Egg production, faecal pH and microbial population, small intestine morphology, and plasma and yolk cholesterol in laying hens given liquid metabolites produced by Lactobacillus plantarum strains

1. Various dosages of metabolite combinations of the Lactobacillus plantarum RI11, RG14 and RG11 strains (COM456) were used to study the egg production, faecal microflora population, faecal pH, small intestine morphology, and plasma and egg yolk cholesterol in laying hens. 2. A total of 500 Lohmann Brown hens were raised from 19 weeks to 31 weeks of age. The birds were randomly divided into 5 groups and fed on various treatment diets: (i) basal diet without supplementation of metabolites (control); (ii) basal diet supplemented with 0·3% COM456 metabolites; (iii) basal diet supplemented with 0·6% COM456 metabolites; (iv) basal diet supplemented with 0·9% COM456 metabolites; and (v) basal diet supplemented with 1·2% COM456 metabolites. 3. The inclusion of 0·6% liquid metabolite combinations, produced from three L. plantarum strains, demonstrated the best effect in improving the hens' egg production, faecal lactic acid bacteria population, and small intestine villus height, and reducing faecal pH and Enterobacteriaceae population, and plasma and yolk cholesterol concentrations. 4. The metabolites from locally isolated L. plantarum are a possible alternative feed additive in poultry production.     

Development of an ELISA using a recombinant 41 kDa partial protein (P45N') for the detection of Riemerella anatipestifer infections in ducks

Riemerella anatipestifer, a gram-negative bacillus, is the causative agent of duck septicemia, a disease which could incur much economic loss in the duck industry. An indirect enzyme-linked immunosorbent assay (ELISA) has been developed to facilitate early detection of R. anatipestifer infection in ducks. The antigen used was a recombinant 41 kDa N-terminal fragment (rP45N') of a newly characterized R. anatipestifer potential surface protein, P45, which was expressed in Escherichia coli as an N-terminal GST fusion protein. The rP45N'-based ELISA successfully detected P45 antibodies in the sera of 20 ducks immunized with bacterin preparations of R. anatipestifer serotypes 1, 10 15, 19 and the ATCC11845 strain. Antibodies to P45 were also detected in the sera of 25% (75/296) of White Pekin ducks which were imported into Singapore from three different farms. Successful discrimination was obtained between sera from infected ducks and that of specific-pathogen free ducks (p<0.01). The rP45N'-GST antigen did not cross-react with antibodies in sera from guinea pigs which were infected with other gram-negative and gram-positive bacterial pathogens, including Aeromonas hydrophila, Citrobacter freundii, E. coli, Klebsiella pneumoniae, Pastuerella multocida, Proteus mirabilis, Salmonella spp., Serratia maccescens, Shigella sonnei and Yersinia enterocolitica. In addition, the DNA sequence encoding P45 was detected in R. anatipestifer serotypes 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 14, 15, 16, 17, 18, 19 and the ATCC11845 strain, suggesting that P45 is probably also universally expressed in these R. anatipestifer serotypes. Thus, the ELISA described is applicable to the detection of R. anatipestifer infection in ducks.     

Prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in goats slaughtered in the public slaughterhouse of Patos city, Paraíba State, Northeast region of Brazil

The prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies was investigated in goats slaughtered in the public slaughterhouse of Patos, State of Paraíba, Northeast region of Brazil, and possible associations between sex of the animals and antibody prevalence were verified. Three-hundred six blood samples from goats collected before slaughter by jugular venopuncture were used. For the serologic diagnosis of T. gondii and N. caninum, the indirect fluorescent-antibody test (IFAT) with cut-off values 64 and 50, respectively, was carried out. The prevalence of anti-T. gondii antibodies was 24.5% [95% CI=19.8-29.7%] with titers ranging from 64 to 4096, and anti-N. caninum antibodies was 3.3% (95% CI=1.6-5.9%) with titers ranging from 50 to 400. There were no associations between sex of animals and prevalence of anti-T. gondii and anti-N. caninum antibodies.     

A simple,rapid method to isolate salt glands for three-dimensional visualization,fluorescence imaging and cytological studies

Background  

Some plants inhabiting saline environment remove salts via the salt glands embedded in the epidermal tissues. Cytological studies of salt glands will provide valuable information to our understanding of the secretory process. Previous studies on salt gland histology relied mainly on two-dimensional microscopic observations of microtome sections. Optical sectioning properties of confocal laser scanning microscope offer alternative approach for obtaining three-dimensional structural information of salt glands. Difficulty in light penetration through intact leaves and interference from neighbouring leaf cells, however, impede the acquiring of good optical salt gland sections and limit its applications in salt gland imaging. Freeing the glands from adjacent leaf tissues will allow better manipulations for three-dimensional imaging through confocal laser scanning microscopy.     

Immunoassay for wheat processing quality: utilization of a sandwich assay incorporating an immobilized single-chain fragment.

A single-chain fragment (scFv) was engineered from a monoclonal antibody to high molecular weight glutenin subunits (HMW-GS), wheat flour polypeptides that play a major role in determining the mixing- and extension strength-related properties of dough and its subsequent baking performance. The scFv was expressed in a thioredoxin mutant Escherichia coli strain that allows disulfide bond formation in the cytoplasm and incorporated into a diagnostic test for wheat quality. Although the scFv lacks the more highly conserved antibody constant regions usually involved with immobilization, it was able to be directly immobilized to a polystyrene microwell solid phase without chemical or covalent modification of the protein or solid phase and utilized as a capture antibody in a double-antibody (two-site) immunoassay. In the sandwich assay, increasing HMW-GS concentrations produced increasing assay color, and highly significant correlations were obtained between optical densities obtained in the ELISA using the scFv and the content of large glutenin polymers in flours as well as measures of dough strength as measured by resistance to dough extension in rheological testing. The assay using the scFv was able to be carried out at lower flour sample extract dilutions than that required for a similar assay utilizing a monoclonal capture antibody. This research shows that engineered antibody fragments can be utilized to provide superior assay performance in two-site ELISAs over monoclonal antibodies and is the first application of an engineered antibody to the analysis of food processing quality.     

The effects of pharmacological levels of zinc,diet acidification,and dietary crude protein on growth performance in nursery pigs

This experiment was conducted to evaluate potential replacements for pharmacological levels of Zn (provided by Zn oxide), such as diet acidification (sodium diformate) and low dietary crude protein (CP: 21 vs 18%) on nursery pig performance and fecal dry matter (DM). A total of 360 weaned pigs (Line 200 × 400, DNA, Columbus, NE; initially 5.90 ± 0.014 kg) were used in a 42-d growth study. Pigs were weaned at approximately 21 d of age and randomly assigned to pens (five pigs per pen). Pens were then allotted to one of eight dietary treatments with nine pens per treatment. Experimental diets were fed in two phases: phase 1 from weaning to day 7 and phase 2 from days 7 to 21, with all pigs fed the same common diet from days 21 to 42. The eight treatment diets were arranged as a 2 × 2 × 2 factorial with main effects of Zn (110 mg/kg from days 0 to 21 or 3,000 mg/kg from days 0 to 7, and 2,000 mg/kg from days 7 to 21), diet acidification, (without or with 1.2% sodium diformate), and dietary CP (21% or 18%, 1.40% and 1.35% in phases 1 and 2 vs. 1.20% standardized ileal digestible Lys, respectively). Fecal samples were collected weekly from the same three pigs per pen to determine DM content. No 2- or 3-way interactions (P > 0.05) were observed throughout the 42-d study for growth performance; however, there was a Zn × acidifier × CP interaction (P < 0.05) for fecal DM on day 7 and for the overall average of the six collection periods. Reducing CP without acidification or pharmacological levels of Zn increased fecal DM, but CP had little effect when ZnO was present in the diet. From days 0 to 21, significant (P < 0.05) main effects were observed where average daily gain (ADG) and gain:feed (G:F) increased for pigs fed pharmacological levels of Zn, sodium diformate, or 21% CP (P < 0.065). In the subsequent period (days 21 to 42) after the experimental diets were fed, there was no evidence of difference in growth performance among treatments. Overall (days 0 to 42), main effect tendencies were observed (P < 0.066) for pigs fed added Zn or sodium diformate from days 0 to 21, whereas pigs fed 21% CP had greater G:F than those fed 18% CP. Pig weight on day 42 was increased by adding Zn (P < 0.05) or acidifier (P < 0.06) but not CP. In summary, none of the feed additives had a major influence on fecal DM, but dietary addition of pharmacological levels of Zn or sodium diformate independently improved nursery pig performance.     

Case studies of spinal deformities in ornamental koi,Cyprinus carpio L.

This is a study of vertebral deformities in ornamental koi based on computed radiography and skeletons cleaned by dermestid beetles (Dermestes maculatus). All koi developed gradual onset of swimming abnormalities as adults. Extensive intervertebral osteophyte formation correlated with age of fish and was associated with hindquarter paresis in one koi. Vertebral compression and fusion were the most common spinal deformities occurring at multiple sites, similar to findings in other farmed fish. Site‐specific spinal deformities were thought to develop due to differences in swimming behaviour and rates of vertebral growth. One koi had offspring with spinal deformities. Spinal deformities are significant problems in both European and Australian food fish hatcheries. The heritability of vertebral deformities in farmed fish is reportedly low unless there is concurrent poor husbandry or nutritional deficiencies. The specific aetiologies for vertebral deformities in koi in this study could not be ascertained. Current knowledge on spinal deformities in the better studied European food fish species suggests multifactorial aetiologies. Future research should include prospective longitudinal studies of larger numbers of koi from hatch and consideration of all potential risk factors such as husbandry, nutrition, temperature, photoperiod and genetics.     

Molecular characterization and analysis of somaclonal variation in chrysanthemum cultivars using RAPD markers

Molecular characterization using RAPD analysis was carried out in eight cut flowers and two pot plant cultivars of chrysanthemum. Three of them (‘Refocus’, ‘Red Reagan’, and ‘Sheena Select’) were established in vitro and the occurrence of somaclonal variation was studied using the same molecular technique. Two induction media (MS + 0.1 mg l⁻¹ NAA + 0.1 mg l⁻¹ BA, and MS + 2.0 mg l⁻¹ IAA + 0.5 mg l⁻¹ Kinetin), and two proliferation media (MS + 0.1 mg l⁻¹ NAA + 0.2 mg l⁻¹ BA, and MS + 4.0 mg l⁻¹ IAA + 2.0 mg l⁻¹ Kinetin) were employed in order to evaluate the effect of the medium composition in the shoots’ stability. Likewise, the effect of the culture age was considered in assessing genetic stability. Monthly subcultures were carried out, identifying the origin and history of the shoots, throughout a nine-month proliferation period followed by acclimatization. Molecular markers were obtained in every subculture cycle and from the acclimatized plants. Only one shoot from the 7th subculture of the cultivar ‘Refocus’ showed a different band pattern. The use of RAPD for chrysanthemum cultivar characterization and somaclonal variation detection is discussed.