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Filling and emptying of the alimentary tract of meal-fed broiler breeder hens.

For evaluation of the filling and emptying of the alimentary tract, broiler breeder hens were cooped and processed over a 2-d period. Hens were fed at 0600 h on d 1 and after access to feed for 0, 2, 4, and 6 h were placed into coops. Half of the hens from each pen were either immediately processed or were held in coops over-night and processed the following morning, d 2. The alimentary tract was excised from the carcass and then separated and weighed in three segments: the crop, proventriculus and gizzard, and intestines. Hens processed on d 1, after access to feed for only 2 h, had attained maximum intestine weight (176 g), but not until after access to feed for 6 h were peak crop weight (95 g) and peak weight for the proventriculus and gizzard (78 g) attained. Hens processed on d 2 did not differ in crop (12 to 14 g) or intestine (140 to 162 g) weight, but proventriculus and gizzard weights were significantly lower for hens not fed on d 1 prior to cooping (54 g) compared with hens fed on d 1 and cooped after 2, 4, or 6 h (62 to 63 g). However, hens processed on d 2 had proventriculus and gizzard weights that were the same as for those hens processed on d 1 and cooped at 0 h (63 g). Clearance of ingesta from the crop, proventriculus and gizzard, and intestines readily occurred while hens were held overnight without access to water.

Animal Feed↗

Comparison of zinc acetate and propionate addition on gastrointestinal tract fermentation and susceptibility of laying hens to Salmonella enteritidis during forced molt.

Feed deprivation is the most common method used to induce molting and stimulate multiple egg-laying cycles in laying hens for commercial egg production. Unfortunately, an increased risk of Salmonella enteritidis (SE) colonization may result from the use of this method. Methods to stimulate multiple egg-laying cycles without increasing the risk of SE are needed. In each of 3 experiments, hens over 50 wk of age were divided into groups of 12 and placed in individual laying cages. One week before dietary changes, hens were put on an 8L:16D photoperiod that continued for the 9-d experimental period. Hens in all treatments were challenged orally with 10(4) cfu of SE on the fourth day. Treatments were full fed hens (nonmolted, NM), nonfed hens (molted, M), a zinc acetate diet (ZAC), and a zinc propionate diet (ZPR). The zinc diets contained 10,000 mg of zinc per kilogram of diet. Body weight losses were significantly higher in the M, ZPR, and ZAC treatments than in the NM treatment. Crop lactic acid decreased more in M, ZPR, and ZAC treatments than in NM hens in trial 2. Crop pH was significantly (P < 0.05) lower in NM hens than in M, ZAC, and ZPR hens in trial 2. Although cecal individual or total volatile fatty acids (VFA), and lactic acid were not significantly (P > 0.05) different between NM hens and M, ZAC and ZPR hens in trial 1, lactic acid was significantly (P < 0.05) higher in NM hens than in M, ZAC and ZPR hens (trial 2), and cecal total VFA were lower in M hens than in NM, ZAC and ZPR hens (trial 3). Colonization of SE in the crop and ceca was higher in the M and ZPR hens (trials 1 and 2). Liver, spleen, or ovary invasion by SE was higher in the M and ZPR hens (trials 1 and 2) than in NM hens. At the zinc concentration used in these studies, the zinc dietary regimens may be effective for reducing the risk of SE during induced molt.

Animals↗

RAPD and other PCR-based analyses of plant genomes using DNA extracted from small leaf disks.

A nondestructive, early DNA diagnostic system to implement marker-assisted selection in plant breeding programs has been developed. The main components of the system are a rapid and simple DNA microextraction method and fast DNA polymorphism analyses based on site-specific or arbitrary DNA amplification. A small disk (5 mm diameter) is collected from one cotyledon or the first leaf of a young seedling using a common paper punch. Disruption of plant tissues is done by enzymatic digestion of cell walls. This ensures protection from sample-to-sample contamination and uniform DNA yield. DNA isolated from the resulting protoplasts is sufficient to perform a minimum of five and a maximum of 20 PCR reactions/sample. Total DNA, nuclear DNA, and RNA can be analyzed selectively. The system has been tested successfully with eight major crops. Amplification products generated with DNA prepared with this quick procedure are equivalent to those obtained from CsCl-purified DNA. Up to 120 plants can be treated in 2 days and the procedure lends itself to automation. Potential applications in plant breeding will be discussed.

Base Sequence↗

The nicotinic acetylcholine receptor gene family of the honey bee, Apis mellifera.

Nicotinic acetylcholine receptors (nAChRs) mediate fast cholinergic synaptic transmission and play roles in many cognitive processes. They are under intense research as potential targets of drugs used to treat neurodegenerative diseases and neurological disorders such as Alzheimer's disease and schizophrenia. Invertebrate nAChRs are targets of anthelmintics as well as a major group of insecticides, the neonicotinoids. The honey bee, Apis mellifera, is one of the most beneficial insects worldwide, playing an important role in crop pollination, and is also a valuable model system for studies on social interaction, sensory processing, learning, and memory. We have used the A. mellifera genome information to characterize the complete honey bee nAChR gene family. Comparison with the fruit fly Drosophila melanogaster and the malaria mosquito Anopheles gambiae shows that the honey bee possesses the largest family of insect nAChR subunits to date (11 members). As with Drosophila and Anopheles, alternative splicing of conserved exons increases receptor diversity. Also, we show that in one honey bee nAChR subunit, six adenosine residues are targeted for RNA A-to-I editing, two of which are evolutionarily conserved in Drosophila melanogaster and Heliothis virescens orthologs, and that the extent of editing increases as the honey bee lifecycle progresses, serving to maximize receptor diversity at the adult stage. These findings on Apis mellifera enhance our understanding of nAChR functional genomics and provide a useful basis for the development of improved insecticides that spare a major beneficial insect species.

Alternative Splicing↗

Conservation of Arabidopsis flowering genes in model legumes.

The model plants Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa) have provided a wealth of information about genes and genetic pathways controlling the flowering process, but little is known about the corresponding pathways in legumes. The garden pea (Pisum sativum) has been used for several decades as a model system for physiological genetics of flowering, but the lack of molecular information about pea flowering genes has prevented direct comparison with other systems. To address this problem, we have searched expressed sequence tag and genome sequence databases to identify flowering-gene-related sequences from Medicago truncatula, soybean (Glycine max), and Lotus japonicus, and isolated corresponding sequences from pea by degenerate-primer polymerase chain reaction and library screening. We found that the majority of Arabidopsis flowering genes are represented in pea and in legume sequence databases, although several gene families, including the MADS-box, CONSTANS, and FLOWERING LOCUS T/TERMINAL FLOWER1 families, appear to have undergone differential expansion, and several important Arabidopsis genes, including FRIGIDA and members of the FLOWERING LOCUS C clade, are conspicuously absent. In several cases, pea and Medicago orthologs are shown to map to conserved map positions, emphasizing the closely syntenic relationship between these two species. These results demonstrate the potential benefit of parallel model systems for an understanding of flowering phenology in crop and model legume species.

Arabidopsis↗

Processing and secretion of a virally encoded antifungal toxin in transgenic tobacco plants: evidence for a Kex2p pathway in plants.

Ustilago maydis is a fungal pathogen of maize. Some strains of U. maydis encode secreted polypeptide toxins capable of killing other susceptible strains of U. maydis. We show here that one of these toxins, the KP6 killer toxin, is synthesized by transgenic tobacco plants containing the viral toxin cDNA under the control of a cauliflower mosaic virus promoter. The two components of the KP6 toxin, designated alpha and beta, with activity and specificity identical to those found in toxin secreted by U. maydis cells, were isolated from the intercellular fluid of the transgenic tobacco plants. The beta polypeptide from tobacco was identical in size and N-terminal sequence to the U. maydis KP6 beta polypeptide. Processing of the KP6 preprotoxin in U. maydis requires a subtilisin-like processing protease, Kex2p, which is present in both animal and fungal cells and is required for processing of (among other things) small secreted polypeptide hormones and secreted toxins. Our findings present evidence for Kex2p-like processing activity in plants. The systemic production of this viral killer toxin in crop plants may provide a new method of engineering biological control of fungal pathogens in crop plants.

Cloning, Molecular↗

Efficient insertional mutagenesis in rice using the maize En/Spm elements.

We have developed a novel system for insertional mutagenesis in rice (Oryza sativa) based on the maize (Zea mays) enhancer/suppressor mutator (En/Spm) element. In this system, a single T-DNA construct with Spm-transposase and the non-autonomous defective suppressor mutator (dSpm) element is used in conjunction with green fluorescent protein (GFP) and Discosoma sp. Red Fluorescence Protein (DsRed) fluorescent markers to select unlinked stable transpositions of dSpm. Using this system, we could demonstrate high frequencies of unlinked germinal transposition of dSpm in rice. Analysis of dSpm flanking sequences from 353 stable insertion lines revealed that the dSpm insertions appear to be widely distributed on rice chromosomes with a preference for genic regions (70%). The dSpm insertions appear to differ from Activator-Dissociation (Ac-Ds) elements in genomic distribution and exhibit a greater fraction of unlinked transpositions when compared with Ds elements. The results obtained in this study demonstrate that the maize En/Spm element can be used as an effective tool for functional genomics in rice and can complement efforts using other insertional mutagens. Further, the efficacy of the non-invasive fluorescence-based selection system is promising for its application to other crops.

Chromosomes, Plant↗

Enhanced levels of methionine and cysteine in transgenic alfalfa (Medicago sativa L.) plants over-expressing the Arabidopsis cystathionine gamma-synthase gene.

With the aim of increasing the methionine level in alfalfa (Medicago sativa L.) and thus improving its nutritional quality, we produced transgenic alfalfa plants that expressed the Arabidopsis cystathionine gamma-synthase (AtCGS), the enzyme that controls the synthesis of the first intermediate metabolite in the methionine pathway. The AtCGS cDNA was driven by the Arabidopsis rubisco small subunit promoter to obtain expression in leaves. Thirty transgenic plants were examined for the transgene protein expression, and four lines with a high expression level were selected for further work. In these lines, the contents of methionine, S-methylmethionine (SMM), and methionine incorporated into the water-soluble protein fraction increased up to 32-fold, 19-fold, and 2.2-fold, respectively, compared with that in wild-type plants. Notably, in these four transgenic lines, the levels of free cysteine (the sulphur donor for methionine synthesis), glutathione (the cysteine storage and transport form), and protein-bound cysteine increased up to 2.6-fold, 5.5-fold, and 2.3-fold, respectively, relative to that in wild-type plants. As the transgenic alfalfa plants over-expressing AtCGS had significantly higher levels of both soluble and protein-bound methionine and cysteine, they may represent a model and target system for improving the nutritional quality of forage crops.

Journal Article↗

Characterization of the bacterial flora associated with root systems of Pinus contorta var. latifolia.

Root systems of young and mature lodgepole pine (Pinus contorta Dougl. var. latifolia Englem.) were removed from forest stands and the associated aerobic bacterial flora isolated. Characterization of rhizoplane and control soil isolates from these tree root systems demonstrated differences from that reported for agricultural crops. Ammonifying, proteolytic, and amylolytic organisms were proportionately reduced within the rhizoplane. The rhizoplane organisms grew more slowly than the control soil isolates, although they responded in greater numbers to the addition of an amino acid supplement to the growth media. The rhizoplane organisms also showed an increased ability to solubilize phosphate. The chitinolytic organisms were suppressed within the rhizoplane of the mature tree but were stimulated by the young trees. With this exception, the rhizoplane microflora of older and younger trees were similar.

Ammonia↗

Stimulatory effect of pigeon milk growth factor on protein and nucleic acid synthesis in neonate mice.

Effect of subcutaneously administered pigeon milk-derived growth factor (PMGF) on protein and nucleic acid content of various body organs was studied in neonate mice. Although the body and organ weights of PMGF-treated mice did not differ from those of controls, the weight of testes was significantly higher in the former than the latter. The treated animals had significantly greater content of protein (in liver and skin), DNA (in liver, lung, stomach, duodenum, ileum and rectum) and RNA (in liver, stomach, ileum and skin). The protein/DNA ratio was lower in many organs of PMGF-treated mice. These results indicate that the biological properties of PMGF bear semblance to those of epidermal growth factor isolated from mouse salivary gland and human milk.

Animals↗

Optimized dairy grazing systems in the northeast United States and New Zealand. II. System analysis.

Factors that optimize milk production from Northeast United States and New Zealand grazing systems are compared using a linear programming model. The objective function maximized gross margin per hectare of land farmed. The experimental design compared the optimum characteristics of each system over a range of milk prices. The Northeast has a shorter grazing season and lower cropping costs than New Zealand. The optimum pasture area was 49% of the farm for Northeast systems. Gross margins declined rapidly above 55% or below 36% pasture area. The optimum stocking rate was 1.13 cows/ha, or 2.3 cows/ha of pasture. Optimum per cow production was higher for Northeast [7105 kg of fat-corrected milk (FCM)] than New Zealand (5710 kg of FCM) systems. This was related to lower grain relative to milk prices in the Northeast. New Zealand, all-pasture systems gave the lowest cost per unit of milk but also gave the lowest gross margin across all milk price scenarios. The best use of purchased feed in New Zealand systems was to support increased stocking rate rather than per cow production. Optimum grazing management practices were similar for supplemented New Zealand and Northeast systems. All-pasture New Zealand systems are characterized by short lactations and long autumn rotations to transfer pasture in situ for winter feeding. Higher costs per unit of milk produced will be an inevitable consequence of maximizing gross margin at high milk prices in New Zealand systems.

Agriculture↗

Seasonal phenology of Bactrocera minax (Diptera: Tephritidae) in western Bhutan.

The Chinese citrus fruit fly, Bactrocera (Tetradacus) minax (Enderlein), is one of the major citrus pests in Bhutan and can cause >50% mandarin (Citrus reticulata Blanco) fruit drop. As part of the development of a management strategy for the fly in mandarin orchards, population monitoring and experimental manipulations were carried out to determine: (i) adult emergence period; (ii) adult phenology patterns; (iii) period of crop susceptibility; and (iv) period from fruit drop to pupation. In western Bhutan, adult flies emerge from the overwintering pupal stage in late April/early May. Most flies are mature by the end of May and it is inferred that mating occurs at this time: from the beginning of June males rapidly disappear from the population and by mid- to late June are rare or absent from traps. Mature females are present in the mandarin crop at the beginning of June, but very little oviposition occurs until mid-June, while most damage has occurred by mid-July. Initiation of oviposition into mandarins is almost certainly linked to crop phenology. Adult flies disappear from the orchard system during August. After fruit drop, larvae were recorded leaving the fruit to pupate within 13 days. The use of early to mid-season protein bait sprays and/or targeted use of systemic insecticides during the one month oviposition period, plus the removal of fallen fruit once every 10 days, are recommended as control strategies.

Animals↗

[Electromyographic correlations of ingesta movements in the proximal digestive tract of the hen].

Digesta transit along the oesophagus, crop and stomach has been determined by the migration of spike bursts correlated with cineradiographic observations in hens fitted with chronically implanted electrodes. The passage of digesta along the oesophagus corresponded to the propagation of isolated spike bursts. During crop filling, the activity was weak, its emptying being mediated by migrating repetitive spike bursts. A cyclically recurring electrical activity was noted for the glandular and muscular stomachs and the duodenum; their motility is described in relation to digesta movements.

Animals↗

Plant glutathione S-transferases and herbicide detoxification.

Glutathione S-transferases (GSTs) are a family of multifunctional enzymes involved in the metabolism of xenobiotics and reactive endogenous compounds. The interest in plant GSTs may be attributed to their agronomic value, since it has been demonstrated that glutathione conjugation for a variety of herbicides is the major resistance and selectivity factor in plants. The structure of the Arabidopsis thaliana isoenzyme, the first plant GST whose structure has been solved, may serve as a model system for the understanding of herbicide selectivity in crops.

Amino Acid Sequence↗

Status of public health--Democratic People's Republic of Korea, April 1997.

During 1995 and 1996, severe flooding in the Democratic People's Republic of Korea (i.e., North Korea (DPRK) (1990 population: 22 million)) (Figure 1) caused 186 deaths, dislocated approximately 550,000 persons from their homes, and caused damage to an estimated 1.2 million metric tons of crops (12% of total production). In combination with systemic economic problems in DPRK, these natural disasters have been associated with reports of a severe, ongoing food shortage and increased risks to public health. To assist in targeting humanitarian aid, in April 1997, the U.S. Agency for International Development's Office of U.S. Foreign Disaster Assistance requested CDC to conduct an onsite assessment of the public health status and needs of the DPRK. This report summarizes findings of the assessment, which indicate a recent substantial decline in the health and nutritional status in DPRK.

Disasters↗

The GP problem: quantifying gene-to-phenotype relationships.

In this paper we refer to the gene-to-phenotype modeling challenge as the GP problem. Integrating information across levels of organization within a genotype-environment system is a major challenge in computational biology. However, resolving the GP problem is a fundamental requirement if we are to understand and predict phenotypes given knowledge of the genome and model dynamic properties of biological systems. Organisms are consequences of this integration, and it is a major property of biological systems that underlies the responses we observe. We discuss the E(NK) model as a framework for investigation of the GP problem and the prediction of system properties at different levels of organization. We apply this quantitative framework to an investigation of the processes involved in genetic improvement of plants for agriculture. In our analysis, N genes determine the genetic variation for a set of traits that are responsible for plant adaptation to E environment-types within a target population of environments. The N genes can interact in epistatic NK gene-networks through the way that they influence plant growth and development processes within a dynamic crop growth model. We use a sorghum crop growth model, available within the APSIM agricultural production systems simulation model, to integrate the gene-environment interactions that occur during growth and development and to predict genotype-to-phenotype relationships for a given E(NK) model. Directional selection is then applied to the population of genotypes, based on their predicted phenotypes, to simulate the dynamic aspects of genetic improvement by a plant-breeding program. The outcomes of the simulated breeding are evaluated across cycles of selection in terms of the changes in allele frequencies for the N genes and the genotypic and phenotypic values of the populations of genotypes.

Computer Simulation↗

Reproductive performance of crossbred dairy cows in different production systems in the Central Highlands of Ethiopia.

The study was conducted to examine the fertility status of crossbred dairy cows in mixed crop-livestock production (MCLP), market-oriented specialized dairy production (MSDP) and urban dairy production (UDP) systems, including the Holetta Agricultural Research Center (HARC). Data on general farm management variables and reproductive histories were collected from study farms by questionnaire and from individual cow records. Age at first service and age at first calving were 29.58 months (n = 424) and 40.6 months (n = 348), respectively. Cows managed under UDP were younger at first service and at first calving (p<0.05). The mean intervals from calving to first service and to conception were 141.98 days (n = 284) and 185.02 days (n = 219), respectively. The mean calving interval for cows was 551.82 days (n = 258). Cows in MCLP had longer intervals from calving to first service and to conception and longer calving intervals than those managed under MSDP, UDP and HARC. First service conception rate (43.42%), number of services per conception (1.75) and pregnancy rate (79.29%) did not differ significantly between production systems. Reproductive performance was best in UDP followed by HARC and MSDP. The difference between MCLP and the rest points to particular difficulties in that system. To improve reproductive performance and economic benefit, there should be conservative stocking rate, sensible year-round feeding, a herd health plan, and sustainable extension service.

Animals↗