In vitro dose–response of carvacrol, thymol, eugenol and trans-cinnamaldehyde and interaction of combinations for the antimicrobial activity against the pig gut flora

In vitro simulations of the gastric and small intestinal fermentation with varying doses of carvacrol, thymol, eugenol and trans-cinnamaldehyde and binary combinations thereof were carried out to calculate dose–response equations for the antimicrobial effect against the main pig gut flora components. Interaction effects were evaluated following the isobole method. In gastric simulations eugenol and trans-cinnamaldehyde showed clearly less inhibitory activity towards lactobacilli than carvacrol and thymol, which could also be observed in the small intestinal simulations. The minimum concentration for carvacrol, thymol, eugenol and trans-cinnamaldehyde in small intestinal simulations to reduce the number of total anaerobic bacteria compared to control with a probability of 99.7% was 255, 258, 223 and 56 mg/l respectively. This strong activity of trans-cinnamaldehyde was due to its effect against coliform bacteria; a dose of 104 mg/l gave a reduction of 1 log CFU/ml vs. 371, 400 and 565 mg/l for carvacrol, thymol and eugenol respectively. Few combinations demonstrated synergism; most mixtures showed zero interaction or antagonism. Only carvacrol + thymol was slightly synergistic.     

Protection of laying hens against Salmonella Enteritidis by immunization with type 1 fimbriae

Eighteen chickens were immunized subcutaneously with purified type 1 fimbriae from Salmonella enterica serotype Enteritidis at 18 and 21 weeks of age. Evidence of IgG and IgA responses was found in the eggs and in the sera of the immunized hens. Three weeks later, immunized and non-immunized chickens (n=18) were challenged intravenously with 2x10(7) live Salmonella enterica serotype Enteritidis. There was no significant difference in the numbers of eggs laid by immunized and non-immunized birds. The percentage of Salmonella contaminated eggs was significantly higher in the non-immunized group than in the immunized group due to a higher percentage of contamination of the externally disinfected egg shells. There were no statistical differences in the percentages of contaminated yolks and egg whites between control and immunized birds. No differences in the number of colonizing bacteria could be found in the spleen nor in the liver between the immunized and the control groups throughout the experiment. Salmonella was cleared from the ovary of the immunized birds in the second week p.i., in contrast to the control birds where Salmonella was isolated till the third week after infection. Oviducts were significantly more infected in the control group than in the immunized group. Salmonella was cleared from the oviducts at 3 weeks p.i. in the immunized hens but not in the control hens. In conclusion, we demonstrated that the immunization of laying hens with type 1 fimbriae reduced the number of contaminated eggs and reduced the colonization of the reproductive organs.     

A Droplet Digital PCR Method for Detection of Newcastle Disease Virus

This experiment was conducted to establish a droplet digital PCR (ddPCR) method for the detection of Newcastle disease virus (NDV). A ddPCR method was developed, which the primers and probes were designed based on the conservative regions of F gene of NDV. The concentration of primer and probe, the annealing temperature in ddPCR reaction were optimized. The sensitivity, specificity and reproducibility of ddPCR method were evaluated. In results, the optimal primer concentration and the probe concentration were 900 and 250 nmol·L^-1, the optimum annealing temperature was 55℃. The detection limit of ddPCR method was 1.8 copies·μL^-1 with a good linear response, it had no cross reaction with other six viruses (include IBV), the coefficient of variation of sample repetition was 2.4%. All the results showed that NDV ddPCR was sensitive and specific, it was suitable for quantitative detection of clinical samples infected with NDV.     

Experiences and difficulties encountered during a course on veterinary public health with students of different nationalities

Veterinary public health (VPH) issues have received increased attention over the last few years as a result of the rising threat of emerging zoonoses (i.e., those due to globalized trade in animal and animal products and to changes in livestock production systems and the environment). The international dimension of VPH is gradually becoming recognized, and there is a growing need for veterinarians with experience in this field. In order to familiarize (future) veterinarians with the international dimension of VPH, the Department of Public Health and Food Safety of the Faculty of Veterinary Medicine, Utrecht University, has been organizing a course in Veterinary Public Health and Animal Production for over the last 10 years. This course has been intended for Dutch as well as foreign final-year veterinary students and recent veterinary graduates. By bringing together participants from different countries, the course reinforces the international dimension of the issues addressed through the exchange of experiences by the participants themselves. The present article provides information about this course on Veterinary Public Health (VPH): it discusses logistics, didactical approaches, the course program, and the use of information and communication technology (ICT). Special attention is given to the intercultural aspects of higher education, all of which play an important role in the efficient exchange of knowledge between lecturers and students. International courses are an important tool to enable participants to interact in a multicultural environment and address issues that demand international cooperation and a global public health focus.     

Urinary Biomarkers for Acute Kidney Injury in Dogs

Routinely, kidney dysfunction and decreased glomerular filtration rate (GFR) are diagnosed by the evaluation of changes in the serum creatinine (SCr) and blood urea nitrogen (BUN) concentrations. However, neither of these tests is sensitive or specific enough for the early diagnosis of impaired kidney function because they are both affected by other renal and nonrenal factors. Furthermore, kidney injury can be present in the absence of kidney dysfunction. Renal reserve enables normal GFR even when nephrons are damaged. Renal biomarkers, especially those present in urine, may be useful for the study of both acute and chronic nephropathies. The aim of this review is to describe the current status of urinary biomarkers as diagnostic tools for kidney injury in dogs with particular focus on acute kidney injury (AKI). The International Renal Interest Society (IRIS) canine AKI grading system and the implementation of urinary biomarkers in this system also are discussed. The discovery of novel urinary biomarkers has emerged from hypotheses about the pathophysiology of kidney injury, but few proteomic urine screening approaches have been described in dogs. Lack of standardization of biomarker assays further complicates the comparison of novel canine urinary biomarker validation results among studies. Future research should focus on novel biomarkers of renal origin and evaluate promising biomarkers in different clinical conditions. Validation of selected urinary biomarkers in the diagnosis of canine kidney diseases must include dogs with both renal and nonrenal diseases to evaluate their sensitivity, specificity as well as their negative and positive predictive values.     

Bovine blood neutrophil acyloxyacyl hydrolase (AOAH) activity during endotoxin and coliform mastitis

The dynamics of blood neutrophil acyloxyacyl hydrolase (AOAH) activity, the appearance of endotoxin (lipopolysaccharide, LPS) in blood and the role of blood neutrophil AOAH in the severity of Escherichia coli and endotoxin mastitis were investigated in early postpartum dairy cows experimentally challenged with either endotoxin (n = 6) or E. coli (n = 6). The AOAH activity of blood neutrophils started to decrease significantly at post challenge hours (PCH) 6-24 and 12-24 in the endotoxin and E. coli-challenged groups, respectively; it returned to pre-challenged values at PCH 48 in both endotoxin- and E. coli-challenged groups. The cows were classified as moderate and severe responders according to milk production loss in the non-challenged quarters at PCH 48. There were no severe responders in the endotoxin-challenged group. In the E. coli-challenged group, only 1 severe responder was identified. The pre-challenge neutrophil AOAH activity of the severe responder was approximately 30% lower than that of moderate responders. No LPS was detected in the plasma of endotoxin-challenged cows; neither was it found in the plasma of moderate responders in the E. coli-challenged group at any PCH. However, at PCH 6, a remarkable amount of LPS was detected in the plasma of the severe responder from the E. coli-challenged group. Furthermore, neutrophil AOAH activity was increased by approximately 70% in the severe responder at PCH 6, but it increased by only approximately 15% in moderate responders. This was followed by a decreased neutrophil AOAH activity at PCH 12-24 and 24-72 in moderate and severe responders, respectively; the decreased AOAH activity at those PCH was more pronounced in the severe responder. The pronounced decreased neutrophil AOAH activity during mastitis often coincided with extreme leukopenia, neutropenia and a maximal number of immature neutrophils in the blood. Our results demonstrate that a decrease in neutrophil AOAH activity results in the appearance of LPS in the blood, and low blood neutrophil deacylation activity could be considered as a risk factor for severe clinical coliform mastitis.