Production of a desirable Brassica oleracea CMS line using an alloplasmic B. rapa CMS line carrying Diplotaxis erucoides cytoplasm as a bridge plant

In Brassica oleracea, production of F₁ hybrid seeds mainly makes use of the improved Ogura cytoplasmic male sterile (CMS) line. However, reliance on one particular line is a risk, and it would be advantageous to develop other CMS lines. In this study, we transferred Diplotaxis erucoides cytoplasm to B. oleracea cultivars using an alloplasmic B. rapaCMS line as a bridge plant to avoid incompatibility between donor and recipient plants. The new B. oleraceaCMS lines, which were derived by four generations of backcrossing, had small rudimentary anthers with no pollen grain and showed complete male sterility. There was no functional defect in other floral organs, and the ability to receive normal pollen did not appear to be impaired. Moreover, the B. oleraceaCMS lines carrying D. erucoides cytoplasm had larger leaf areas and a normal plastochron. As a consequence, the B. oleraceaCMS lines carrying D. erucoides cytoplasm have the potential to be valuable alternatives for use in commercial B. oleracea hybrid seed production.     

In vivo growth and genomic characterization of rickettsia‐like organisms isolated from farmed Chinook salmon (Oncorhynchus tshawytscha) in New Zealand

A rickettsia‐like organism, designated NZ‐RLO2, was isolated from Chinook salmon (Oncorhynchus tshawytscha) farmed in the South Island, New Zealand. In vivo growth showed NZ‐RLO2 was able to grow in CHSE‐214, EPC, BHK‐21, C6/36 and Sf21 cell lines, while Piscirickettsia salmonis LF‐89^T grew in all but BHK‐21 and Sf21. NZ‐RLO2 grew optimally in EPC at 15°C, CHSE‐214 and EPC at 18°C. The growth of LF‐89 ^T was optimal at 15°C, 18°C and 22°C in CHSE‐24, but appeared less efficient in EPC cells at all temperatures. Pan‐genome comparison of predicted proteomes shows that available Chilean strains of P. salmonis grouped into two clusters (p‐value = 94%). NZ‐RLO2 was genetically different from previously described NZ‐RLO1, and both strains grouped separately from the Chilean strains in one of the two clusters (p‐value = 88%), but were closely related to each other. TaqMan and Sybr Green real‐time PCR targeting RNA polymerase (rpoB) and DNA primase (dnaG), respectively, were developed to detect NZ‐RLO2. This study indicates that the New Zealand strains showed a closer genetic relationship to one of the Chilean P. salmonis clusters; however, more Piscirickettsia genomes from wider geographical regions and diverse hosts are needed to better understand the classification within this genus.     

Liquid Chromatography-Mass Spectrometry-Based Rapid Secondary-Metabolite Profiling of Marine Pseudoalteromonas sp. M2

The ocean is a rich resource of flora, fauna, and food. A wild-type bacterial strain showing confluent growth on marine agar with antibacterial activity was isolated from marine water, identified using 16S rDNA sequence analysis as Pseudoalteromonas sp., and designated as strain M2. This strain was found to produce various secondary metabolites including quinolone alkaloids. Using high-resolution mass spectrometry (MS) and nuclear magnetic resonance (NMR) analysis, we identified nine secondary metabolites of 4-hydroxy-2-alkylquinoline (pseudane-III, IV, V, VI, VII, VIII, IX, X, and XI). Additionally, this strain produced two novel, closely related compounds, 2-isopentylqunoline-4-one and 2-(2,3-dimetylbutyl)qunoline-4-(1H)-one, which have not been previously reported from marine bacteria. From the metabolites produced by Pseudoalteromonas sp. M2, 2-(2,3-dimethylbutyl)quinolin-4-one, pseudane-VI, and pseudane-VII inhibited melanin synthesis in Melan-A cells by 23.0%, 28.2%, and 42.7%, respectively, wherein pseudane-VII showed the highest inhibition at 8 µg/mL. The results of this study suggest that liquid chromatography (LC)-MS/MS-based metabolite screening effectively improves the efficiency of novel metabolite discovery. Additionally, these compounds are promising candidates for further bioactivity development.     

Effect of Phlorofucofuroeckol A and Dieckol Extracted from Ecklonia cava on Noise-induced Hearing Loss in a Mouse Model

One of the well-known causes of hearing loss is noise. Approximately 31.1% of Americans between the ages of 20 and 69 years (61.1 million people) have high-frequency hearing loss associated with noise exposure. In addition, recurrent noise exposure can accelerate age-related hearing loss. Phlorofucofuroeckol A (PFF-A) and dieckol, polyphenols extracted from the brown alga Ecklonia cava, are potent antioxidant agents. In this study, we investigated the effect of PFF-A and dieckol on the consequences of noise exposure in mice. In 1,1-diphenyl-2-picrylhydrazyl assay, dieckol and PFF-A both showed significant radical-scavenging activity. The mice were exposed to 115 dB SPL of noise one single time for 2 h. Auditory brainstem response(ABR) threshold shifts 4 h after 4 kHz noise exposure in mice that received dieckol were significantly lower than those in the saline with noise group. The high-PFF-A group showed a lower threshold shift at click and 16 kHz 1 day after noise exposure than the control group. The high-PFF-A group also showed higher hair cell survival than in the control at 3 days after exposure in the apical turn. These results suggest that noise-induced hair cell damage in cochlear and the ABR threshold shift can be alleviated by dieckol and PFF-A in the mouse. Derivatives of these compounds may be applied to individuals who are inevitably exposed to noise, contributing to the prevention of noise-induced hearing loss with a low probability of adverse effects.     

Intra-arterial delivery of triolein emulsion increases vascular permeability in skeletal muscles of rabbits

Background

To test the hypothesis that triolein emulsion will increase vascular permeability of skeletal muscle.

Methods

Triolein emulsion was infused into the superficial femoral artery in rabbits (triolein group, n = 12). As a control, saline was infused (saline group, n = 18). Pre- and post-contrast T1-weighted MR images were obtained two hours after infusion. The MR images were qualitatively and quantitatively evaluated by assessing the contrast enhancement of the ipsilateral muscles. Histologic examination was performed in all rabbits.

Results

The ipsilateral muscles of the rabbits in the triolein group showed contrast enhancement, as opposed to in the ipsilateral muscles of the rabbits in the saline group. The contrast enhancement of the lesions was statistically significant (p < 0.001). Histologic findings showed that most examination areas of the triolein and saline groups had a normal appearance.

Conclusion

Rabbit thigh muscle revealed significantly increased vascular permeability with triolein emulsion; this was clearly demonstrated on the postcontrast MR images.     

Influence of equalizing the gross composition of milk replacer to that of whole milk on the performance of Holstein calves

This study compared the performance of female Holstein calves fed either whole milk (WM) or milk replacer (MR) having similar gross composition to WM. Calves (n = 20) were separated from their mothers within 2 h of birth, weighed, moved into individual pens, and fed colostrum (10% of their BW over 2 feedings daily) for the first 3 d. Calves were alternately assigned to either WM (n = 10) or MR (n = 10) and were fed using mobile plastic bottles. At each feeding, diluted MR was prepared in buckets by mixing 0.143 kg of MR powder (as-is basis) in 1 L of lukewarm water. Calves on both treatments were fed (1.8 L/feeding) for 4 times daily for the first 25 d of age. Feeding frequency was reduced to 3 times daily for next 5 d, then to 2 times daily for next 14 d, and then to once daily during the last 5 d of the preweaning period. Feed intake, growth, and health variables were monitored until calves were 70 d of age. Mean daily consumption of WM and MR was similar in both treatment groups (P = 0.74). Initial BW of calves fed either WM or MR was similar (42.1 +/- 3.3 vs. 41.9 +/- 2.9 kg, respectively; P = 0.83). Calves fed WM or MR daily consumed similar amounts of calf starter (1,019 vs. 1,056 g, respectively; P = 0.32), hay (121.5 vs. 126.3 g, respectively; P = 0.30), and water (5.2 vs. 5.0 kg, respectively; P = 0.54). At weaning (d 49) and postweaning (d 70), BW was greater (P = 0.03 and P = 0.02, respectively) in calves fed WM than in those fed MR. Mean DMI from both solid and liquid feeds was not affected by the treatments during the preweaning (P = 0.21) and postweaning (P = 0.16) periods. Body weight gain efficiency during preweaning and overall was improved (P = 0.001 and P = 0.002, respectively) in calves fed WM than in those fed MR. Health (days scoured, rectal temperature, respiratory score, and general appearance score) and serum chemistry variables (glucose, total protein, urea N, nonesterified fatty acids, and creatinine) in calves were not affected (P >or= 0.12 and P >or= 0.12, respectively) by the treatments. Even though gross composition of the MR and WM was similar, growth was greater in calves fed WM. Calves fed WM consumed similar amounts of DM and were heavier than those fed MR, probably because of better bioavailability (digestion and assimilation) of nutrients and availability of some unknown growth factors from WM.     

An outbreak of fatal hemorrhagic pneumonia caused by Streptococcus equi subsp. zooepidemicus in shelter dogs

An outbreak of fatal hemorrhagic pneumonia with 70~90% morbidity and 50% mortality occurred in an animal shelter in Yangju, Gyeonggi Province, Korea. Clinically, the affected dogs showed severe respiratory distress within 48 h after arriving in the shelter. The dead were found mainly with nasal bleeding and hematemesis. At necropsy, hemothorax and hemorrhagic pneumonia along with severe pulmonary consolidation was observed, though histopathological analysis showed mainly hemorrhagic bronchopneumonia. Lymphoid depletion was inconsistently seen in the spleen, tonsil and bronchial lymph node. Gram-positive colonies were shown in blood vessels or parenchyma of cerebrum, lung, liver, spleen, and kidney. Also, Streptococcus (S.) equi subsp. zooepidemicus was isolated from the various organs in which the bacterium was microscopically and histologically detected. In addition, approximately 0.9 Kb specific amplicon, antiphagocytic factor H binding protein, was amplified in the bacterial isolates. In this study, we reported an outbreak of canine hemorrhagic bronchopneumonia caused by S. equi subsp. zooepidemicus in an animal shelter in Yangju, Korea.     

Evaluation of a competitive ELISA for antibody detection against avian influenza virus

Active serologic surveillance is necessary to control the spread of the avian influenza virus (AIV). In this study, we evaluated a commercially-available cELISA in terms of its ability to detect AIV antibodies in the sera of 3,358 animals from twelve species. cELISA detected antibodies against reference H1- through H15-subtype AIV strains without cross reactivity. Furthermore, the cELISA was able to detect antibodies produced following a challenge of the AIV H9N2 subtype in chickens, or following vaccination of the AIV H9 or H5 subtypes in chickens, ducks and geese. Next, we tested the sensitivity and specificity of the cELISA with sera from twelve different animal species, and compared these results with those obtained by the hemagglutination-inhibition (HI) test, the "gold standard" in AIV sera surveillance, a second commercially-available cELISA (IZS ELISA), or the agar gel precipitation (AGP) test. Compared with the HI test, the sensitivities and specificities of cELISA were 95% and 96% in chicken, 86% and 88% in duck, 97% and 100% in turkey, 100% and 87% in goose, and 91% and 97% in swine, respectively. The sensitivities and specificities of the cELISA in this study were higher than those of IZS ELISA for the duck, turkey, goose, and grey partridge sera samples. The results of AGP test against duck and turkey sera also showed significant correlation with the results of cELISA (R-value >0.9). In terms of flock sensitivity, the cELISA correlated better with the HI test than with commercially-available indirect ELISAs, with 100% flock sensitivity.     

Ozone impairs autumnal resorption of nitrogen from birch (Betula pendula) leaves, causing an increase in whole-tree nitrogen loss through litter fall

Saplings of one half-sib family of birch, Betula pendula Roth, were exposed to three ozone concentrations (non-filtered air (NF); non-filtered air + 10-20 nmol O(3) mol(-1) (NF+); non-filtered air + 40-60 nmol O(3) mol(-1) (NF++)) in open-top chambers during two growing seasons from 1997 to 1998. Shed leaves were collected regularly during both growing seasons and, in 1998, the dry mass (DM) and nitrogen (N) concentrations ([N]) of the shed leaves were measured to quantify the total amount of N lost through litter fall. Dry mass and [N] were also determined in mid-August for attached, mature and non-senescent leaves, in order to estimate autumnal leaf N resorption efficiency and proportional leaf DM decrease. Net photosynthetic capacity was measured during August and September 1998, in a population of leaves that emerged in mid-July. Photosynthesis declined with increasing leaf age in the NF++ treatment, whereas it remained high throughout the measurement period in the NF and NF+ treatments. In both years, leaves abscised prematurely in the NF++ treatment, whereas this effect was only significant in 1998 in the NF+ treatment. There was a strong linear relationship between proportional leaf shedding and daylight ozone exposure above a threshold of 40 nmol mol(-1) (daylight AOT40) during the growing season. The resorption of N was significantly impaired by ozone, and the smaller autumnal decrease in leaf DM in elevated ozone concentrations suggested that the bulk resorption of leaf DM was also inhibited. Nitrogen resorption efficiencies were 81, 73 and 63% and leaf mass decreases were 45, 36 and 30% in the NF, NF+ and NF++ treatments, respectively. Compared with the NF treatment, total N loss through litter fall was increased by 16 and 122% in the NF+ and NF++ treatments, respectively. We conclude that ozone impaired N resorption from birch leaves before abscission, causing a substantial increase in whole-tree N loss through litter fall.