Effects of plasmid-mediated growth hormone releasing hormone supplementation in young,healthy Beagle dogs

Our study focused on the evaluation of the pharmacological and toxicological effects of plasmid-mediated GHRH supplementation with electroporation in normal adult dogs over a 180-d period. Twenty-eight dogs (< 2 yr of age) were randomized to four groups. Three groups (four dogs/sex for each group) were treated with ascending doses of GHRH-expressing plasmid: 0.2, 0.6, and 1 mg. One group (two dogs of each sex) served as the control. Clinical observations and body weights were recorded. Hematological, serum biochemical, and urine analyses were performed. Serum IGF-I, ACTH, and insulin were determined. Necropsies were performed on d 93 and 180; organs were weighed and tissues were fixed and processed for light microscopy. Selected tissues were used to assess plasmid biodistribution on d 93. At all doses, plasmid GHRH caused increased weight gain (P < 0.001), without organomegaly. Serum glucose and insulin in fasted dogs remained within normal ranges at all time points. Adrenocorticotropic hormone was normal in all groups. Significant increases in number of red blood cells, hematocrit, and hemoglobin (P < 0.01) were observed. In conclusion, our study shows that plasmid-mediated GHRH supplementation is safe in electroporated doses up to 1.0 mg in young healthy dogs.     

Toxic shock syndrome in a horse with Staphylococcus aureus pneumonia

A 3-year-old Thoroughbred gelding was examined because of clinical signs of pneumonia and shock. Mucous membrane petechiation and ventral edema were observed and considered to be a result of vasculitis. Epidermal necrosis developed on the distal portions of the limbs. The horse had a persistent high fever that was unresponsive to nonsteroidal anti-inflammatory treatment, and Staphylococcus aureus was isolated from a nasal swab specimen and 2 transtracheal wash fluid samples. Antimicrobial, anti-inflammatory, and supportive treatment resulted in clinical improvement. However, resolution of the pulmonary infection required long-term (42 days) antimicrobial administration. Staphylococcus aureus strains isolated from this horse were positive for the toxic shock syndrome toxin-1 gene and were shown to produce toxic shock syndrome toxin-1, the causative factor in toxic shock syndrome in humans. The horse's clinical signs were attributed to toxic shock syndrome secondary to pulmonary S. aureus infection.     

Influence of milk production potential on forage dry matter intake by multiparous and primiparous Brangus females

Brangus cows (n = 29) were used in three experiments to evaluate the effects of parity (multiparous vs. primiparous) and potential genetic merit for milk production (high vs. low) on forage intake during late gestation, early lactation, and late lactation. Cows were selected for milk production based on their sire's EPD for milk production (MEPD). Cows had ad libitum access to (130% of previous 2-d average intake) low-quality hay (5.3% CP and 76% NDF), and cottonseed meal was supplemented to ensure adequate degradable intake protein. All females were adapted to diets for at least 7 d, and individual intake data were collected for 9 d. During the lactation trials, actual milk production was determined using a portable milking machine following a 12-h separation from calves. During late gestation, multiparous cows consumed 24% more (P = 0.01) forage DM (kg/d) than primiparous cows; however, parity class did not influence forage intake when intake was expressed relative to BW. Furthermore, MEPD did not influence forage intake during late gestation. During early lactation, multiparous cows produced 66% more (P < 0.001) milk than primiparous cows, and high MEPD tended (P = 0.10) to produce more milk than low MEPD. Multiparous cows consumed 19% more (P < 0.0001) forage DM than did primiparous cows when expressed on an absolute basis, but not when expressed on a BW basis. High-MEPD cows consumed 8% more (P < 0.05) forage DM than did low-MEPD cows. During late lactation, multiparous cows produced 84% more milk than primiparous cows, although MEPD did not influence (P = 0.40) milk yield. In addition, multiparous cows consumed 17% more (P < 0.01) forage DM per day than primiparous cows, but when intake was expressed relative to BW, neither parity nor MEPD influenced forage DMI during late lactation. Milk yield and BW explained significant proportions of the variation in forage DMI during early and late lactation. Each kilogram increase in milk yield was associated with a 0.33- and 0.37-kg increase in forage DMI for early and late lactation, respectively. Results suggest that multiand primiparous cows consume similar amounts of low-quality forage DM, expressed per unit of BW, during late gestation and lactation. Selecting beef cows for increased genetic merit for milk production increases forage DMI during early lactation.     

Identification of unique or elevated levels of kernel proteins in aflatoxin-resistant maize genotypes through proteome analysis

ABSTRACT Aflatoxins are carcinogens produced by Aspergillus flavus and A. parasiticus during infection of susceptible crops such as maize (Zea mays L.). Resistant maize genotypes have been identified, but the incorporation of resistance into commercial lines has been slow due to the lack of selectable markers. Here we report the identification of potential markers in resistant maize lines using a proteomics approach. Kernel embryo proteins from each of two resistant genotypes have been compared with those from a composite of five susceptible genotypes using large format two-dimensional gel electrophoresis. Through these comparisons, both quantitative and qualitative differences have been identified. Protein spots have been sequenced, and based on peptide sequence homology analysis, are categorized as follows: storage proteins (globulin 1 and globulin 2), late embryogenesis abundant (LEA) proteins related to drought or desiccation (LEA3 and LEA14), water- or osmo-stress related proteins (WSI18 and aldose reductase), and heat-stress related proteins (HSP16.9). Aldose reductase activity measured in resistant and susceptible genotypes before and after infection suggests the importance of constitutive levels of this enzyme to resistance. Results of this study point to a correlation between host resistance and stress tolerance. The putative function of each identified protein is discussed.     

Evaluation of cystatin C as an endogenous marker of glomerular filtration rate in dogs

Cystatin C is a cysteine protease inhibitor produced by all nucleated cells. It is freely filtered by the glomerulus and is unaffected by nonrenal factors such as inflammation and gender. Because of greater sensitivity and specificity, cystatin C has been proposed to replace creatinine as a marker of glomerular filtration rate (GFR) in humans. The aims of this study were to validate an automated assay in canine plasma and to evaluate the usefulness of cystatin C as a marker of GFR in dogs. Western blotting was used to demonstrate cross-reactivity of an anti-human cystatin C antibody. An immunoturbidimetric assay was used to detect cystatin C in 25 clinically healthy dogs and 25 dogs with renal failure. Mean cystatin C concentration in the healthy dogs and the dogs with renal failure was 1.08 +/- 0.16 mg/L and 4.37 +/- 1.79 mg/L respectively. Intra- and interassay variability was <5%. The assay was linear (r = .974) between 0.14 and 7.53 mg/L. Both cystatin C and creatinine concentrations were measured in banked, frozen serum from 20 remnant kidney model dogs and 10 volume-depleted dogs for which GFR measurements by exogenous creatinine clearance had been determined previously. In the remnant kidney model, cystatin C was better correlated with GFR than creatinine (r = .79 versus .54) but was less well correlated with GFR in volume-depleted dogs (r = .54 versus .95). GFR measurements were repeated in the remnant kidney model dogs 60 days after initial GFR measurements. At this time, cystatin C and creatinine concentrations correlated equally well with GFR (r = .891 versus .894, respectively). Cystatin C concentration is a reasonable alternative to creatinine for screening dogs with decreased GFR due to chronic renal failure.     

Use of computerized image analysis to quantify staphylococcal adhesion to canine corneocytes: does breed and body site have any relevance to the pathogenesis of pyoderma?

An optimized system of computerized image analysis was used to investigate variations in the adherence of Staphylococcus intermedius to canine corneocytes from four different breed groups and six different anatomical sites. S. intermedius showed significantly greater adherence to the head and neck compared with the dorsum, but adherence to the limb, axilla and groin did not differ from other sites. Furthermore, there was significantly greater adherence of S. intermedius to corneocytes from the dorsum, forelimb, axilla and groin of Boxers and Bull Terriers than Spaniels and Hounds. S. intermedius, and also Pseudomonas aeruginosa, exhibited abundant adherence, which was significantly greater than Staphylococcus aureus, Streptococcus canis, Klebsiella pneumoniae and Escherichia coli. In addition, S. intermedius adherence demonstrated a sigmoid dose-response curve with increasing bacterial concentration. These results suggest that S. intermedius adheres to canine corneocytes by a specific receptor-ligand interaction and adheres to the skin of some breeds more avidly than others. However, variations in adherence between body regions would not account for the predilection sites of canine bacterial pyoderma.     

Emergence of a New Cucurbit-Infecting Begomovirus Species Capable of Forming Viable Reassortants with Related Viruses in the Squash leaf curl virus Cluster

ABSTRACT Cucurbit leaf curl virus (CuLCV), a whitefly-transmitted geminivirus previously partially characterized from the southwestern United States and northern Mexico, was identified as a distinct bipartite begomovirus species. This virus has near sequence identity with the previously partially characterized Cucurbit leaf crumple virus from California. Experimental and natural host range studies indicated that CuLCV has a relatively broad host range within the family Cucurbitaceae and also infects bean and tobacco. The genome of an Arizona isolate, designated CuLCV-AZ, was cloned and completely sequenced. Cloned CuLCV-AZ DNA A and B components were infectious by biolistic inoculation to pumpkin and progeny virus was transmissible by the whitefly vector, Bemisia tabaci, thereby completing Koch's postulates. CuLCV-AZ DNA A shared highest nucleotide sequence identity with Squash leaf curl virus-R (SLCV-R), SLCV-E, and Bean calico mosaic virus (BCaMV) at 84, 83, and 80%, respectively. The CuLCV DNA B component shared highest nucleotide sequence identity with BCaMV, SLCV-R, and SLCV-E at 71, 70, and 68%, respectively. The cis-acting begomovirus replication specificity element, GGTGTCCTGGTG, in the CuLCV-AZ origin of replication is identical to that of SLCV-R, SLCV-E, and BCaMV, suggesting that reassortants among components of CuLCV-AZ and these begomoviruses may be possible. Reassortment experiments in pumpkin demonstrated that both reassortants of CuLCV-AZ and SLCV-E A and B components were viable. However, for CuLCV-AZ and SLCV-R, only one reassortant (SLCV-R DNA A/CuLCV-AZ DNA B) was viable on pumpkin, even though the cognate component pairs of both viruses infect pumpkin. These results demonstrate that reassortment among sympatric begomovirus species infecting cucurbits are possible, and that, if generated in nature, could result in begomoviruses bearing distinct biological properties.