Estrogen and progesterone receptor status of mammary carcinomas and correlation with clinical outcome in dogs.

Estrogen and progesterone receptors (ER, PR) were measured in cytosol fractions from 18 primary canine mammary carcinomas by use of biochemical assays. One or both receptors were detected (> 10 fmol/mg of cytosol protein) in 11 tumors: 5 ER and PR; 2 ER only; 4 PR only. Mean cytoplasmic receptor concentrations (fmol/mg of cytosol protein) were 22.8 +/- 2.9 (SEM) for ER and 51.0 +/- 10.3 for PR in tumors containing ER and PR, 28.8 +/- 12.1 for ER in tumors containing only ER and 13.2 +/- 1.5 for PR in tumors containing only PR. Estrogen or progesterone receptors or both were identified in 6 of 9 tubular adenocarcinomas, 4 of 5 papillary adenocarcinomas, and 1 of 1 squamous cell carcinoma. These receptors were not identified in solid carcinomas (n = 2) or a single spindle cell carcinoma. Although the number of cases was limited, survival times of dogs tended to be longest in those with tumors containing ER alone or in combination with PR, intermediate in those with tumors containing only PR, and shortest in those with tumors without ER or PR. A correlation was not apparent between receptor status and age, presence of ovaries, tumor size, or histologic classification of the tumor. In the analysis of this series, the extent of surgery (mastectomy of the involved gland vs unilateral or bilateral mastectomy) did not appear to influence the outcome of the disease, and metastasis to regional lymph nodes did not appear to be a reliable prognostic indicator.     

Obesity and diet affect glucose dynamics and insulin sensitivity in Thoroughbred geldings

Insulin resistance is considered a risk factor in obesity, laminitis, exertional rhabdomyolysis, and osteochondrosis. The objective was to use the minimal model to estimate glucose effectiveness (Sg) and insulin sensitivity (Si) in nonobese to obese horses initially adapted to forage only, then adapted to forage plus supplements rich in starch and sugar (SS) or fiber and fat (FF). Ten Thoroughbred geldings, with BCS of 5 (nonobese), 6 (moderately obese), and 7 to 8 (obese), were adapted to pasture and hay, allocated to two groups, and fed SS or FF in a switch-back design with 8 wk of adaptation. Modified frequent-sampling i.v. glucose tolerance tests were applied after adaptation to forage, SS, and FF. For the tolerance tests, horses were kept in stalls overnight and provided hay, and venous catheters were placed the next morning. Baseline samples were collected, 0.3 g of glucose/kg of BW was given i.v., and blood was sampled at 1, 2, 3, 4, 5, 6, 7, 8, 10, 12, 14, 16, and 19 min. At 20 min, 30 mU of insulin/kg of BW was given, followed by sampling at 22, 23, 24, 25, 27, 30, 35, 40, 50, 60, 70, 80, 90, 100, 120, 150, and 180 min. Plasma was analyzed for glucose and insulin, and Si, Sg, acute insulin response to glucose, and the disposition index were calculated. Normality was tested using the Shapiro-Wilk statistic. Body condition effects were analyzed using a mixed model with repeated measures. Diet effects were analyzed using a Wilcoxon signed rank test. The Sg was higher in obese than nonobese (P = 0.003) and moderately obese (P = 0.007) horses; Si was lower in obese than nonobese (P = 0.008) horses, and acute insulin response to glucose was higher in obese than nonobese (P = 0.039) horses. Effects of diet were likely confounded by body condition, but horses had lower Si (P = 0.066) when fed SS compared with FF, especially when nonobese. In conclusion, the minimal model effectively estimated Sg, Si, acute insulin response to glucose, and disposition index in horses. Obese geldings were insulin-resistant and seemed to rely primarily on Sg for glucose disposal. Feeding a diet rich in sugar and starch decreased insulin sensitivity of horses. Maintenance of body condition and avoidance of grain-based meals rich in sugar and starch would be beneficial to decrease the risk of developing insulin resistance and associated metabolic syndromes in horses, especially for horses at risk for these syndromes.     

Polyclonal activation of B cells occurs in lymphoid organs from porcine reproductive and respiratory syndrome virus (PRRSV)-infected pigs

Porcine reproductive and respiratory syndrome virus (PRRSV) induces a persistent viral infection associated with an inefficient humoral immune response. A study of lymphoid B cells and specific humoral immune response was performed in blood and several lymphoid organs collected from PRRSV experimentally-infected pigs. Groups of specific pathogen-free (SPF) pigs were infected with the LHVA-93-3 isolate of PRRSV, and blood, tonsils, spleen and mediastinal lymph nodes (MLN) were collected at various times postinfection (p.i.) (3-60 days). Lymphoid cells were isolated, immunolabeled for cytofluorometric determination of B cell percentages, used for counting specific anti-PRRSV antibody secreting B cells by an ELISPOT assay, or cultured for metabolic activity. The presence of anti-PRRSV antibodies in the serum of infected pigs was determined using a commercial ELISA assay. Virus detection was performed in all tissues, including lungs, by virus isolation and RT-PCR. The results show that percentages of B cells increased in tonsils as soon as 3 days until 17 days p.i. in PRRSV-infected pigs while they increased in spleen at 3 days p.i. only, due to an increase of larger Ig(high)-producing B cells. Metabolic activity of lymphoid cells from blood and spleen increased at 3 days p.i. only while lymphoid cells from tonsils and MLN transiently decreased at that time and increased thereafter up to 60 days p.i. Anti-PRRSV antibody-secreting B cells occurred in tonsils after 10 days p.i. and strongly increased up to 60 days p.i. However, specific anti-PRRSV-secreting B cells were detected in blood and spleen after 17 days p.i and in MLN only after 45 days p.i. Specific antibodies were detectable in serum at 10 days p.i., reached the maximum level at 45 days and remained high up to 60 days p.i. Infectious virus was detected in lungs and MLN as soon as 3 days p.i., and remained detectable up to 45 days p.i. in tonsils of one pig while viral RNA was detected in most organs up to 60 days p.i. In vitro experiments revealed that inactivated virus induced a stimulation of lymphoid cells isolated from PRRSV-infected pigs while it was cytotoxic for lymphoid cells from control pigs. Taken together, these results indicate that viral infection induced simultaneously a polyclonal activation of B cells, mainly in tonsils, and an exaggerated and prolonged specific humoral immune response due to persistent viral infection in lymphoid organs.     

Distribution of immune cells in the female reproductive tract in uninfected and FIV infected cats

Cell-free and cell-associated FIV effectively cross the mucosa of the feline female reproductive tract. To identify possible cellular targets of FIV and to characterize changes in mucosal immunity after infection, we examined the types and numbers of immune cells residing in the reproductive tracts of control and intravaginally FIV-infected cats. Sections of the vestibule, vagina, cervix, uterus, and ovaries, were examined by immunohistochemistry for CD4+ and CD8+ T lymphocytes, CD22+ B lymphocytes, CD1a+ dendritic cells, and CD14+ macrophages. The reproductive tract of uninfected cats contained substantial numbers of CD8+ T lymphocytes, CD4+ T lymphocytes and macrophages, as well as moderate numbers of CD1a+ dendritic cells, and few B lymphocytes. The most prominent change between FIV- and FIV+ cats was a marked decrease in the concentration of CD4+ T lymphocytes resulting in inverted CD4+:CD8+ ratios throughout the reproductive tract of infected cats. There was also a trend towards increasing numbers of CD1a+ dendritic cells in the intravaginally-infected FIV+ cats, and decreasing numbers of macrophages and CD22+ B lymphocytes. This study indicates that similar to the peripheral immune system, FIV infection is associated with CD4+ cell loss and reduced CD4+:CD8+ ratios in the female reproductive mucosal tissue.     

Equine Granulocytic Anaplasmosis: A Case Report and Review

A 15 year-old Thoroughbred mare was examined for lethargy, fever, and inappetence of 1-day duration. A hard-bodied tick was removed from the horse. A complete blood count (CBC) demonstrated leukopenia with lymphopenia and thrombocytopenia. Morulae were visualized in circulating granulocytes. A polymerase chain reaction (PCR) confirmed the identity of these organisms as Anaplasma phagocytophilum. The horse was treated symptomatically for fever and inappetence with flunixin meglumine (1.1 mg/kg [0.5 mg/lb]) and oral electrolyte paste. Oxytetracycline (6.6 mg/kg [3 mg/lb] intravenously every 24 hours) treatment was begun as soon as a definitive diagnosis was determined. The mare responded to treatment, but she was switched to oral doxycycline (10 mg/kg [4.5 mg/lb] every 12 hours) after 5 days because of perivascular swelling at the injection site. Complete resolution of clinical signs was seen. There was no evidence of recurrence 1 year later. No additional horses at the farm were affected. The horse in this report presented for lethargy, inappetence, and fever, with limited other abnormalities. This represents a classical presentation of a mild to moderate case of anaplasmosis, which had not previously been reported in Virginia. The disease may be more widespread than has been previously reported, and it should warrant inclusion on a complete differential diagnosis list in a case of fever of unknown origin.     

In vitro antimicrobial sensitivity of Mycoplasmas isolated from the bovine genital tract.

The in vitro antimicrobial sensitivity of 14 mycoplasma and 13 ureaplasma strains isolated from the genital tracts of bulls was examined. It was found that at relatively low concentrations, tetracycline, declomycin and tylosin were lethal to both types of organisms. Lincospectin, berenil, streptomycin and erythromycin were lethal to mycoplasmas but were only inhibitory to the ureaplasma strains at the same concentrations. Polymyxin B and novobiocin were ineffective at the levels tested.     

Ralstonia solanacearum virulence in tomato seedlings inoculated by leaf clipping

Ralstonia solanacearum is a phytopathogenic bacterium that colonizes the xylem vessels of host plants leading to a lethal wilt disease. Although several studies have investigated the virulence of R. solanacearum on adult host plants, infection studies of this pathogen on the seedling stages of hosts are less common. In a preliminary observation, inoculation of R. solanacearum F1C1 on 6‐ to 7‐day‐old tomato seedlings by a simple leaf‐clip strategy resulted in a lethal pathogenic condition in seedlings that eventually killed these seedlings within a week post‐inoculation. This prompted testing of the effect of this inoculation technique in seedlings from different cultivars of tomato and similar results were obtained. Colonization and spread of the bacteria throughout the infected seedlings was demonstrated using gus‐tagged R. solanacearum F1C1. The same method of inoculating tomato seedlings was used with R. solanacearum GMI1000 and independent mutants of R. solanacearum GMI1000, deficient in the virulence genes hrpB, hrpG, phcA and gspD. Wildtype R. solanacearum GMI1000 was found to be virulent on tomato seedlings, whereas the mutants were found to be non‐virulent. This leaf‐clip technique, for inoculation of tomato seedlings, has the potential to be a valuable approach, saving time, space, labour and costs.