Genomic sequence and cardiac expression of atrial natriuretic peptide in cats

OBJECTIVE: To determine the nucleotide and amino acid sequence of atrial natriuretic peptide (ANP) in cats and its typical regions of cardiac expression. ANIMALS: 5 healthy adult mixed-breed cats. PROCEDURE: Total RNA was extracted from samples obtained from the left and right atrium, left and right ventricle, and interventricular septum of each cat. The RNA was used to produce cDNA for sequencing and northern blot analysis. Genomic DNA was extracted from feline blood samples. Polymerase chain reaction primers designed from consensus sequences of other species were used to clone and sequence the feline ANP gene. RESULTS: The feline ANP gene consists of 1,072 nucleotides. It consists of 3 exons (123, 327, and 12 nucleotides) separated by 2 introns (101 and 509 nucleotides). It has several typical features of eukaryotic genes and a putative steroid-response element located within the second intron. Preprohormone ANP consists of 153 amino acids. The amino acid sequence of the active form of feline ANP (ANP-30) is identical to that of equine, bovine, and ovine ANP-30 and differs from that of human, canine, and porcine ANP-28 only by 2 carboxy-terminal arginine residues. The ANP mRNA was detected only in the left and right atria. CONCLUSIONS AND CLINICAL RELEVANCE: The genetic and protein structure and principal regions of cardiac expression of feline ANP are similar to those of other species. Results of this study should be helpful in future studies on the natriuretic response in cats to diseases that affect cardiovascular function.     

Multiple-center study of reduced-concentration triamcinolone topical solution for the treatment of dogs with known or suspected allergic pruritus

OBJECTIVE: To determine the efficacy of triamcinolone acetonide topical solution (TTS) in dogs for use in reduction of clinical signs of pruritic inflammatory skin diseases of a known or suspected allergic basis and to evaluate adverse effects associated with TTS administration. ANIMALS: 103 pruritic adult dogs with known or suspected allergic skin disease. PROCEDURE: Dogs were treated for 4 weeks with TTS or with vehicle solution (control dogs) in a multiple-center study. Clinical signs were scored by owners and by examining veterinarians before and after treatment. Blood samples obtained before and after treatment were subjected to routine hematologic and serum biochemical analyses. RESULTS: Treatment success, as defined by an improvement of at least 2 of 6 grades in overall clinical score, was evident in 35 of 52 (67%) TTS-treated dogs (mean improvement, 1.98) and 12 of 51 (24%) control dogs (mean improvement, 0.29). For several criteria, TTS was significantly more effective than vehicle in reducing clinical signs. Minor alterations in hematologic determinations in TTS-treated dogs were limited to slightly lower total leukocyte, lymphocyte, and eosinophil counts after treatment. Minor adverse effects were reported by owners in 6 of 52 (12%) TTS-treated and 9 of 51 (18%) control dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Triamcinolone used as a spray solution at a concentration approximately one-sixth the concentration of triamcinolone topical preparations currently available for veterinary use is effective for short-term alleviation of allergic pruritus in dogs. Adverse effects are few and mild and, thus, do not preclude prolonged treatment with the solution.     

Cloning and sequence analysis of the complementary DNA for feline preproparathyroid hormone

OBJECTIVES: To clone and sequence the cDNA for feline preproparathyroid hormone (preproPTH) and to compare that sequence with other known parathyroid hormone (PTH) sequences. SAMPLE POPULATION: Parathyroid glands from 1 healthy cat. PROCEDURES: A cDNA library was constructed in lambda phage from feline parathyroid gland mRNA and screened with a radiolabeled canine PTH probe. Positive clones were sequenced, and nucleic acid and deduced amino acid sequences were analyzed and compared with known preproPTH and PTH sequences. RESULTS: Screening of approximately 2 X 10(5) recombinant plaques revealed 3 that hybridized with the canine PTH probe; 2 clones comprised the complete sequence for feline preproPTH. Feline preproPTH cDNA consisted of a 63-base pair (bp) 5'-untranslated region (UTR), a 348-bp coding region, and a 326-bp 3'-UTR. The coding region encoded a 115-amino acid peptide. Mature feline PTH consisted of 84 amino acids. Amino acid sequence analysis revealed that feline PTH was > 83% identical to canine, bovine, swine, equine, human, and macaque PTH and 69, 71, and 44% identical to mouse, rat, and chicken PTH, respectively. Within the region responsible for hormonal activity (amino acids 1 to 34), feline PTH was > 79% identical to other mammalian PTH sequences and 64% identical to the chicken sequence. CONCLUSIONS AND CLINICAL RELEVANCE: The amino acid sequence of PTH is conserved among mammalian species. Knowledge of the cDNA sequence for feline PTH may be useful to investigate disturbances of calcium metabolism and alterations in PTH expression in cats.     

Udder cleft dermatitis and sarcoptic mange in a dairy herd

OBJECTIVE: To determine prevalence of udder cleft dermatitis in a dairy herd that was experiencing an outbreak of sarcoptic mange. DESIGN: Clinical survey. ANIMALS: 1,597 Holstein cows and late-gestation heifers. PROCEDURE: Animals were examined for udder cleft dermatitis and for skin lesions consistent with sarcoptic or chorioptic mange. Skin scrapings were collected from 56 cows and examined for ectoparasites. The herd was revisited 1 year later, and prevalences of udder cleft dermatitis and lesions consistent with mange were determined in 506 cows. RESULTS: Of the 1,597 cattle examined, 280 (18%) had udder cleft dermatitis, and 1,397 (87.5%) had lesions consistent with mange. In 43 of 56 (77%) cows, skin scrapings revealed Sarcoptes mites. Udder cleft dermatitis was significantly more common in older than in younger cows. In first-lactation cows, udder cleft dermatitis was less common during the first 4 months of lactation than in the later stages of lactation, but udder cleft dermatitis was identified in cows in all stages of lactation and in cows that were not lactating. The herd was treated with eprinomectin to control mites, and prevalence of lesions consistent with mange 1 year later was only 2.8%. However, prevalence of udder cleft dermatitis was still 12%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that cows in any stage of lactation and cows that are not lactating can have udder cleft dermatitis but that lesions are more common in older cows. Control of sarcoptic mange was accompanied by a moderate reduction in the prevalence of udder cleft dermatitis but did not eliminate the condition.     

Vertical position of the patella in the stifle joint of clinically normal large-breed dogs

OBJECTIVE: To define the vertical position of the patella in clinically normal large-breed dogs. SAMPLE POPULATION: Cadavers of 13 clinically normal large-breed dog. PROCEDURE: Both hind limbs were harvested with intact stifle joints and mounted on a positioning device that allowed full range of motion of the stifle joint. Lateral radiographic views were obtained with the stifle joints positioned at each of 5 angles (148 degrees, 130 degrees, 113 degrees, 96 degrees, and 75 degrees). Vertical position of the patella through a range of motion was depicted on a graph of mean stifle angle versus corresponding mean proximal patellar position (PPP) and distal patellar position (DPP) relative to the femoral trochlea for each dog. Ratio of length of the patellar ligament to length of the patella (L:P) was determined for each dog. Overall mean, SD, and 95% confidence intervals for L:P were calculated for all dogs. RESULTS: Evaluation of vertical position of the patella through a range of motion revealed a nearly linear relationship between joint angle and PPP and joint angle and DPPF Evaluation of L:P results did not reveal significant differences between limbs (left or right) or among joint angles. Overall mean +/- SD L:P for all dogs was 1.68 +/- 0.18 (95% confidence interval, 1.33 to 2.03). CONCLUSIONS AND CLINICAL RELEVANCE: The L:P proved to be a repeatable measurement of vertical patellar position, which is independent of stifle angles from 75 degrees to 148 degrees.This measurement could be used as a quantitative method for diagnosing patella alta and patella baja in large-breed dogs.     

Population-based study of fecal shedding of Clostridium perfringens in broodmares and foals

OBJECTIVE: To determine the percentage of broodmares and foals that shed Clostridium perfringens in their feces and classify the genotypes of those isolates. DESIGN: Prospective cross-sectional study. ANIMALS: 128 broodmares and their foals on 6 equine premises. PROCEDURES: Anaerobic and aerobic bacteriologic cultures were performed on feces collected 3 times from broodmares and foals. All isolates of C. perfringens were genotyped. RESULTS: Clostridium perfringens was isolated from the feces of 90% of 3-day-old foals and 64% of foals at 8 to 12 hours of age. A lower percentage of broodmares and 1- to 2-month-old foals shed C. perfringens in their feces, compared with neonatal foals. Among samples with positive results, C. perfringens type A was the most common genotype identified (85%); C. perfringens type A with the beta2 toxin gene was identified in 12% of samples, C. perfringens type A with the enterotoxin gene was identified in 2.1% of samples, and C. perfringens type C was identified in < 1% of samples. CONCLUSIONS AND CLINICAL RELEVANCE: Clostridium perfringens was identified from the feces of all but 6 foals by 3 days of age and is likely part of the normal microflora of neonatal foals. Most isolates from broodmares and foals are C. perfringens type A; thus, the clinical relevance of culture results alone is questionable. Clostridium perfringens type C, which has been associated with neonatal enterocolitis, is rarely found in the feces of horses.     

Genetic and management factors that influence the susceptibility of cattle to Mycobacterium bovis infection

Genetic variation in the susceptibility of cattle to Mycobacterium bovis infection exists in differences between families and species, but not breeds. Susceptibility to M. bovis infection increases with age of cattle. Natural exposure to M. bovis or environmental mycobacteria may assist in the development of specific immunity, but there is no direct evidence for such immunological priming of tuberculosis resistance in cattle. This has, however, been demonstrated in humans and other animals. Since non-specific mechanisms have a role in protective immunity, developing an effective vaccine will be difficult, even though some protection of other species has been achieved. Immunological suppression in the periparturient period can produce anergic reactors, which may act as a constant source of infection for cattle-to-cattle transmission. Circumstantial evidence suggests that an adequate intake of mineral, vitamin and protein reduces the susceptibility of cattle. Although weather patterns have been implicated in the susceptibility of herds to M. bovis infection, there is insufficient information to determine the risk factors precisely. It is concluded that some reduction in the susceptibility of cattle to M. bovis infection can be achieved by modifications to the management system to minimize risk factors, but that a considerable amount of further research is required.     

Development of an assay to determine single nucleotide polymorphisms in the prion gene for the genetic diagnosis of relative susceptibility to classical scrapie in sheep.

The objective of this study was to develop a reliable Taqman 5' Nuclease Assay for genotyping sheep for scrapie susceptibility. The sheep prion gene contains 2 single nucleotide polymorphisms (SNPs) that may mediate resistance to classical scrapie, one at codon 136, alanine (A) or valine (V), and another at codon 171, arginine (R) or glutamine (Q). The R allele appears to confer resistance to classical scrapie, with the AA(136) RR(171) genotype the most resistant to scrapie and QR(171) only rarely infected in the US sheep population. The Assays by Design protocol was used for development of probes and primers for codon 136 and Primer Express for codon 171. Commercially available kits were used to isolate genomic DNA from blood or muscle. For validation, 70 SNP determinations for each codon were compared to commercial testing with an error rate of less than 1%. Then, 935 samples from blood (n = 818) and muscle (n = 117) were tested for both codons with 928 successful determinations and only 7 samples (<1% of total samples) that needed repeating. Genotypes were AA QQ (n = 102; 11.0%), AV QQ (n = 28; 3.0%), AA QR (n = 396; 42.7%), AV QR (n = 54; 5.8%), and AA RR (n = 348; 37.5%). Thus, 86% of the sheep tested (n = 798) contained R at codon 171 and were expected to be scrapie-resistant. This new Taqman 5' Nuclease SNP genotyping assay is accurate, easy to perform, and useful in the study of classical scrapie in sheep and its prevention through selective breeding programs to eliminate highly susceptible animals.