Reproductive losses associated with porcine circovirus type 2 in a Japanese herd of seronegative sows

From April to July 2009, there was a high rate of reproductive losses in a 30-sow, farrow-to-finish swine herd in Yamagata Prefecture, Japan. Histopathological examinations of heart tissue from stillborn and preweaning piglets showed nonsuppurative, necrotizing lesions. Immunohistochemical staining and polymerase chain reaction analysis of myocardial lesions revealed the presence of porcine circovirus type 2 (PCV2) antigens and DNA in these tissues. Indirect immunofluorescence also showed that the PCV2 antibody positive rate in the sows was higher in May 2009 than in December 2008. The results of this study suggest that PCV2 spread to this farm and caused a high rate of reproductive losses.     

New Cyclic Lipopeptides of the Iturin Class Produced by Saltern-Derived Bacillus sp. KCB14S006

Salterns, one of the most extreme natural hypersaline environments, are a rich source of halophilic and halotolerant microorganisms, but they remain largely underexplored ecological niches in the discovery of bioactive secondary metabolites. In continued efforts to investigate the metabolic potential of microbial populations from chemically underexplored sites, three new lipopeptides named iturin F₁, iturin F₂ and iturin A₉ (1–3), along with iturin A₈ (4), were isolated from Bacillus sp. KCB14S006 derived from a saltern. The structures of the isolated compounds were established by 1D-, 2D-NMR and HR-ESIMS, and their absolute configurations were determined by applying advanced Marfey’s method and CD spectroscopy. All isolates exhibited significant antifungal activities against various pathogenic fungi and moderate cytotoxic activities toward HeLa and src^ts-NRK cell lines. Moreover, in an in vitro enzymatic assay, compound 4 showed a significant inhibitory activity against indoleamine 2,3-dioxygenase.     

Optimization of the supplemental essential amino acids to a fish meal-free diet based on fermented soybean meal for rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis>

To minimize the supplemental essential amino acids (EAAs) to a fish meal-free diet for rainbow trout Oncorhynchus mykiss, four types of fish meal-free diets and a control fish meal-based diet were fed to triplicate groups of trout (initial BW, 16 g) for 10 weeks. Two fish meal-free diets based on a fermented and an unfermented soybean meal were unsupplemented with EAAs (diet FSBM and SBM), and two fish meal-free diets based on the fermented soybean meal were supplemented with a combination of lysine and methionine (diet F + ML) and all EAAs (diet F + All), the EAA contents of which were lower than those of the control diet (diet FM). Although physiological conditions such as the biliary bile acid status and morphological features of fish fed diet FSBM were improved and similar to those of fish fed diet FM, the growth performance was not significantly different (P > 0.05) from fish fed diet SBM. The growth performances and whole body protein contents of fish fed F + ML and F + All diets were similarly improved compared to fish fed diet FSBM. These findings indicate that supplementation of lysine and methionine to a fermented soybean meal-based fish meal-free diet is enough to maximize the amino acid utilization of the diet.     

Central estrogen action sites involved in prepubertal restraint of pulsatile luteinizing hormone release in female rats

The present study aimed to determine estrogen feedback action sites to mediate prepubertal restraint of gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) release in female rats. Wistar-Imamichi strain rats were ovariectomized (OVX) and received a local estradiol-17β (estradiol) or cholesterol microimplant in several brain areas, such as the medial preoptic area (mPOA), paraventricular nucleus, ventromedial nucleus and arcuate nucleus (ARC), at 20 or 35 days of age. Six days after receiving the estradiol microimplant, animals were bled to detect LH pulses at 26 or 41 days of age, representing the pre- or postpubertal period, respectively. Estradiol microimplants in the mPOA or ARC, but not in other brain regions, suppressed LH pulses in prepubertal OVX rats. Apparent LH pulses were found in the postpubertal period in all animals bearing estradiol or cholesterol implants. It is unlikely that pubertal changes in responsiveness to estrogen are due to a change in estrogen receptor (ER) expression, because the number of ERα-immunoreactive cells and mRNA levels of Esr1, Esr2 and Gpr30 in the mPOA and ARC were comparable between the pre- and postpubertal periods. In addition, kisspeptin or GnRH injection overrode estradiol-dependent prepubertal LH suppression, suggesting that estrogen inhibits the kisspeptin-GnRH cascade during the prepubertal period. Thus, estrogen-responsive neurons located in the mPOA and ARC may play key roles in estrogen-dependent prepubertal restraint of GnRH/LH secretion in female rats.     

Detection of Salmonella invA by isothermal and chimeric primer-initiated amplification of nucleic acids (ICAN) in Zambia

The isothermal and chimeric primer-initiated amplification of nucleic acids (ICAN) is a new isothermal DNA amplification method composed of exo Bca DNA polymerase, RNaseH and DNA–RNA chimeric primers. We detected invA of Salmonella from chicken carcasses, egg yolk and cattle fecal samples. Fifty-three of 59 isolates were invA-positive in ICAN-chromatostrip detection. The result was consistent with those obtained by standard PCR. Salmonella invA was detected in 12 of 14 carcass rinses by ICAN, while in 7 of 14 rinses by standard PCR. These results indicate that ICAN is an efficient, sensitive and simple system to detect invA of Salmonella species in developing countries such as Zambia.     

Susceptibility difference between methicillin‐susceptible and methicillin‐resistant Staphylococcus aureus to a bovine myeloid antimicrobial peptide (BMAP‐28)

A bovine myeloid antimicrobial peptide antimicrobial peptide (BMAP‐28) is a member of the cathelicidin family and acts as a component of innate immunity. There are few reports of susceptibility difference of methicillin‐resistant Staphylococcus aureus (MRSA) and susceptible strains (MSSA) against BMAP‐28. This study aims to clarify how a few amino acid substitutions of BMAP‐28 are related to its antimicrobial activity using four analog peptides of BMAP‐28. We also compared cellular fatty acid components of MSSA and MRSA using gas chromatography. We found that a few amino acid substitutions of BMAP‐28 do not change antimicrobial activity. It was also revealed that the percentage of cis‐11‐eicosenoic acid in total detected fatty acids of MRSA was significantly higher than that of MSSA. In addition, the percentage of palmitic acid in total detected fatty acids of MRSA tended to be lower than that of MSSA. Our results will provide new information to deal with the question of differences in bacterial susceptibility against BMAP‐28.     

Translocation and distribution of assimilated carbon in peanut plant

An attempt was made to monitor ¹³C that had been photosynthetically assimilated in the foliage of the main stem and branches of peanut plant, as well as in a single leaf at different positions on a branch.

When the foliage of the main stem or branch was supplied with ¹³CO₂ for 8 h at the vegetative stage, ¹³C assimilated in the branches was detected in the roots and nodules in addition to the foliage immediately after the exposure, whereas when the main stem was supplied with ¹³CO₂, ¹³C was not detected in the roots and nodules immediately after ¹³CO₂ feeding. At the reproductive stage, ¹³C assimilated in the main stem or branch was found in the leaves, stems, fruit (shell, seed coat, and seed), roots, and nodules immediately after assimilation.

Photoassimilates from each leaf of the branch at the reproductive stage were exported to the fruit and leaves that were attached to the same branch. Namely, photoassimilates in the leaves of odd nodes were mainly translocated to the fruits attached to the first node, whereas such photoassimilates from the leaf of even nodes were mainly translocated to the fruit attached to the second node.

When the foliage of a branch had been fed ¹³CO₂ at the vegetative stage, the loss of the assimilated ¹³C by respiration was about 40% of the total assimilated ¹³C within 23 d and about 65% within 93 d after the exposure, and a small amount of photoassimilates was detected in the fruit. On the other hand, at the seed-filling stage, about 35% of the photoassimilates were utilized for seed growth within 10 d after the end of exposure.

These results suggest that in the peanut plant, the carbon source of nodules mainly depends on the branch, and the main stem plays an important role as carbon source for the fruit, that a sink organ for carbon is connected with a specific sources leaf by the vascular bundles, and that most of the carbon sources for the growth of peanut fruit depend on the photoassimilates at the reproductive stage.     

Nested PCR‐RFLP is a high‐speed method to detect fungicide‐resistant Botrytis cinerea at an early growth stage of grapes

BACKGROUND: Grey mould caused by the fungus Botrytis cinerea Pers. ex Fr. is one of the major diseases in grapes. The use of fungicides is a simple strategy to protect grapes against B. cinerea disease. However, phenotypes exhibiting resistance to fungicides have been detected in B. cinerea populations. The variation of fungicide‐resistant B. cinerea isolates renders B. cinerea disease control difficult in grapevine fields. RESULTS: The authors have developed a nested polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) method to detect fungicide‐resistant B. cinerea isolates at an early growth stage of grapes in grapevine fields. The nested PCR‐RFLP method was carried out to detect benzimidazole‐, phenylcarbamate‐ and/or dicarboximide‐resistant B. cinerea isolates from grape berries and leaves at Eichorn–Lorenz growth stage 25 to 29. This method successfully detected fungicide‐resistant B. cinerea isolates at an early growth stage of grapes. In addition, only 8 h was required from tissue sampling to phenotyping of fungicide resistance of the isolates. CONCLUSION: It is proposed that the early diagnosis of fungicide‐resistant B. cinerea isolates would contribute to further improvement of integrated pest management against B. cinerea in grapevine fields, and that the nested PCR‐RFLP method is a high‐speed, sensitive and reliable tool for this purpose. Copyright © 2008 Society of Chemical Industry     

Evaluation of validity and limitations of the soybean canopy height‐to‐row spacing ratio as an onsite index to control weeds using diverse soybean accessions

A canopy height‐to‐row spacing ratio (CHRSR) of 1.0 is expected to be a simple and practical onsite index for soybean growers to determine when further weed control is no longer necessary. This is decided when relative photosynthetic photon flux density (PPFD) reaches 50% under the soybean canopy. To evaluate the validity of this index, a 2‐year field experiment was conducted to compare days after sowing (DAS) to reach a relative PPFD of 50% (defined as PPFD50) and DAS to reach a CHRSR of 1.0 (defined as CHRSR1.0) in 190 accessions with different canopy architectures from a soybean core collection. A total of 104 accessions, for which final relative PPFD reached <10% in 2014, were examined for the differences between CHRSR1.0 and PPFD50. The mean and median values of the difference in 2014 were 0.02 and 2.2, respectively. Similarly, the mean and median values in 2015 were 2.8 and 2.6, respectively. Thus, CHRSR1.0 tended to be achieved slightly later than PPFD50, so a consideration was made that there would be a negligible risk of failure if growers used CHRSR1.0 to determine the termination time for weed control in the most of accessions. Although there were some accessions with a standing type of morphological characteristics or with a smaller or narrower leaf, showing negative differences between CHRSR1.0 and PPFD50 larger than 1 week, a CHRSR of 1.0 could be used to estimate PPFD50 in most of accessions without risk of failure to control weeds.     

Development of a multiplex allele‐specific primer PCR assay for simultaneous detection of QoI and CAA fungicide resistance alleles in Plasmopara viticola populations

BACKGROUND: DNA‐based diagnosis has become a common tool for the evaluation of fungicide resistance in obligate phytopathogenic fungus Plasmopara viticola. RESULTS: A multiplex allele‐specific primer PCR assay has been developed for the rapid detection of fungicide resistance in P. viticola populations. With this assay, a glycine‐to‐alanine substitution at codon 143 of the P. viticola cytochrome b gene, which conferred QoI fungicide resistance, and a glycine‐to‐serine substitution at codon 1105 of the P. viticola cellulose synthase gene PvCesA3, which conferred CAA fungicide resistance, were detected simultaneously. CONCLUSION: It is suggested that the present assay is a reliable tool for the rapid and simultaneous detection of QoI and CAA fungicide resistance alleles in P. viticola populations. The assay required only 2 h from the sampling of symptoms to the detection of resistance alleles to both fungicides. Copyright © 2012 Society of Chemical Industry