Soluble scute proteins of healthy and ill desert tortoises (Gopherus agassizii)

OBJECTIVES: To characterize protein composition of shell scute of desert tortoises and to determine whether detectable differences could be used to identify healthy tortoises from tortoises with certain illnesses. ANIMALS: 20 desert tortoises. PROCEDURES: Complete postmortem examinations were performed on all tortoises. Plastron scute proteins were solubilized, scute proteins were separated by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and proteins were analyzed, using densitometry. Two-dimensional immobilized pH gradient-PAGE (2D IPG-PAGE) and immunoblot analysis, using polyclonal antisera to chicken-feather beta keratin and to alligator-scale beta keratin, were conducted on representative samples. The 14-kd proteins were analyzed for amino acid composition. RESULTS: The SDS-PAGE and densitometry revealed 7 distinct bands, each with a mean relative protein concentration of > 1 %, ranging from 8 to 47 kd, and a major protein component of approximately 14 kd that constituted up to 75% of the scute protein. The 2D IPG-PAGE revealed additional distinct 62- and 68-kd protein bands. On immunoblot analysis, the 14-, 32-, and 45-kd proteins reacted with both antisera. The 14-kd proteins had an amino acid composition similar to that of chicken beta keratins. There was a substantial difference in the percentage of the major 14-kd proteins from scute of ill tortoises with normal appearing shells, compared with 14-kd proteins of healthy tortoises. CONCLUSIONS AND CLINICAL RELEVANCE: The major protein components of shell scute of desert tortoises have amino acid composition and antigenic features of beta keratins. Scute protein composition may be altered in tortoises with certain systemic illnesses.     

Molecular and serologic characterization, pathogenicity, and protection studies with infectious bronchitis virus field isolates from California

In this study, we characterized three variant infectious bronchitis virus (IBV) strains isolated in 2003 and 2004 from broiler chickens in California and compared them to previously isolated California variant viruses and to common vaccine serotypes used in the United States. We conducted genetic, serologic, and pathogenicity studies on all three isolates, then tested different vaccines against one of the viruses. Genetically the three variant IBV strains, designated CA557/03, CA706/03, and CA1737/04, were not related to each other. GenBank BLAST database search and phylogenetic analysis of the hypervariable region of the S1 subunit of the spike gene to determine the most closely related viruses to the three variants showed the CA557/03 variant to be 81.8% similar to the CAV/CA56b/91 whereas the CA706/03 and CA1737/04 variant viruses were only distantly related to Dutch/D1466/81 (72.2%), a vaccine strain used in Europe, and Korea/K142/02 (72.7%), a Korean field isolate, respectively. Cross virus-neutralization testing showed that none of the 2003-04 California IBV variant viruses were serologically related to each other or to Ark, Conn, or Mass vaccine strains. In addition the CA1737/04 isolate was also tested against DE072 and found not to be serologically related. All three variant viruses were pathogenic in 1-wk-old broilers and vaccination with Mass/Conn followed by Holland/Conn provided 80% protection against the CA1737/04 virus. The 2003-04 California variant viruses were not compared with variants isolated in California during 1970s and 1980s because, to our knowledge, no genetic information is available and those viruses are no longer obtainable. This study shows that the CA557/03 virus was distantly related to the CAV-type viruses isolated in California in the early 1990s, but that none of the 2003-04 viruses were similar genetically or serologically to the CAL99-type viruses, indicating that new IBV variants continue to emerge and cause disease in commercial chickens in California.     

Exocrine Pancreatic Insufficiency in Dogs and Cats: Online Support for Veterinarians and Owners

Pet owners increasingly refer to online sources before, during, and after seeking veterinary advice for their pets. Although some online information is inaccurate or unhelpful, there is much useful material, and we believe much is to be gained by directing owners to online material that supports and complements veterinary advice. With regard to managing veterinary patients with exocrine pancreatic insufficiency (EPI), there is an excellent online community that not only offers moral support and practical advice, but also facilitates clinical research and fundraising. For genetic diseases like canine EPI that affect primarily young animals, but for which a decade or more of treatment can reasonably be expected, online support can greatly facilitate optimal lifelong management. In this article we provide some information that may be of value when veterinarians consider which web sites to recommend to their clients managing patients with EPI.     

Procedure for processing of central nervous system tissue for immunofluorescence, light, and electron microscopic evaluation.

A rapid and simple procedure has been developed for the simultaneous localization of viral antigen in central nervous system tissues by immunofluorescnece and concomitant correlation with viral induced lesions in the contralateral brain hemisphere. In this study, one-half of the brain was excised from the dog under deep general anesthesia and samples were collected for immunofluorescence by snap freezing in liquid nitrogen. The other half of the brain was perfused under pressure with a buffered glutaraldehyde-paraformaldehyde mixture for subsequent light microscopic and ultrastructural evaluation. The technique was useful in studying central nervous system diseases in large outbred experimental subjects in which economic and other considerations developed for tissue processing in rodents was not feasible.     

Influence of cornual insemination on endometrial damage and microbial flora in the bovine uterus

The objectives were to determine effects of cornual insemination on endometrial characteristics and uterine bacterial flora. Forty cows were assigned randomly (10/group) to each of the following treatments: 1) control--no uterine manipulation; 2) uterine body insemination--deposition of the inseminate at the internal cervical os; 3) cornual insemination--one-half of inseminate deposited into each uterine horn anterior to the internal bifurcation and 4) cornual insemination plus endometrial biopsy--insemination into each uterine horn and trauma induced by endometrial biopsy. Animals were slaughtered and reproductive tracts were removed 5 d after treatment. Bacterial populations of the uterine lumen were evaluated by both flushings and swabbings of each horn. Treatment did not influence the presence of luminal bacteria as determined by flushing or swab samples. Gross endometrial characteristics were quantitated in all tracts. Characteristics considered were total endometrial surface area, discolored regions that appeared to be distinctly darker than the remaining portion of the endometrium and obvious surface damage. Discolored regions were evaluated as both total areas and percentage of endometrium. Treatment did not influence total or percent discolored areas. Microscopic evaluation of these tissues revealed increased edema when compared with nondiscolored regions but no disruption of the luminal epithelium was found. Surface damage was found only in the biopsied treatment group. Observation of the endometrium revealed that extensive regeneration occurred by 5 d after treatment. The data suggest that cornual insemination does not result in uterine trauma or bacterial presence 5 d after breeding, and the endometrium is capable of regeneration after trauma during estrus.     

The effects of undernutrition of suckled pigs on subsequent growth and body composition after nutritional rehabilitation

The effect of nutritional restriction and rehabilitation imposed between 5 and 35 d and 35 and 166 d of age, respectively, on body and tissue growth were investigated in 96 Yorkshire pigs from 12 litters. One-half of the pigs were allowed to suckle continuously until weaning (fully fed, FF); the remainder were removed from the sow for 16 h of each 24 h period (1600 to 0800 h), during which time they received only water (restricted feeding, RF). All pigs were fed ad libitum from 35 d of age. Although significantly lower body and tissue (liver, kidney and gastrocnemius muscle) weights were observed in the RF pigs at 35 and 70 d of age, by 166 d, because of compensatory growth between 35 and 166 d, the difference in body and muscle weights between the two groups of pigs (RF and FF) was reduced to 5%, but the difference in kidney and liver weight remained as high as 15 and 9%, respectively. Total DNA, RNA and protein in the tissues examined were also lower in the RF pigs at 35 and 70 d of age, but increased after ad libitum feeding to more than 90% of the corresponding value in the FF pigs by the time they reached 166 d of age. The results of these measurements and the comparison of the protein: DNA ratios of the tissues indicated that growth retardation in the RF pigs was associated with reduced cell proliferation in liver and kidney and nuclear proliferation in muscle rather than any decrease in cell (or fiber) volume. Percentage of fat was lower while percentage of protein was higher (on a dry matter basis) in the RF pigs than in the FF pigs at 35 d of age. No significant difference in fat and protein content in the carcasses between the RF and FF pigs at 70 or 166 d of age or 90 kg body weight was noted. The carcass of intact males had more protein and less fat than that of females.     

Reliably colonising broiler chickens with Campylobacter spp. using a litter-based method

ABSTRACT

1. Chicken-associated Campylobacter spp. are the cause of most food poisoning cases in Europe. In order to study the host–pathogen interactions, a reliable and reproducible method of colonising chickens with the bacteria is required.

2. This study aimed to identify a more appropriate and less invasive method of colonisation (cf. gavaging) by seeding bedding material (litter) that commercial chickens are kept on with a mixture of Campylobacter spp., broth and faeces.

3. The first phase of the study tested the longevity of Campylobacter spp. recovery in seeded litter over 24 h: significantly more Campylobacter spp. was recovered at 0 or 3 h post-seeding than at 6 and 24 h post-seeding, indicating that the pathogen can survive to detectable levels for at least 3 h in this environment.

4. In the second phase, three groups of 10 broiler chickens (negative for Campylobacter spp. prior to exposure) were exposed at 21 days of age to one of three different Campylobacter jejuni and C. coli mixes (A, B, C), using the method above. At 28 days of age, birds were euthanised by overdose of barbiturate or cervical dislocation, and livers and caeca removed for Campylobacter spp. assessment.

5. All liver and 28/30 caeca samples tested positive for Campylobacter spp., with mix A and C giving higher counts in the caeca than mix B. The method of euthanasia did not affect Campylobacter spp. counts.

6. In conclusion, a successful method for reliably colonising broiler chickens with Campylobacter spp. has been developed which negates the need for gavaging and is more representative of how contamination occurs in the field.