Can Bacteriotherapy Using Commercially Available Probiotics, Prebiotics, and Organic Acids Ameliorate the Symptoms Associated With Runting-Stunting Syndrome in Broiler Chickens?

Runting-stunting syndrome (RSS) in poultry has been known for more than 40 years, but the precise etiology remains unknown and a licensed vaccine is consequently not currently available. In order to mitigate the symptoms associated with RSS, a series of experiments was performed to investigate whether a combined bacteriotherapeutic treatment consisting of probiotics, prebiotics, and organic acids could influence the outcome of this disease. Initially two groups of commercial broiler chickens were either left uninoculated or inoculated with filtrate from homogenized intestines of RSS-affected broiler chickens. One group from each of these two challenge groups was treated, with a bacteriotherapeutic regimen. After 12 days chickens were euthanatized, the body weight was measured, and duodenal lesions were enumerated. Five consecutive broiler chicken flocks were then raised either on litter from RSS-affected birds or on fresh wood shavings. Treatment had no beneficial effect on the number and severity of intestinal lesions. There appeared to be a significant build-up of RSS agent(s) in poultry litter, with each consecutive flock placement, independent of bacteriotherapeutic treatment, as more individuals exhibited intestinal lesions on built-up litter in RSS-affected houses (28.9% vs. 44%). While treatment did not appear to consistently reduce intestinal lesions, it did significantly improve the mean body weights (P<0.05) and uniformity of 12-day-old chickens placed on reused litter in houses in which RSS-infected birds were previously raised. A combination of litter management and bacteriotherapy may be needed to ameliorate the adverse effects of RSS on intestinal health and body weight in broiler chickens.

Metastatic melanomas in young broiler chickens (Gallus gallus domesticus).

Four young broiler chickens affected by multiple melanotic tumors are described. Grossly, there were multiple tumors composed of melanocytes within the skin, skeletal muscle, and multiple visceral organs. Tumors ranged from flattened macules to masses that extensively replaced viscera. Microscopically, melanocytes were often well pigmented, and while there was moderate nuclear anisokaryosis, mitotic rates were low. Immunohistochemical staining of some melanomas with antibodies to S100 proteins, Melan-A, vimentin, or neuron-specific enolase after bleaching of tumor cells with potassium permanganate revealed lack of immunostaining of tumor cells with antibodies to S100, strong positive staining of tumor cells for neuron-specific enolase, moderate staining with antibodies to vimentin, and faint staining for Melan-A. Only neuron-specific enolase staining was evident in unbleached tumor cells. Attempts to identify exogenous avian leukosis viruses in these tumors were unsuccessful.

Current status of routine post-mortem CT in Melbourne, Australia.

In May 2005, a multidetector computed tomography (CT) scanner was installed into the mortuary of the Victorian Institute of Forensic Medicine (VIFM). Since that time most biological material admitted to the institute has been scanned. This article provides an overview of the experience gained and difficulties encountered by participants in this project. Discussion is presented on the incorporation of routine CT imaging into autopsy procedures, application of CT in forensic research, and how we believe the use of cross-sectional imaging will evolve in forensic investigation of the deceased.

Floor pen study to evaluate the serological response of broiler breeders after vaccination with ts-11 strain Mycoplasma gallisepticum vaccine.

To determine the Mycoplasma gallisepticum (MG) rapid serum plate agglutination (RSPA) test response of broiler breeders after ts-11 strain vaccination, 55 Cobb pullets derived from a nonvaccinated, MG-negative, commercial, broiler breeder grandparent flock were monitored from 8 to 20 wk of age (over a 12-wk trial period). To evaluate the effect of lateral spread of the ts-11 vaccine strain on RSPA test results from commingled and adjacently penned birds, treatment groups included (A) birds vaccinated with ts-11strain MG at 8 wk of age, (B) commingled nonvaccinates in the same pen as the vaccinated birds, (C) nonvaccinates in a second pen separated from the first pen by a distance of 2 m, and (D) birds vaccinated with ts-11 strain MG at 8 wk of age and kept in a separate room. Rapid serum plate agglutination tests were performed once a week for 6 wk and then every 2 wk for 6 more wk, postvaccination. A polymerase chain reaction (PCR) assay specific fbr ts-11 strain MG was used to confirm vaccination, and a second PCR specific for non-ts-11 strain MG was used to confirm the absence of field infection. Seroconversion was first detected by the RSPA test 2 wk postvaccination and attained maximum positive rates of 58% at 12 wk postvaccination in treatment A and 60% at 8 wk postvaccination in treatment D. Seroconversion rates in nonvaccinated, commingled pullets was 10% at 5 wk and 30% at 12 wk after the vaccination of pen mates. The ts-11-specific PCR detected the vaccine strain in 80%-100% of the vaccinated birds 2 wk after vaccination. One of 15 nonvaccinated birds penned 2 m from vaccinated birds yielded ts-11 by PCR assay 12 wk after vaccination, which indicates that the spread of ts-11 over short distances may be possible in situations in which there is a common caretaker. PCR on tracheal swabs taken 12 wk postvaccination detected ts-l1 in 50% and 60% of the vaccinated birds in treatments A and D, respectively; in 30% of the commingled nonvaccinates; and in 6.6% of the separately penned nonvaccinates. In contrast, choanal swabs collected from vaccinated birds at 12 wk were 21% and 40% PCR positive for ts-11 strain MG, while those from nonvaccinates were negative. All samples were PCR negative for field strain MG. The pattern of seroconversion as measured by RSPA test in small groups of broiler breeders was different from that previously reported for leghorns. Lateral spread of the ts-11 strain to commingled pen mates occurred rapidly, causing RSPA seroconversion patterns that mimicked those of the vaccinated pen mates.

Dispersion equations for vacuum-deposited tilted-columnar biaxial media.

We consider the application of the Bragg-Pippard (BP) equations for form birefringence to a tilted-columnar biaxial thin film with columns of index n(c) and voids of known index n(v). In such a situation the three forward BP equations that express the principal refractive indices n1, n2, and n3 as functions of n(c), n(v), the packing fraction p(c), and the depolarization factors L1, L2, and L3 can be inverted. The procedure described for adding dispersion to the principal indices involves entry to the BP model via the inverted equations, modification of n(c) to allow for dispersion, and then exit from the model via the forward BP equations. We discuss the introduction of composite columns to the model to allow for angular dependence of n(c) and the selection of suitable dispersion functions for bulk tantalum oxide, titanium oxide, and zirconium oxide. Theory and experiment both show that the dispersion of the normal-incidence birefringence Deltan of the thin films is several times larger than the dispersion of the individual principal refractive indices.

Refined crystal structure of Cd, Zn metallothionein at 2.0 A resolution.

The crystal structure of Cd5,Zn2-metallothionein from rat liver has been refined at 2.0 A resolution of a R-value of 0.176 for all observed data. The five Cd positions in the asymmetric unit of the crystal create a pseudo-centrosymmetric constellation about a crystallographic 2-fold axis. Consequently, the distribution of anomalous differences is almost ideally centrosymmetric. Therefore, the previously reported metal positions and the protein model derived therefrom are incorrect. Direct methods were applied to the protein amplitudes to locate the Cd positions. The new positions were used to calculate a new electron density map based on the Cd anomalous scattering and partial structure to model the metal clusters and the protein. Phases calculated from this model predict the positions of three sites in a (NH4)2WS4 derivative. Single isomorphous replacement phases calculated with these tungsten sites confirm the positions of the Cd sites from the new direct methods calculations. The refined metallothionein structure has a root-mean-square deviation of 0.016 A from ideality of bonds and normal stereochemistry of phi, phi and chi torsion angles. The metallothionein crystal structure is in agreement with the structures for the alpha and beta domains in solution derived by nuclear magnetic resonance methods. The overall chain folds and all metal to cysteine bonds are the same in the two structure determinations. The handedness of a short helix in the alpha-domain (residues 41 to 45) is the same in both structures. The crystal structure provides information concerning the metal cluster geometry and cysteine solvent accessibility and side-chain stereochemistry. Short cysteine peptide sequences repeated in the structure adopt restricted conformations which favor the formation of amide to sulfur hydrogen bonds. The crystal packing reveals intimate association of molecules about the diagonal 2-fold axes and trapped ions of crystallization (modeled as phosphate and sodium). Variation in the chemical and structural environments of the metal sites is in accord with data for metal exchange reactions in metallothioneins.

Speech and language development in preschool twins.

While language, articulation and reading problems have been well documented in young twins, it is not clear how extensive such problems can be or how early in childhood they become evident. At the age of 30 months, twin boys in the La Trobe Twin Study were 8 months behind matched singletons and twin girls on expressive language and 6 months behind on verbal comprehension. They were also 5 months behind on symbolic play and this delay was closely related to language. "Secret" language characterised most of the twin boys but not the girls and the relation of this to articulation delays is discussed. To examine if exposure to other children helps the twin boys, 38-53 month old twins and singletons were matched on the Columbia Mental Maturity Scale at the time of starting preschool. The twin boys had more articulation problems and all twins scored much lower on a Sociability questionnaire completed by the teacher. After 8 months at preschool, all children had advanced in Sociability, but the twins remained just as far behind with poor Sociability relating to poor articulation. The role of intervention programs is discussed.

Measurement of glycosylated haemoglobins and glycosylated plasma proteins in animal models with diabetes or inappropriate hypoglycaemia.

Stable glycosylated haemoglobins and glycosylated plasma proteins were determined by affinity chromatography using Glycogel B in animal models with diabetes or inappropriate hypoglycaemia. Adult Aston ob/ob mice and C57BL/KsJ db/db mice exhibited 1.5-1.9 fold increases of body weight, 2.5-3.4 fold elevations of plasma glucose and 20.9-29.3 fold elevations of plasma insulin. Glycosylated haemoglobins and glycosylated plasma proteins were raised 7.2-8.2 fold and 6.6-6.7 fold respectively. In adult NEDH rats, administration of streptozotocin or implantation of transplantable insulinoma fragments produced reciprocal changes in insulin and glucose concentrations either resulting in the onset of insulin deficiency (5.9 fold decrease) and hyperglycaemia (3.2 fold increase) by 2 days, or hyperinsulinaemia (2.1 fold increase) and hypoglycaemia (1.4 fold decrease) by 6 and 8 days, respectively. Glycosylated plasma proteins were increased (1.2 fold) rapidly after streptozotocin treatment followed by glycosylated haemoglobins (1.6 fold increase) after 8 days. In contrast, the decreases in glycosylated plasma proteins and glycosylated haemoglobins (4.4 fold and 1.4 fold, respectively) in insulinoma-bearing rats preceded the demonstration of hypoglycaemia by 5 and 2 days, respectively. Glycosylated plasma proteins in insulinoma-bearing rats returned to pretransplantation values at 10-16 days. Good correlations were observed in mice and rats between glucose and both glycosylated haemoglobins (r = 0.92 and r = 0.90, respectively) and glycosylated plasma proteins (r = 0.85 and r = 0.93), and between the glycosylated blood proteins themselves (r = 0.95 and r = 0.91). The results show that the measurement of glycosylated blood proteins by affinity chromatography using Glycogel B provides a sensitive and reliable indicator of the recent glycaemic environment.

Lysozyme: a major secretory product of a human colon carcinoma cell line.

One of the major proteins secreted by an established human colon adenocarcinoma cell line has been isolated in 25% yield from the serum-free medium in which the cells were grown and identified as lysozyme. Its purification was achieved by sequential steps of acidification, cation-exchange chromatography, and reversed-phase high-performance liquid chromatography. It was recognized to be a human lysozyme on the basis of its molecular weight (14 000), isoelectric point (10.5), amino acid composition, and enzymatic activity. Its identity with previously characterized human lysozymes was established by amino-terminal sequence, peptide composition, immunological properties, NMR, and crystallography. A 4-day, 7-L collection of conditioned medium contained 20.3 mg of secreted protein of which 4.9 mg or approximately 24% of the total was tumor-derived lysozyme. The intracellular level of lysozyme was approximately 18 ng per 10(6) carcinoma cells. The possible significance of these findings in regard to the malignant process and tumor maintenance is discussed.