Analysis of cross-reactivity of five new chicken monoclonal antibodies which recognize the apical complex of Eimeria using confocal laser immunofluorescence assay

For Apicomplexa (members) the host cell invasion is realized with the help of the organelles located at the apical tip of parasites. In this research paper the characterization of five chicken monoclonal antibodies (mabs) produced against Eimeria acervulina sporozoites is described. All mabs reacted with molecules belonging to the apical complex of chicken Eimeria sporozoites. On immunofluorescence assay (IFA) one mab, 8E-1, recognized an apical tip molecule present on all chicken Eimeria sporozoites, two mabs (8D-2 and HE-4) recognized an antigen present on the apical tip of the same two Eimeria species (E. acervulina and E. brunetti), another mab (5D-11) recognized an antigen present on the apical tip of other two species (E. acervulina and E. maxima) while one mab (8C-3) identified antigens present on the sporozoites and sporocysts wall of only E. acervulina. Besides the apical tip antigens, two mabs (HE-4 and 8D-2) recognized some proteins located in the anterior half of the sporozoites. Collectively, these mabs proved that the apical complex of chicken Eimeria sporozoites share one or more antigens that are expected to play a role in host cell recognition and invasion.     

Epidemiological studies on the expectation of life for cats computed from animal cemetery records

Based on the Chiang's method, the life table for cats was constructed from the death data of 3936 cats. They died in the Kanto area and were buried in an animal cemetery in Tokyo from June 1981 through May 1982. This life table seems to be the first one for domestic pet cats. The expectation of life for cats was 4.2 years at age 0, 5.0 years at age 1, 5.4 years at age 4, 5.3 years at age 5, 3.5 years at age 10, and 2.2 years at age 15. The maximum age at death was 22 years. From age 0 to age 5, the probability of dying for cats was higher than that for dogs, but over 6 years of age it seemed that Gompertz's equation was applicable to this life table for cats. From these results, if the probability of dying for cats at early ages decreases, the fundamental pattern of dying curve for cats seems to be a similar figure of dogs. The life table was constructed for different breeds and localities. Comparing the expectation of life at age 1 (e1) of the two populations divided by breeds or localities, there was significant difference in the e1 among different localities but not among different breeds. These facts suggest the existence of some factors which may influence the life span of cats among different localities.     

Molecular Characterization and Diagnosis of QoI Resistance in Cucumber and Eggplant Fungal Pathogens

ABSTRACT The molecular mechanism of QoI fungicide resistance was studied using isolates of cucumber Corynespora leaf spot fungus (Corynespora cassiicola) and the eggplant leaf mold (Mycovellosiella nattrassii). In both pathogens, a mutation at position 143 from glycine to alanine (G143A) was detected in the cytochrome b gene that encodes for the fungicide-targeted protein. Moreover, the nucleotide sequence at amino acid position 143 was converted from GGT or GGA in sensitive (wild-type) to GCT or GCA in resistant (mutant-type) isolates. The methods of polymerase chain reaction restriction fragment length polymorphism commonly used for QoI resistance monitoring were employed successfully, leading to the amplified gene fragment from resistant isolates being cut with the restriction enzyme ItaI. However, heteroplasmy (the coexistence of wild-type and mutated alleles) was found when the resistant isolates of C. cassiicola, M. nattrassii, and Colletotrichum gloeosporioides (strawberry anthracnose fungus) were subcultured in the presence or absence of QoI fungicides. QoI resistance of cucumber powdery and downy mildew isolates persisted for a few years following the removal of the selection pressure imposed by the fungicide under both laboratory and commercial greenhouse conditions. The proportion of mutated sequences in cytochrome b gene decreased over time in the pathogen population. The protective efficacy of the full dose of azoxystrobin decreased when the populations of powdery and downy mildews contained resistant isolates at 10%. Using FMBIO, a fluorescence bio-imaging analyzer, the mutant allele from the QoI-resistant isolates could be detected at the level of 1%, whereas the detection sensitivity of ethidium-bromide-stained gels was approximately 10 times lower.     

Variation in the response of melon genotypes to Fusarium oxysporum f.sp. melonis race 1 determined by inoculation tests and molecular markers

Screening of genotypes of melon ( Cucumis melo ) for resistance to wilt caused by Fusarium oxysporum f.sp. melonis is often characterized by wide variability in their responses to inoculation, even under carefully controlled conditions. The variability at the seedling stage of 17 genotypes susceptible to race 1 was examined in growth-chamber experiments. Disease incidence varied from 0 to 100% in a genotype-dependent manner. Using four combinations of light (60 and 90  µ E m⁻² s⁻¹) and temperatures of (27 and 31°C), only light intensity showed a statistically significant effect. Marker-assisted selection for fusarium resistance breeding using cleaved amplified polymorphic sequence (CAPS) and sequence-characterized amplified region (SCAR) markers were compared using a single set of genotypes that included 24 melon accessions and breeding lines whose genotype regarding the Fom-2 gene was well characterized. The practical value of the markers for discriminating a range of genotypes and clarifying the scoring of phenotypes was also tested using a segregating breeding population which showed codominant SCAR markers to be useful in marker-assisted selection.     

Variations in vernalization requirements among ecotypes of Festuca hallii

Plains rough fescue (Festuca hallii [Vasey] Piper) is an important forage grass species in the Northern Great Plains of Canada. Its seed is in demand for forage production and habitat restoration, but erratic seed production limits supply. A comprehensive understanding of factors influencing flowering and seed production in this species is needed. This study evaluated the morphological and phenological variation among six ecotypes of F. hallii from Saskatchewan and Manitoba. Seeds were germinated, and seedlings were grown in the field and then transplanted to a greenhouse in November. Plants not flowering in the greenhouse were vernalized under 5°C and 8-h light for 11 weeks. In a separate experiment, plants were subjected to temperature regimes of 15/5°C, 10/0°C and 5/−5°C with day-length treatments of 12 h, 8 h and a gradually changing daylength from 12 to 8 h respectively. This study demonstrated the existence of considerable variation in morphological and phenological characteristics, and in growth and vernalization requirements among ecotypes of F. hallii. Vernalization requirements were not met for the ecotype from the Moist Mixed Grassland Ecoregion when it was grown under common conditions, whereas ecotypes from other ecoregions were vernalized in at least one of the 2 years in the field experiment. Northern ecotypes tended to flower earlier after artificial vernalization treatments. Overall, 15/5 to 5/−5°C d per night temperature regimes with photoperiods between 12 and 8 h were effective in inducing flowering. The seed source of F. hallii should be regarded as an important consideration affecting its use, both for habitat restoration and for forage production.     

Interaction of Crop Load and Irrigation on Yield,Fruit Size,Color and Stem-end Splitting Ratio of Apple c.v. ‘Gala,Galaxy’

In this study, the interaction between crop load and irrigation level on yield, fruit size, skin color and stem-end splitting fruit ratio in the apple cultivar ‘Gala, Galaxy’ grafted on rootstock M9 were investigated. Six irrigation programs were applied during the whole growth season: deficit irrigation (rates of 0.25 kc, 0.50 kc, 0.75 kc), full irrigation (rate of 1.00 kc), excess irrigation (rate of 1.25 kc) and non-irrigation (rates of 0.00 kc of “Class A” pan evaporation coefficient). Four crop loads in each irrigation application were performed by hand thinning after the June drop as a- a low crop load (3 fruits cm^?2 TCA), b- a medium crop load (5 fruits cm^?2 TCA), c- a heavy crop load (7 fruits cm^?2 TCA), and d- an un-thinned crop load (>?7 fruits cm^?2 TCA). The total tree yield increased with crop load and irrigation levels. Fruit size was significantly increased by the low crop load. Irrigation increased the fruit size compared to non-irrigation treatment. Further 0.75 kc, 1.00 kc and 1.25 kc irrigation treatments significantly increased the fruit length. Irrigation reduced the fruit flesh firmness. While the low crop load increased the skin red color, it decreased the fruit skin brightness. The yellowness of skin decreased with increasing in the irrigation amount. Irrigation reduced the skin brightness and yellowness, but it increased red color. Crop load and irrigation significantly affected the stem-end splitting fruit ratio. While the splitting fruit ratio increased with a decrease in the crop load, it decreased with an increase in irrigation amount, relatively. Consequently, the low and medium crop load treatments would be beneficial to increase the ratio of marketable fruits without any significant losses in yield for ‘Gala’ apple, especially under 0.75 kc deficit irrigation treatment.     

Immunogene expression in head kidney and spleen of common carp (<Emphasis Type="Italic">Cyprinus carpio</Emphasis> L.) following thermal stress and challenge with Gram-negative bacterium, <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

To evaluate the effect of thermal and microbial stress on the immune response of common carp (Cyprinus carpio L.), relative mRNA expression level of pro-inflammatory cytokines [tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1β] and other genes related to immune or stress response [inducible nitric oxide synthase (iNOS), heat shock protein 70 (Hsp70), superoxide dismutase one (SOD1), and glucocorticoid receptor (GR)] was measured by quantitative PCR (qPCR). In addition, total protein and total immunoglobulin level in blood plasma of experimental common carp was also assayed. All the above parameters were estimated 24 h post-challenge with Gram-negative bacterium, Aeromonas hydrophila. Common carp (54.89?±?6.90 g) were initially exposed to 20 °C (control group) and 30 °C (thermal stress group) water temperature for 30 days, followed by experimental challenge with 2.29?×?10⁸ colony forming unit/mL (CFU/mL; LD₅₀ dose) of A. hydrophila. Exposure of fish to thermal stress and subsequently challenge with A. hydrophila significantly (P?<?0.05) increases the IL-1β mRNA expression in head kidney and spleen of common carp by ~?39.94 and ~?4.11-fold, respectively. However, TNF-α mRNA expression in spleen decreased ~?5.63-fold in control fish challenged with A. hydrophila. Thermal stress and challenge with bacterium suppresses the iNOS and GR mRNA expression in spleen of common carp. Moreover, significant (P?<?0.05) increase in total protein content of blood plasma (~?43 mg/g) was evident in fish exposed to thermal stress and challenged with A. hydrophila. In conclusion, our study highlights the importance of elevated temperature stress and microbial infection in differential regulation of expression of several immunogenes in common carp.     

Genotypic Variation for Tolerance to Transient Drought During the Reproductive Phase of Brassica rapa

Brassica rapa L. is a genetically diverse parent species of the allotetraploid species, oilseed rape (B. napus) and a potential source of drought tolerance for B. napus. We examined the effect of a 13‐day drought stress period during the early reproductive phase, relative to a well‐watered (WW) control, on subsequent growth and development in nine accessions of B. rapa and one accession of Brassica juncea selected for their wide morphological and genetic diversity. We measured leaf water potential, stomatal conductance, water use, and leaf and bud temperatures during the stress period and aboveground dry weight of total biomass at maturity. Dry weight of seeds and reproductive tissue were not useful measures of drought tolerance due to self‐incompatibility in B. rapa. The relative total biomass (used as the measure of drought tolerance in this study) of the 10 accessions exposed to drought stress ranged from 47 % to 117 % of the WW treatment and was negatively correlated with leaf‐to‐air and bud‐to‐air temperature difference when averaged across the 13‐day stress period. Two wild‐type (B. rapa ssp. sylvestris) accessions had higher relative total and non‐reproductive biomass at maturity and cooler leaves and buds than other types. We conclude that considerable genotypic variation for drought tolerance exists in B. rapa and cooler leaves and buds during a transient drought stress in the early reproductive phase may be a useful screening tool for drought tolerance.     

β‐hydroxy‐β‐methyl butyrate promotes leucine metabolism and improves muscle fibre composition in growing pigs

The aim of this study was to investigate the effects of excess leucine (Leu) vs. its metabolites α‐ketoisocaproate (KIC) and β‐hydroxy‐β‐methyl butyrate (HMB) on Leu metabolism, muscle fibre composition and muscle growth in growing pigs. Thirty‐two pigs with a similar initial weight (9.55 ± 0.19 kg) were fed 1 of 4 diets for 45 days: basal diet, basal diet + 1.25% L‐Leu, basal diet + 1.25% KIC‐Ca, basal diet + 0.62% HMB‐Ca. Results indicated that relative to the basal diet and HMB groups, Leu and KIC groups exhibited increased Leu concentrations and decreased concentrations of isoleucine, valine and EAAs in selected muscle (p < 0.05) and had lower mRNA levels of MyHC I and higher expression of MyHC IIx/IIb (p < 0.05), and there was no significant difference between the basal and HMB‐supplemented groups. Moreover, the mRNA expression levels of AMPKα and UCP3 were higher but the myostatin mRNA levels were lower in the soleus muscle of the HMB group than those from other groups (p < 0.05). These findings demonstrated that doubling dietary Leu content exerted growth‐depressing effects in growing pigs; dietary KIC supplementation induced muscular branched‐chain amino acid imbalance and promoted muscle toward a more glycolytic phenotype; while dietary HMB supplementation promoted the generation of more oxidative muscle types and increased muscle growth specially in oxidative skeletal muscle, and these effects of HMB might be associated with the AMPKα‐Sirt1‐PGC‐1α axis and mitochondrial biogenesis.     

Immunological characterization of the major outer membrane protein of Haemophilus somnus

Antigens and molecular mass diversity of the Haemophilus somnus major outer membrane protein (MOMP) were investigated. The molecular mass of the MOMP of 53 strains of H. somnus varied from 43 to 33 kDa and four MOMP MAb reactivity patterns were detected in immunoblot analysis and immunodot assay. The molecular mass and MAb reactivity data were used for preliminary grouping of H. somnus strains. Disease strains fell into groups 1 and 3, including two of three Group 3 subgroups, whereas strains from asymptomatic carriers were found in all the four groups and three subgroups. Immunoblot analysis with convalescent phase serum showed strain specific reactivity with MOMPs from three isolates used to reproduce disease in cattle. The reaction with the MOMP was only detectable at dilutions of 1:100 or less, whereas the same convalescent sera showed strong reactivity at dilutions of 1:1000 (or more) with other H. somnus antigens. The data suggest that the bovine immune response to the MOMP during infection is weak and is directed to antigenically variable determinants in a strain-specific manner. This may be important in evaluating the role of the antibody response to MOMPs in protective immunity.