In vitro effects of plant and mushroom extracts on immunological function of chicken lymphocytes and macrophages

1.?The present study was conducted to examine the effects of organic extracts from milk thistle (Silybum marianum), turmeric (Curcuma longa), reishi mushroom (Ganoderma lucidum), and shiitake mushroom (Lentinus edodes) on innate immunity and tumor cell viability.

2.?Innate immunity was measured by lymphocyte proliferation and nitric oxide production by macrophages, and the inhibitory effect on tumor cell growth was assessed using a non-radioactive assay. For measuring the cytokine levels in the HD11 macrophages which were treated with extracts of turmeric or shiitake mushroom, the levels of mRNAs for interferon-α (IFN- α), interleukin-1β (IL-1β), IL-6, IL-12, IL-15, IL-18, and tumor necrosis factor superfamily 15 (TNFSF15) were quantified by real time RT-PCR.

3.?In vitro culture of chicken spleen lymphocytes with extracts of milk thistle, turmeric, and shiitake and reishi mushrooms induced significantly higher cell proliferation compared with the untreated control cells. Stimulation of macrophages with extracts of milk thistle and shiitake and reishi mushrooms, but not turmeric, resulted in robust nitric oxide production to levels that were similar with those induced by recombinant chicken interferon-γ. All extracts uniformly inhibited the growth of chicken tumor cells in vitro at the concentration of 6·3 through 100 µg/ml. Finally, the levels of mRNAs encoding IL-1β, IL-6, IL-12, IL-18, and TNFSF15 were enhanced in macrophages that were treated with extracts of turmeric or shiitake mushroom compared with the untreated control.

4.?These results document the immunologically-based enhancement of innate immunity in chickens by extracts of plants and mushrooms with known medicinal properties in vitro. In vivo studies are being planned to delineate the cellular and molecular mechanisms responsible for their mechanism of action.     

Apocynin activity in spontaneously hypertensive rats (SHR): preliminary studies in vivo

An elevation in angiotensin II (Ang II) levels is a common occurrence in spontaneously hypertensive rats (SHRs). Infusions of Ang II and a high salt diet increase the activity of NADPH oxidase that stimulates superoxide anion (O⁻²) generation and increases the expression of certain subunits of NADPH oxidase. Apocynin, an NADPH oxidase inhibitor with antihypertensive effects, is able to inhibit the release of superoxide anion by inhibiting NADPH oxidase activity and blocking the migration of p47 phox to the mitochondrial membrane. The aim of our study was to evaluate the antihypertensive effects of apocynin in SHRs and Wistar rats (WKYs) using a micropuncture technique. After microperfusion of both the proximal and distal tubules, we found that SHRs treated with apocynin showed a decrease in the free-flow collection of the proximal tubule (PT), which was not affected in WKYs. Moreover, significant differences were not demonstrated in the distal tubule (DT), probably due a mechanism of compensation that occurs in the loop of Henle. In conclusion, it is possible that the mechanisms of reabsorption in the PT are controlled by the interactions of O⁻² and nitric oxide (NO). These data could suggest a higher activity of NADPH oxidase and increase in reactive oxygen species (ROS) production in the PT during hypertension.

     

Genetic parameters for abnormal sucking traits in Austrian Fleckvieh heifers

Cross‐sucking and intersucking are considered abnormal behaviours in cattle and constitute a common problem in dairy farming. Cross‐sucking in calves is defined as sucking any body parts of another calf whereas intersucking in heifers and cows is defined as sucking the udder or udder area. The aim of this study was to determine the genetic variability for abnormal sucking behaviour by estimating genetic parameters and examining individual differences between sires with large progeny groups. By means of a questionnaire, cattle breeders in the federal state Lower Austria were requested to identify all currently kept animals which are known of either inter‐ or cross‐sucking (both defined as the same binary trait ‘sucking’ with 0 and 1 referring to the absence and presence of this abnormal behaviour) or allowing sucking (also treated as a binary trait, scored as 1 if an animal was known of allowing herd mates to suck and 0 otherwise). Records of 1222 farms and 13 332 dual purpose Simmental females aged between 21 and 700 days were investigated applying a linear animal model with fixed herd × year × season and random genetic animal effect and a threshold sire model with the herd × year × season effect being treated as random. In total, 8.6% and 4.1% of all calves/heifers were observed sucking and allowing sucking, respectively. Heritabilities of 0.040 ± 0.014 and 0.007 ± 0.006 (linear animal model) and 0.116 ± 0.041 and 0.026 ± 0.024 (threshold model) were found for the traits sucking and allowing sucking, respectively. Breeding values were estimated applying the same models for the trait sucking. Taking all 254 sires into account, the Pearson and Spearman correlation coefficients between breeding values estimated by linear animal and sire threshold model were 0.86 and 0.80. Thus, little difference was observed between the two methods.     

Separation of protein and lactose intake over meals dissociates postprandial glucose and insulin concentrations and reduces postprandial insulin responses in heavy veal calves

The present study examined, at identical daily nutrient intakes, the impact of separating protein and lactose intakes across two daily meals on the metabolic and endocrine status in heavy veal calves. Calves were assigned to one of six degrees of separating protein and lactose over the two meals (termed nutrient synchrony, SYN 1-6; 6 calves/treatment). They were fed the protein-rich (P-)meal and the lactose-rich (L-)meal at 06:00 and 18:00h, respectively, or vice versa. At SYN 1, calves were fed with 50% of the daily protein and 50% of the daily lactose intake in each meal. Protein and lactose were iso-energetically exchanged between the two daily meals from SYN 1 to 6. At SYN 6, 85% of the daily protein and 20% of the daily lactose was fed in the P-meal and the remainder in the L-meal. Blood samples were collected hourly during 24h. Mean 24h glucose concentrations increased and insulin concentrations decreased from SYN 1 to 6. Postprandial 5h areas under concentration curves (AUC(0-5h)) of glucose increased with increasing meal lactose content. AUC(0-5h) of non-esterified fatty acids increased after P- and L-meals from SYN 1 to 6. Urea concentrations increased after L-meals from SYN 1 to 6, but decreased after P-meals from SYN 1 to 6. Insulin AUC(0-5h) decreased after L-meals and after P-meals from SYN 1 to 6. Nutrient asynchrony did not affect insulin-like growth factor-1, glucagon, growth hormone, leptin, 3,5,3'-triiodothyronine and thyroxine. In conclusion, separation of protein and lactose intake over meals inhibited insulin responses to a lactose-rich meal in heavy veal calves despite high plasma glucose concentrations.     

Canine lymphoproliferative disease characterized by lymphocytosis: immunophenotypic markers of prognosis

BACKGROUND: Canine lymphoproliferative disease often presents with lymphocytosis and is immunophenotypically diverse. HYPOTHESIS: Immunophenotype predicts prognosis in canine lymphoproliferative disorders involving circulating lymphocytosis. ANIMALS: Dogs that had peripheral blood evaluation performed by flow cytometry by the Clinical Immunology Service at Colorado State University between 2003 and 2005. METHODS: Outcome data regarding treatment and survival were sought on patients with lymphocytosis comprising a single lymphocyte subset. Ninety-six patients that met the inclusion criteria had sufficient follow-up information to be included in the study. RESULTS: Four main phenotypic classifications were found: CD8+ T-cell, CD21+ B-cell, CD4-8-5+ (aberrant T-cell phenotype), and CD34+ (undifferentiated progenitor). Expression of CD34 predicted poor outcome with median survival of 16 days (P < .0001) compared with other phenotypes. Within the CD8+ phenotype, dogs presenting with a lymphocytosis >30,000 lymphocytes/muL had significantly shorter median survival (131 days) than those presenting with <30,000 lymphocytes/muL (1098 days, P < .0008). Within the T-cell leukemias, there was no difference in outcome between dogs with CD4-8-5+ leukemia and dogs with the CD8+ T-cell phenotype nor was the loss of expression of the pan-leukocyte marker CD45 associated with decreased survival time. A CD21+ lymphocytosis composed of large cells was associated with shorter survival time (129 days) than those with smaller circulating cells (median survival not reached, P < .01). CONCLUSIONS AND CLINICAL IMPORTANCE: Immunophenotyping provides an objective method for determining prognosis in lymphoproliferative disorders characterized by lymphocytosis.     

Postoperative bleeding in retired racing greyhounds

BACKGROUND: Some retired racing Greyhounds (RRG) that undergo surgery bleed excessively. Hypothesis: Greyhounds that bleed excessively will have one or more preoperative hemostatic abnormalities that can be used to predict the risk and severity of postoperative bleeding. ANIMALS: Eighty-eight RRG undergoing ovariohysterectomy or castration. METHODS: All dogs were evaluated preoperatively with a physical exam, CBC, platelet count, OSPT, APTT, platelet function with PFA-100(a); fibrinogen, d-dimer, plasminogen (Plmg), antiplasmin (AP), antithrombin (AT), and vWF concentration (vWF:Ag); vWF collagen binding assay (vWF:CBA), and Factor XIII assay. Assays were repeated in the dogs that bled, and in an age- and sex-matched control group of RRG. RESULTS: Twenty-six percent of the dogs had bleeding 36-48 hours after surgery. AP (P <.0001) and AT concentration (P= .007) were significantly lower, and vWF:CBA (P= .0284) was higher preoperatively in the dogs with excessive hemorrhage. A lower platelet count (P= .001) and hematocrit (P= .002), shorter OSPT (P= .0002) and higher plasma fibrinogen (P <.0001), and AP (P= .001) concentration were detected at the time of bleeding compared with preoperative values in the dogs that bleed excessively. The same findings were observed postoperatively for the control group, except for the decrease in hematocrit. CONCLUSIONS AND CLINICAL IMPORTANCE: The results indicate that this excessive postoperative bleeding is not attributable to a primary or secondary hemostatic defect, but could result from altered fibrinolysis.     

Comparative fluke burden and pathology in condemned and non-condemned cattle livers from selected abattoirs in Zambia

After dissecting 70 condemned and 32 non-condemned cattle livers collected from Lusaka, Chisamba, Mongu and Senanga abattoirs and Turnpike slaughter slab, significantly higher numbers of liver flukes (Fasciola gigantica) (P < 0.001) were found in the condemned livers (mean +/- SD = 100.6 +/- 16.7) than in the non-condemned livers (mean +/- SD = 0.7 +/- 0.5). Liver flukes found in 9.4% of the non-condemned livers suggest that abattoir records of liver inspection may underestimate F. gigantica infections. Average faecal fluke egg counts from animals with condemned livers (5 eggs per gram [EPG]) were significantly higher (P < 0.001) than in animals with non-condemned livers (0.8 EPG). No correlation was found between egg counts and number of flukes. Fibrosis and calcification were common in condemned livers, being severest in the vicinity of the bile ducts. Only two (6.3%) of the non-condemned livers showed pathological changes on the liver edges. The severe liver damage and high worm burden may explain low production levels experienced in cattle in Zambia maintained under traditional systems of management where worm control and good management programmes are rarely practiced.     

Clinical evaluation of a peptide-ELISA based upon N-terminal B-cell epitope of the VapA protein for diagnosis of Rhodococcus equi pneumonia in foals

A total of 227 field samples from naturally exposed foals aged between 3 weeks and 6 months were used in an evaluation of a peptide-based enzyme-linked immunosorbent assay (ELISA) for diagnosis of Rhodococcus equi infection. A biotinylated peptide derived from the virulence-associated protein A (VapA) of R. equi, a horse pathogen, was synthesized and designated as PN11-14. The peptide corresponds to the N-terminal B-cell epitope TSLNLQKDEPNGRASDTAGQ of the VapA protein. Based upon a serum immunoglobulin (Ig)G titre of 512 as a positive cut-off value for the R. equi infection, the ELISA provided the overall sensitivity of 47.62%, specificity of 69.67% and an accuracy of 59.47% with a positive predictive value of 57.47% for true R. equi pneumonia. The assay was improved by detecting VapA-specific IgGb antibodies against N-terminal B-cell epitope of the VapA protein rather than IgG antibodies. The VapA-IgGb ELISA showed the overall sensitivity of 70.47%, specificity of 72.13% and accuracy of 71.36% with a positive predictive value of 68.52%. Diagnosis of R. equi disease in 6-week-old foals showed that the VapA-IgGb ELISA provided an increasing trend (P=0.0572) in sensitivity of 82.4% in comparison with the VapA-IgG ELISA which showed the sensitivity of 58.8%. However, differences in specificity of both tests were statistically insignificant (P=0.357) as analysed by the McNemar test. These results indicated that detection of VapA-specific IgGb antibodies may be a better predictor of R. equi disease in foals.