5 mg/kg twice daily), since deteriorated general conditions were observed in two dams. Six out of seven neonates died on the day of birth. The cause of neonatal death was tracheal compression and suffocation from goiter. The transplacental exposure to thiamazole affected the fetal thyroid glands and induced goiter in all neonates. The surviving neonate was necropsied 767 days after discontinuation of thiamazole exposure and showed reversibility of the induced changes. In Experiment B, dams were intragastrically administered thiamazole at 5 mg/kg/day (2.5 mg/kg twice daily) for treatment periods from GDs 51 to 70, 71 to 90, 91 to 110, 111 to 130 and 131 to 150. All fetuses showed enlarged thyroid glands but were viable. Histopathologically, hypertrophy and/or hyperplastic appearance of the follicular epithelium of the thyroid gland was observed at the end of each treatment period. The most active appearance of the follicular epithelium, consisting of crowded pedunculated structure, was demonstrated at end of the treatment period from GD 131 to 150. This is the first report on the morphology of fetal and neonatal goiter in the cynomolgus monkey.     

Multi-locus sequence analysis reveals host specific association between Bartonella washoensis and squirrels

To clarify phylogenetic relationships and genetic diversity among Bartonella washoensis strains obtained from squirrels, multi-locus sequence analysis (MLSA) with the 16S rRNA, ftsZ, gltA, groEL, ribC, and rpoB genes was applied for 20 strains of B. washoensis isolated from five genera of squirrels (Tamias, Tamiasciurus, Glaucomys, Sciurus, and Spermophilus) within the family Sciuridae. Sequence similarities in the concatenated sequences of B. washoensis strains from squirrels of different genera ranged from 94.7% (Sciurus vs. Spermophilus) to 98.4% (Tamiasciurus vs. Glaucomys). Phylogenetic trees based on the concatenated sequences revealed that B. washoensis strains formed five distinct clades and each clade correlated with the genus of squirrel from which the strains were originally obtained. The discrimination was supported by 100% bootstrap values and posterior probabilities, respectively. These results suggest that B. washoensis strains may have co-speciated with their squirrel hosts and provide new insights into the application of the MLSA to identify sources of B. washoensis infection with accuracy.     

Evaluation of feeding value of forage soybean silage as a substitute for wheat bran in sheep

Twelve sheep were used in a 3 × 3 Latin square design to evaluate the effect of substituting wheat bran with forage soybean silage in the diet on apparent digestibility and nitrogen balance. Forage soybean (Glycine max (L.) Merrill) was cultivated in a no‐till, no‐herbicide cropping system with Italian ryegrass (Lolium multiflorum Lam.) as a living mulch. Forage soybean and Italian ryegrass were wilted and ensiled in round bales without additives, respectively. The experimental diets were based on corn silage supplemented with protein sources (tow silages or wheat bran). The crude protein and the acid detergent insoluble protein contents of forage soybean silage were the highest among the protein sources. The apparent digestibility of crude protein and the nitrogen balance did not significantly differ among the diets. In addition, the phytoestrogen content of forage soybean silage was below the level at which animal reproductive performance would be negatively affected. These results suggest that forage soybean silage has comparable feeding value to wheat bran, and can be given at an inclusion level of 17% (dry matter basis) as an alternative protein source to wheat bran without adverse effects on digestion or nitrogen balance in sheep fed a corn silage‐based diet.     

Prevalence of zoonotic Bartonella species among rodents and shrews in Thailand

We investigated the prevalence of Bartonella species in 10 rodent and one shrew species in Thailand. From February 2008 to May 2010, a total of 375 small animals were captured in 9 provinces in Thailand. Bartonella strains were isolated from 57 rodents (54 from Rattus species and 3 from Bandicota indica) and one shrew (Suncus murinus) in 7 of the 9 provinces, and identified to the species level. Sequence analysis of the citrate synthase and RNA polymerase β subunit genes identified the 58 isolates from each Bartonella-positive animal as B. tribocorum in 27 (46.6%) animals, B. rattimassiliensis in 17 (29.3%) animals, B. elizabethae in 10 (17.2%) animals and B. queenslandensis in 4 (6.9%) animals. R. norvegicus, R. rattus, and Suncus murinus carried B. elizabethae, which causes endocarditis in humans. The prevalence of Bartonella bacteremic animals by province was 42.9% of the animals collected in Phang Nga, 26.8% in Chiang Rai, 20.4% in Sa Kaeo, 16.7% in Nakhon Si Thammarat, 12.0% in Surat Thani, 9.1% in Mae Hong Son and Loei Provinces.     

CEP peptide induces susceptibility of Arabidopsis thaliana to non-adapted pathogens

CEP peptide was synthesized and tested for induction of disease susceptibility using Arabidopsis Col-0. When Colletotrichum tropicale was used as a non-adapted fungal pathogen, the conidia germinated to form hyphal-like structures, which successfully penetrated epidermis, eventually causing disease symptoms. In such case, PEN2-, but not PEN3-dependent resistance was likely suppressed by CEP peptide. Similarly, the CEP peptide-mediated disease susceptibility was also effective to a non-adapted bacterial pathogen. Notably, such induced susceptibility was also evident on Arabidopsis mutants lacking the previously identified receptors, suggesting that the CEP peptide modulates Arabidopsis immunity through an unidentified receptor(s).

     

Identification and Biochemical Characterization of Halisulfate 3 and Suvanine as Novel Inhibitors of Hepatitis C Virus NS3 Helicase from a Marine Sponge

Hepatitis C virus (HCV) is an important etiological agent that is responsible for the development of chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. HCV nonstructural protein 3 (NS3) helicase is a possible target for novel drug development due to its essential role in viral replication. In this study, we identified halisulfate 3 (hal3) and suvanine as novel NS3 helicase inhibitors, with IC₅₀ values of 4 and 3 µM, respectively, from a marine sponge by screening extracts of marine organisms. Both hal3 and suvanine inhibited the ATPase, RNA binding, and serine protease activities of NS3 helicase with IC₅₀ values of 8, 8, and 14 µM, and 7, 3, and 34 µM, respectively. However, the dengue virus (DENV) NS3 helicase, which shares a catalytic core (consisting mainly of ATPase and RNA binding sites) with HCV NS3 helicase, was not inhibited by hal3 and suvanine, even at concentrations of 100 µM. Therefore, we conclude that hal3 and suvanine specifically inhibit HCV NS3 helicase via an interaction with an allosteric site in NS3 rather than binding to the catalytic core. This led to the inhibition of all NS3 activities, presumably by inducing conformational changes.     

Seroprevalence of Toxoplasma gondii antibodies in stray cats and dogs in the Bangkok metropolitan area, Thailand

Cats and dogs are the most popular pet animals worldwide. Cats are the natural reservoir of Toxoplasma gondii and excrete the resistant oocyst to environments. On the other hands, dogs play a role in the mechanical transmission of the parasite. Stray cats and dogs in the Bangkok metropolitan area are becoming a public concern because there is a considerable increase in their number annually. These facts indicate the risk of mechanically spreading zoonoses including toxoplasmosis to humans since human acquire the infection from infected mammals, either directly or indirectly. In the present study, the presence of T. gondii antibodies was examined in 592 cats and 427 dogs from October 2001 to September 2002 by using a latex agglutination test. T. gondii antibodies were detected in 65 (11.0%) of the 592 cats and 40 (9.4%) of the 427 dogs. The antibody titers in the positive animals ranged from 1:64 to 1:2048. Seroprevalence was significantly higher in female cats than in male cats. The present study suggested that T. gondii was widespread in the stray animals in the Bangkok metropolitan area; therefore, it is essential to control the number of stray cats and dogs in order to reduce the transmission of toxoplasmosis to animals and humans.     

Simultaneous determination of antimony and bismuth in soils by continuous hydride generation—atomic absorption spectrometry

A method for the simultaneous determination of Sb and Bi in soils using an automated hydride generation system with an electrically heated quartz cell atomizer, and an atomic absorption spectrophotometer is described. Five g of the air-dried soil was placed in a 100 mL Erlenmyer flask fitted with a Liebig condenser and boiled gently with 20 mL 6 M HCl for 1 hr on a sand bath. The extract was then filtered into a 100 mL volumetric flask. The two elements in the sample solution (<30 mL) placed in a 50 mL volumetric flask are prereduced by potassium iodide and ascorbic acid solutions. Reduction of Sb and Bi ions to stibine and bismuthine is obtained by sodium tetrahydroborate solution. Detection limits (S/N = 2) are 7 ng g^?1 dry soil for Sb and 0.7 ng g^?1 dry soil for Bi. The effects of 19 diverse ions on the absorbances of Sb and Bi were investigasted, and it was found that the errors by these ions will be within 5% in the ordinary unpolluted or even polluted soils. In order to examine the accuracy of the proposed method, Sb and Bi in the four Canadian reference soils and NBS-4355 from the National Bureau of Standards in the USA were determined. The values obtained in this work are close to the recommended or the not-certified values. The amounts of Sb, Bi and the related elements in the soils near a smelter were also analyzed.     

Quantification of genetically modified soybean by quenching probe polymerase chain reaction

Quenching probe (QProbe) polymerase chain reaction (PCR) is a simple and cost-effective real-time PCR assay in comparison with other real-time PCR assays such as the TaqMan assay. We used QProbe-PCR to quantify genetically modified (GM) soybean (Roundup Ready soybean). We designed event-specific QProbes for Le1 (soy endogenous gene) and RRS (recombinant gene), and we quantified certified reference materials containing 0.1, 0.5, 1, 2, and 5% GM soybean. The TaqMan assay was also applied to the same samples, and the results were compared. The accuracy of QProbe-PCR was similar to that of TaqMan assay. When GM soybean content was 0.5% or more, the relative standard deviations of QProbe-PCR were less than 20%. QProbe-PCR is sensitive enough to monitor labeling systems and has acceptable levels of accuracy and precision.