Identification of Rfd1, a novel restorer-of-fertility locus for cytoplasmic male-sterility caused by DCGMS cytoplasm and development of simple PCR markers linked to the Rfd1 locus in radish (Raphanus sativus L.)

Previously, novel cytoplasmic male-sterility (CMS) caused by DCGMS cytoplasm was discovered in radish (Raphanus sativus L.) introduced from Uzbekistan. We performed extensive progeny tests and identified two fertility restorer lines (‘R171’ and ‘R121’) for this new CMS. Two F₁ hybrid populations were self-pollinated and backcrossed to produce F₂ and BC populations. Inheritance patterns of male-sterility in segregating populations varied depending on paternal lines. Segregation of male-sterility in F₂ populations originating from the cross between MS19 and R121 showed that a single locus was involved in fertility restoration. However, populations originating from the cross between MS15 and R171 showed the involvement of more than one restorer-of-fertility genes. The single fertility restorer locus identified in the cross between MS19 and R121 was designated Rfd1 locus. Bulked segregant analysis was performed using RAPD and AFLP, which identified one marker each. Both RAPD and AFLP markers were converted into simple PCR-based co-dominant markers after their isolated flanking sequences were analyzed. Indels 773-bp and 67-bp in length were identified between two Rfd1 allele-linked flanking sequences of the RAPD and AFLP fragments, respectively, then utilized to develop simple PCR markers. In addition, we prove that the newly identified Rfd1 locus is independent of the Rfo locus, another radish fertility restorer for CMS caused by Ogura cytoplasm.     

Identification of a cluster of oligonucleotide repeat sequences and its practical implication in melon (<Emphasis Type="Italic">Cucumis melo</Emphasis> L.) breeding

A cluster of oligonucleotide repeat sequences was identified that is linked to the melon (Cucumis melo L.) andromonoecious (a) locus, which controls andromonoecy. Six different repeat units, ranging from 11 to 49 base pairs, were clustered within a 0.6-kb intergenic region. The number of repeat units varied from two to six. After sequence analysis of diverse melon germplasms, four haplotypes of this cluster were identified. Length variations among the four haplotypes resulted from insertion or deletion of repeat sequences. Particularly, a tandem array of six repeats of 21 nucleotides was a hotspot for insertion/deletion mutations. A simple PCR-based marker was developed to identify haplotypes of this cluster based on the length polymorphism. In practice, this marker was successfully used in genetic purity tests of melon F₁ hybrid cultivars. Four self-pollinated contaminants, which were confirmed by phenotypic examination in grow-out tests, were easily discriminated from 99 F₁ hybrid individuals. In addition, the genetic distance between this marker and the andromonoecious (a) locus was calculated as 7.9 cM, after analyzing melon F₂ populations originating from a cross between monoecious and andromonoecious parental lines. Therefore, this marker will be useful as a recombinant selection marker in marker-assisted backcrossing of monoecy in melon breeding programs.     

Multidetector row computed tomography evaluation of the micropig kidney as a potential renal donor

Multidetector row computed tomography (MDCT) provides anatomical information about the kidney and other internal organs. Presently, the suitability of 64-channel MDCT to assess the kidney of healthy micropigs was evaluated. Morphological evaluations of the kidney and the major renal vessels of six healthy micropigs were carried out using MDCT, recording kidney volume and the diameter and length of renal arteries and veins. The mean diameters and lengths of the renal artery were 0.44 ± 0.05 and 4.51 ± 0.55 cm on the right side and 0.46 ± 0.06 and 3.36 ± 0.27 cm on the left side, respectively. The mean diameters and lengths of the renal vein were 1.44 ± 0.52 and 4.22 ± 1.29 cm on the right side and 1.38 ± 0.17 and 5.15 ± 0.87 cm on the left side, respectively. The mean volume of the right kidney was 79.3 ± 14.5 mL and of the left kidney was 78.0 ± 13.9 mL. The data presented in this study suggest that the MDCT offers a noninvasive, rapid, and accurate method for the evaluation of the renal anatomy in living kidney donors. It also provides sufficient information about extra-renal anatomy important for donor surgery and determination of organ suitability.     

Ingestion of food pellets containing Escherichia coli overexpressing the heat‐shock protein DnaK protects Penaeus vannamei (Boone) against Vibrio harveyi (Baumann) infection

Feeding aquatic animals with bacterial encapsulated heat‐shock proteins (Hsps) is potentially a new method to combat vibriosis, an important disease affecting aquatic animals used in aquaculture. Food pellets comprised of shrimp and containing Escherichia coli overexpressing either DnaK‐DnaJ‐GrpE, the prokaryotic equivalents of Hsp70‐Hsp40‐Hsp20, or only DnaK were fed to juveniles of the white leg shrimp Penaeus vannamei, and protection against pathogenic Vibrio harveyi was determined. Maintaining pellets at different temperatures for varying lengths of time reduced the number of live adhering E. coli, as did contact with sea water, demonstrating that storage and immersion adversely affected bacterial survival and attachment to pellets. Feeding P. vannamei with E. coli did not compromise their survival, indicating that the bacteria were not pathogenic to shrimp. Feeding P. vannamei with pellets containing bacteria overproducing DnaK (approximately 60 cells g^?1 pellets) boosted P. vannamei survival twofold against V. harveyi, suggesting that DnaK plays a role in Vibrio tolerance. Pellets containing DnaK were effective in providing protection to P. vannamei for up to 2 weeks before loss of viability and that DnaK encapsulated by these bacteria enhanced shrimp resistance against Vibrio infection.     

Effects of dietary substitution of fishmeal with the combined dry microalgae,Nannochloropsis oceanica (NO) biomass residue and casein on growth and body composition of juvenile abalone (Haliotis discus)

The effects of dietary substitution of fishmeal with the combined dry Nannochloropsis oceanica (NO) biomass residue and casein on growth performance and carcass composition of juvenile abalone was determined. One thousand two hundred and sixty juvenile abalone were randomly distributed into the 18 L plastic rectangular containers. Five experimental diets in triplicate were prepared. The 28% fishmeal was included into the control (Con) diet. The 25%, 50%, 75% and 100% fishmeal were substituted with the combination of graded levels of dry NO biomass residue and casein, referred to as the NO25, NO50, NO75 and NO100 diets respectively. Finally, the salted sea tangle (ST), Laminaria japonica was prepared to compare the effect of the experimental diets on performance of abalone. The essential amino acids, such as isoleucine, leucine, phenylalanine, threonine and valine tended to increase with dietary substitution of fishmeal with NO biomass residue in the experimental diets. Arginine tended to decrease with dietary substitution of fishmeal with NO biomass residue. Survival of abalone fed the experimental diets was higher than that of abalone fed the ST diet for 16 weeks. Weight gain and specific growth rate (SGR) of abalone fed the NO100 diet were higher than those of abalone fed the all other diets. Weight gain and SGR of abalone linearly increased with dietary substation of fishmeal with NO. Shell length of abalone tended to increase with dietary substitution of fishmeal with the combined dry NO biomass residue and casein. The chemical composition of the soft body of abalone was different among treatments except for moisture content. In conclusion, fishmeal in the diets for abalone could be completely replaced with the combined dry NO biomass residue and casein when the 28% fishmeal was included.     

Revitalizing Mountain Ginseng Cultivation in North Jeolla Province,South Korea

A study was made of measures to revitalize mountain ginseng cultivation in the eastern mountainous area of Muju, Jinan and Jangsu municipalities of North Jeolla province, South Korea. The targeted area is typical forested land and is utilized for the cultivation of ginseng. Access to seeds and seedlings for mountain ginseng cultivation is relatively easy in North Jeolla province, and the species is cultivated widely (on 63 % of the area of the province and by 74 % of farmers). This study investigated the current state of mountain ginseng cultivation and business management, including production costs and subsidy, sales and distribution system, and other business management concerns. A survey was conducted on 86 mountain ginseng-cultivating farming households living in the targeted area in 2013. It is concluded that to achieve greater production an aggressive government policy promoting the renting out of suitable sites for mountain ginseng cultivation in national and public production forestland to individual farms and farming cooperatives is necessary. Further research on increasing the survival rate of cultivated mountain ginseng (CMG) until harvesting and preventing damage by wild animals is also deemed necessary. More effective use of related municipal budget subsidy needs to be supported selectively to the enterprising farmers who aggressively pursue mountain ginseng cultivation and business management. Opening CMG retail stores to realize sustainable income growth and developing various processed products will increase farm income. The government needs to introduce policies to sustain consumer trust in CMG through initiatives such as establishing a CMG quality management system for growers, distributing government-approved seeds, and implementing a government-based CMG purchase system.     

Characterization of three chitosanase isozymes isolated from a commercial crude porcine pepsin preparation

Three chitosanases designated PSC-I, PSC-II, and PSC-III were purified from commercial pepsin preparation by sequentially applying pepstatin A-agarose affinity chromotography, DEAE-Sephacel ion-exchange chromatography, Mono Q column chromatography, and Mono P chromatofocusing. With respect to chitosan hydrolysis, the optimal pHs were 5.0, 5.0, and 4.0 for PSC-I, PSC-II, and PSC-III, respectively; optimal temperatures were 40, 40, and 30 degrees C; and the Km's were 5.2, 4.0, and 5.6 mg/mL. The molecular masses of the three isozymes were approximately 40 kDa, as estimated by both gel filtration and SDS-PAGE, and the isoelectric points were 4.9, 4.6, and 4.4, respectively, as estimated by isoelectrofocusing electrophoresis. All three chitosanase isozymes showed activity toward chitosan polymer and N,N",N' "-triacetylchitotriose oligomer. Most effectively hydrolyzed were chitosan polymers that were 68-88% deacetylated.     

Over representation of Burmese cats with diabetes mellitus

Objective To determine if Burmese cats in Queensland have an increased risk of diabetes mellitus.
Design A retrospective study of diabetic and nondiabetic cats that had blood submitted to a veterinary clinical laboratory over a 22 month study period.
Sample population 4402 cats
Procedure Cats were considered diabetic if blood glucose concentration was > 11 mmol/L and fructosamine was > 406 μmol/L or hydroxybutyrate was >1 mmol/L. Cats were grouped into Burmese and non-Burmese. Adjusted odds ratios of diabetes were calculated for breed, gender and age group amongst cats with blood glucose > 11 mmol/L.
Results Burmese cats comprised 20% of 45 diabetic cats of known breed, which was higher (P < 0.001) than among the normoglycemic reference population of 2203 cats (7% Burmese). There were more females among the diabetic Burmese (62%), but this did not differ (P > 0.05) from the Burmese reference population (45% females). In contrast, males seemed to predominate among diabetic non-Burmese (63%), although this also did not differ (P > 0.05) from the reference population (55%) or from diabetic Burmese (38% males). The majority (90%) of diabetic cats were older than 6 years, irrespective of breed (median age 12 years, interquartile range 10 to 13 years). This was higher (χ²= 8.13, P < 0.005) than among the normoglycaemic reference population, where 69% were older than 6 years.
Conclusions Burmese cats were significantly over represented among cats with diabetes mellitus. Irrespective of breed, the risk of diabetes in the study population increased with age.     

Toxoplasma gondii: level of carriage in sheep of Marrakech region (Mnabha)

Toxoplasma gondii is an ubiquitous parasite with a prevalence variable from country to country. In Morocco very few studies were devoted to this prevalence. To fill this gap we were interested to study the epidemiology of this parasite and to know the level of carriage by the different vectors which are the sources of contamination in humans. The study was done by directly detecting the cysts in the cerebral tissue of the 50 sheep killed and destined for consumption. The results of this preliminary study show that 30% of the cases carry the cysts of T. gondii. To confirm this result and verify the virulence, cerebral specimens were inoculated into mice. These findings are encouraging to complete this study with serological tests and to look for the parasite in cows and goats of this region.