The combined effect of clothianidin and environmental stress on the behavioral and reproductive function in male mice

Neonicotinoids, some of the most widely used pesticides in the world, act as agonists to the nicotinic acetylcholine receptors (nAChRs) of insects, resulting in death from abnormal excitability. Neonicotinoids unexpectedly became a major topic as a compelling cause of honeybee colony collapse disorder, which is damaging crop production that requires pollination worldwide. Mammal nAChRs appear to have a certain affinity for neonicotinoids with lower levels than those of insects; there is thus rising concern about unpredictable adverse effects of neonicotinoids on vertebrates. We hypothesized that the effects of neonicotinoids would be enhanced under a chronic stressed condition, which is known to alter the expression of targets of neonicotinoids, i.e., neuronal nAChRs. We performed immunohistochemical and behavioral analyses in male mice actively administered a neonicotinoid, clothianidin (CTD; 0, 10, 50 and 250 mg/kg/day), for 4 weeks under an unpredictable chronic stress procedure. Vacuolated seminiferous epithelia and a decrease in the immunoreactivity of the antioxidant enzyme glutathione peroxidase 4 were observed in the testes of the CTD+stress mice. In an open field test, although the locomotor activities were not affected, the anxiety-like behaviors of the mice were elevated by both CTD and stress. The present study demonstrates that the behavioral and reproductive effects of CTD become more serious in combination with environmental stress, which may reflect our actual situation of multiple exposure.     

Cassava starch factory residues in the diet of slow-growing broilers

The objective of this study was to evaluate the effect of inclusion of dehydrated cassava starch residue (DCSR) on the performance, gastrointestinal tract characteristics and carcass traits of ISA Label JA57 slow-growing broilers. A total of 510 broilers at 21 were distributed in a randomized experimental design with 5 treatments (2, 4, 6, 8, and 10 % DCSR inclusion) and a control group, 5 replicates, and 17 birds per experimental unit. The DCSR inclusion from 21 to 49 days of age negatively influenced (P?P?>?0.05) feed conversion in the broilers with increasing of DCSR inclusion. From 21 to 79 days, DCSR inclusion impaired (P?P?P?P?>?0.05) the other parameters of carcass quality and yield, cuts, and percentage of abdominal fat. In conclusion, DCSR inclusion levels above 2 % compromised broiler performance.     

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.     

Isolation of a novel mutant gene for soil-surface rooting in rice (Oryza sativa L.)

Background

Root system architecture is an important trait affecting the uptake of nutrients and water by crops. Shallower root systems preferentially take up nutrients from the topsoil and help avoid unfavorable environments in deeper soil layers. We have found a soil-surface rooting mutant from an M₂ population that was regenerated from seed calli of a japonica rice cultivar, Nipponbare. In this study, we examined the genetic and physiological characteristics of this mutant.

Results

The primary roots of the mutant showed no gravitropic response from the seedling stage on, whereas the gravitropic response of the shoots was normal. Segregation analyses by using an F₂ population derived from a cross between the soil-surface rooting mutant and wild-type Nipponbare indicated that the trait was controlled by a single recessive gene, designated as sor1. Fine mapping by using an F₂ population derived from a cross between the mutant and an indica rice cultivar, Kasalath, revealed that sor1 was located within a 136-kb region between the simple sequence repeat markers RM16254 and 2935-6 on the terminal region of the short arm of chromosome 4, where 13 putative open reading frames (ORFs) were found. We sequenced these ORFs and detected a 33-bp deletion in one of them, Os04g0101800. Transgenic plants of the mutant transformed with the genomic fragment carrying the Os04g0101800 sequence from Nipponbare showed normal gravitropic responses and no soil-surface rooting.

Conclusion

These results suggest that sor1, a rice mutant causing soil-surface rooting and altered root gravitropic response, is allelic to Os04g0101800, and that a 33-bp deletion in the coding region of this gene causes the mutant phenotypes.     

rDNA-based characterization of a new binucleate Rhizoctonia spp. causing root rot on kale in Brazil

In this paper we present the first report of the occurrence of a binucleate Rhizoctonia spp. causing hypocotyl and root rot in kale in Brazil. Rhizoctonia spp. were isolated from kale (Brassica oleracea var. acephala) with symptoms of hypocotyl and root rot. The isolates, characterized as binucleate Rhizoctonia spp., did not show an anastomosis reaction with any of the binucleate Rhizoctonia spp. testers used. The pathogenicity of the isolates was tested under greenhouse conditions; all isolates were pathogenic and showed different symptom severities on kale. The ITS-5.8S rDNA sequences of kale isolates and 50 testers (25 binucleate Rhizoctonia spp. and 25 Rhizoctonia solani) were compared in order to characterize the genetic identity of Rhizoctonia spp. infecting kale. The kale isolates showed genetic identities ranging from 99.3 to 99.8% and were phylogenetically closely related to CAG 7 (AF354084), with identities of 98.5 and 98.7%. It is suggested that the binucleate Rhizoctonia spp. causing hypocotyl and root rot on kale Brazil comprises a new AG not yet described.     

Characterization of Rhizoctonia solani Associated with Soybean in Brazil

Rhizoctonia solani causes pre- and post-emergence damping-off, root and hypocotyl rot and foliar blight in soybean. Foliar blight has resulted in yield losses of 31–60% in north and northeast Brazil. The aim of this study was to characterize isolates of R. solani associated with soybean in Brazil. Among 73 Rhizoctonia isolates examined, six were binucleate and 67 were multinucleate. The multinucleate iso1ates were characterized according to hyphal anastomosis reaction, mycelial growth rate, thiamine requirement, sclerotia production, and RAPD molecular markers. Four isolates that caused hypocotyl rot belonged to AG-4 and using RAPD analysis they grouped together with the HGI subgroup. Another isolate that caused root and hypocotyl rots was thiamine auxotrophic, grew at 35°C, and belonged to AG-2-2 IIIB. All 62 isolates that caused foliar blight belonged to AG-1 IA. RAPD analysis of R. solani AG-1 IA soybean isolates showed high genetic similarity to a tester strain of AG-1 IA, confirming their classification. The teleomorph of R. solani, Thanatephorus cucumeris was produced in vitro by one AG-1 IA isolate from soybean. The AG-4 and AG-2-2 IIIB isolates caused damping-off and root and hypocotyl rots of soybean seedlings cv. FT-Cristalina, under greenhouse conditions. The AG-2-2 IIIB isolate caused large lesions on the cortex tissue, that was distinct from the symptoms caused by AG-4 isolates. The AG-1 IA isolates caused foliar blight in adult soybean plants cv. Xingu under the greenhouse and also in a detached-leaf assay.     

Ant28 gene for proanthocyanidin synthesis encoding the R2R3 MYB domain protein (Hvmyb10) highly affects grain dormancy in barley

A number of anthocyanin- and proanthocyanidin-free mutants (ant mutants) in barley were induced and selected because of breeding interest to reduce proanthocyanidins, which could cause haze and degrade the quality of beer. Ant loci, known as anthocyanin or proanthocyanidin synthesis genes, are classified into Ant1 to Ant30 through allelism tests. However, only the Ant18 gene has been molecularly shown to encode dihydroflavonol 4-reductase (DFR), which is involved in both anthocyanin and proanthocyanidin synthesis. In this study, an R2R3 MYB gene of barley was isolated by PCR and named Hvmyb10 due to its similarity to Tamyb10 of wheat, which is a candidate for the R-1 gene grain color regulator. The predicted amino acid sequences of Hvmyb10 showed high similarity not only to Tamyb10 but also to TT2, the proanthocyanidin regulator of Arabidopsis. Non-synonymous nucleotide substitutions in the Hvmyb10 gene were found in all six ant28 mutants tested. Mapping showed that a polymorphism in Hvmyb10 perfectly cosegregated with the ant 28 phenotype on the distal region of the long arm of chromosome 3H. These results demonstrate that ant28 encodes Hvmyb10, the R2R3 MYB domain protein that regulates proanthocyanidin accumulation in developing grains. The reduced grain dormancy of ant28 mutants compared with those of the respective wild types indicates that Hvmyb10 is a key factor in grain dormancy in barley.     

Immuno-magnetic separation and agar layer methods for the isolation of freeze-injured Yersinia enterocolitica O:8 from water

To develop an effective method to isolate an injured pathogenic Yersinia enterocolitica O:8 organism from environmental samples, we compared the isolation of freeze-injured and non-injured Y. enterocolitica O:8 and found that the isolation was more successful when immuno-magnetic separation (IMS) with anti-Y. enterocolitica O:8 antibody was used. Plating onto cefsulodin-irgasan-novobiocin (CIN) agar and Virulent Yersinia enterocolitica (VYE) agar by means of the agar layer method was found to be effective in isolating the injured cells. The alkali treatment which is generally used for selective detection of Yersinia organism failed to isolate freeze-injured pathogenic Y. enterocolitica O:8 cells. Recovery methods without using the alkali treatment were superior for detecting freeze-injured Y. enterocolitica O:8. Our results demonstrate that the IMS and the agar layer methods should be used to isolate injured pathogenic Yersinia organisms from environmental samples such as water.     

Effect of bovine lactoferrin feeding on lipopolysaccharide-induced metabolic and hormonal disturbances in preruminant calves

One of the biological functions of bovine lactoferrin (LF) is modulation of the host defense system, including cytokine production and immune response. The aim of the present study was to investigate the effect of oral administration of LF in calves on lipopolysaccharide (LPS)‐induced metabolic and hormonal changes in inflammatory response. Thirty Holstein calves at 4 day of age were given one of three oral doses of LF (0, 1, 3 g/day) for 10 days (?10 day to ?1 day). They were injected i.v. with LPS (50 ng/kg bodyweight) the day (day 0) after the end of LF treatment. Plasma samples were obtained on ?10, 0 day (immediately before LPS injection), and at 2, 6, 12, 24, 48, 72, and 96 h after LPS injection. Plasma tumor necrosis factor‐α concentrations at 2 h after LPS treatment were lower (P < 0.05) in LF 1 g/day‐fed claves compared with LF 0 g/day (control) calves. On day 0 there were no significant group differences in plasma LF concentration. Plasma concentration of haptoglobin in control calves was elevated by LPS injection. In LF groups, plasma haptoglobin concentrations slightly increased after LPS injection, but those levels at 6–24 h were lower (P < 0.05) than in the control group. The LF treatment inhibited (P < 0.05) the reduction of plasma ferrin concentration in calves following LPS challenge. The concentration of plasma aspartate aminotransferase in calves treated with LF was lower (P < 0.05) than in control calves at 24–96 h after LPS treatment. The concentration of plasma insulin‐like growth factor‐1 (IGF‐1) in all groups was decreased by LPS treatment, while in the LF groups the IGF‐1 level was higher (P < 0.05) than in the control group. Plasma adrenocorticotropic hormone and insulin concentrations in LF groups were lower (P < 0.05) than in control calves at 2 h after LPS injection. These data suggest that LF has a substantial anti‐inflammatory effect on the modulation of the host defense system in preruminant calves.     

Fate of walnut seeds,Juglans airanthifolia,hoarded by Japanese squirrels,Sciurus lis

Walnut seeds (Juglans ailanthiforia Carr.) hoarded by Japanese squirrels (Sciurus lis Temminck) were tracked using miniature radio transmitters. A total of 100 seeds with transmitters were provided on the feeding stands in the walnut forest from September to December, 1992. Squirrels ate 35 seeds without hoarding, and 60 were scatter-hoarded. Five seeds were eaten by wood mice (Apodemus speciosus Temminck). The distance from the feeding stand to cache sites ranged from 1 to 62 m with the mean 18 m. Of 60 hoarded seeds, 38 were retrieved by squirrels later, but 15 were stolen by wood mice. After all, 7 seeds were not retrieved until May 1993.