Detection of residues of the coccidiostat diclazuril in poultry tissues by liquid chromatography-tandem mass spectrometry after withdrawal of medicated feed

A liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for the quantitative determination of diclazuril in poultry tissues and feed is presented. A simple clean up with an organic solvent was carried out. A reversed-phase C(18) column was used for the high-performance liquid chromatography (HPLC) to separate the analyte with a gradient of acetonitrile and water as mobile phase. The precursor ions produced by electrospray negative ionization were selected for collisional dissociation. Validation of the methods was performed based on Commission Decision 2002/657/EC (Off. J. Eur. Communities 2002, L221, 8-36). For the detection of diclazuril in poultry meat, the decision limit was found to be 0.5 microg/kg. An animal experiment was set up in which 70 chickens were held for 6 weeks. From day 22 until day 32, they were fed feed containing 730 microg/kg diclazuril. From day 33 until day 42, every day six chickens were slaughtered, and breast, thigh, and liver were analyzed. Average steady-state concentrations of 94, 135, and 722 microg/kg in breast, thigh, and liver were obtained, respectively. Nine days after withdrawal of the medicated feed, diclazuril was still present in the different sample types.     

Infectious bursal disease virus (IBDV) induces apoptosis in chicken B cells

The ability of infectious bursal disease virus (IBDV) serotypes 1 and 2, and the role of VP4 of both serotypes as well as the capacity of three IBDV intermediate serotype 1-specific vaccine strains to induce apoptosis in a chicken B-lymphocyte cell line, DT40, were investigated using the TUNEL technique. It was observed that IBDV serotype 1 infected the DT40 cell line and directly induced apoptosis. In contrast, the non-pathogenic serotype 2 neither infected nor induced apoptosis, but was able to reduce the serotype 1-induced apoptosis when the two viruses were present in combination. VP4 of both serotypes did not induce apoptosis. IBDV VP2 of serotype 2 induced apoptosis in the same proportion and intensity as VP2 of serotype 1. IBDV intermediate vaccines varied in their ability to induce apoptosis in the DT40 cell line, which was also decreased-delayed in presence of serotype 2 IBDV. We hypothesize that both serotypes compete for the same receptor in DT-40 cells, and suggest that IBDV-induced apoptosis is a multistep process involving virus replication, protein expression, and release of virions.     

Diagnosis of cystic echinococcosis on sheep farms in the south of Argentina: areas with a control program

In 2000 Guarnera et al. proposed using ELISA in canine faeces collected from the ground to detect dogs infected with Echinococcus granulosus, thus determining sheep farms with active transmission. The objective was to evaluate the prevalence of E. granulosus infection in sheep farms of the Patagonia. Sheep farms were randomly selected in the Provinces of Río Negro, Chubut, Neuquén, Santa Cruz and Tierra del Fuego (areas with control programs) and La Pampa (comparison area). From one to three samples of fecal matter were obtained for each sheep farm, which were processed by means of copro-ELISA test with confirmation of positive samples by copro-Western blot. A total of 1042 samples were obtained from 352 sheep farms, 26 (7.3%) proving positive. Of these 5 (6.3%) were from La Pampa, 9 (13.8%) from Neuquén, 4 (4.7%) from Río Negro, 2 (2.9%) from Chubut, 1 (5.9%) from Santa Cruz and 5 (13.9%) from Tierra del Fuego. The identification of parasitized dogs is an essential activity upon which rests the strategy of control and surveillance. Arecoline tests or coproantigen test with fecal matter obtained directly from the dog contribute information on individual prevalence, while the use of coproantigens detected in ground-collected samples transfers the dog unit of observation to units of greater epidemiological value. In the present experience, the technique employed seems promising for its application in systems of epidemiological surveillance of cystic echinococcosis and in drawing a baseline on which to measure the progress of control programs in the Argentine Patagonia in subsequent years.     

Multiple-criteria decision-making techniques and their role in livestock ration formulation

The livestock ration formulation problem is postulated within the framework of multiple-criteria decision-making techniques. This exercise is motivated by the fact that the ordinary least-cost approach can, and does, generate solutions that either cannot be implemented or supply nutritionally undesirable levels of various nutrients. This originates from using cost as the criterion for selecting the ingredients of the diet. But in fact, diet formulation problems involve several criteria and should, therefore, be solved using techniques designed for modelling such problems. This paper is an attempt at introducing these techniques to agricultural systems modellers and then demonstrating their use in livestock ration formulation. The multiple-criteria decision-making techniques covered include goal programming and its variants such as weighted and lexicographic approaches and multiple-objective programming.     

Failure to establish infection with Tetratrichomonas sp. in the reproductive tracts of heifers and bulls

Experimental infection of the reproductive tracts of heifers and bulls with Tetratrichomonas sp. isolated from preputial smegma of virgin bulls was attempted. Nine heifers and four bulls were challenged by inoculation of 7 x 10(6) Tetratrichomonas sp. into the vaginal lumen and preputial cavity, respectively. Vaginal mucus and preputial smegma samples were collected and cultured for Tetratrichomonas sp. Heifers were slaughtered in groups of three at 2, 9 and 21 days after inoculation. Two heifers and two bulls infected with Tritrichomonas foetus and two uninfected heifers were used as controls for the model infection. Tetratrichomonas sp. were only isolated in vaginal mucus of 7/9 inoculated heifers at 6h post-inoculation, and genital secretions taken at slaughter time from vagina, uterus and oviduct were cultural negative. Bulls challenged with Tetratrichomonas sp. remained cultural negative. Since Tetratrichomonas sp. survived only a few hours in the female genitalia and did not survive in the male genitalia after experimental challenge, Tetratrichomonas sp. did not colonize the genital tract. These were likely trichomonads from the digestive tract. Collection of clean samples without fecal contamination from the reproductive tract is proposed as a measure to avoid Tetratrichomonas sp. transitory genital infection.     

Virulence-associated genes in Escherichia coli isolates from poultry with colibacillosis: correction

Several virulence genes of avian Escherichia coli were detected in 200 colibacillosis isolates from our region by PCR. However, the genes sfaDE and facA were not detected in that study. In this work we correct those data, showing by colony hybridization that sfaDE and facA are present in 40% and 30% of those isolates, respectively.     

Beta-glucan enhancement of T cell IFNgamma response in swine

Beta-glucan has been shown to enhance anti-tumor and anti-infection functions in animals. Pigs at 4 months of age were infected with porcine reproductive and respiratory syndrome virus (PRRSV), and peripheral blood monocytes (PBMC) were isolated for the detection of interferon gamma (IFNgamma)-producing cells. We found that soluble high molecular weight beta-glucan could increase IFNgamma-producing cell frequency in a dose-dependent manner in the enzyme-linked immunospot assay (ELISPOT) in the absence of antigenic restimulation. A concentration as low as 1.6 microg/ml gave a significant increase and a similarly high enhancement was achieved at concentrations from 3.2 to 100 microg/ml. In PRRSV-specific IFNgamma ELISPOT, soluble beta-glucan elicited increased PRRSV-specific responses at concentrations from 3.2 to 50 microg/ml, but not at 100 microg/ml, whereas insoluble beta-glucan had no effect. Soluble beta-glucan augmented the porcine cellular immune response in an antigen-independent fashion, whereas insoluble beta-glucan had no activity. This finding suggests that soluble beta-glucan may enhance innate antiviral immunity against PRRSV.     

Quantitative assessment of regional right ventricular myocardial velocities in awake dogs by Doppler tissue imaging: repeatability, reproducibility, effect of body weight and breed, and comparison with left ventricular myocardial velocities

Right ventricular myocardial (RVM) motion is poorly documented. The objective of this study was to determine the variability of RVM velocities by tissue Doppler imaging (TDI) in healthy dogs (study 1), to analyze RVM motion in a large healthy canine population (study 2), and to compare the results with those obtained for the left ventricular free wall. Six healthy Beagle Dogs were monitored in study 1, and 64 healthy dogs of 14 different breeds were monitored in study 2. Velocities were recorded in 2 segments (basal and apical) of the right and left myocardial walls. In study 1, 36 TDI examinations were performed for 4 days, whereas a single TDI examination was performed on each dog in study 2. All velocity profiles included 1 positive systolic wave and 2 negative diastolic waves. The lowest intraday and interday coefficient of variation values of the right TDI variables were observed at the base (3.5-16.1%). The variability of the right apical velocities was much higher, with most coefficient of variation values > 15%. RVM velocities were higher in the basal than in the apical segments (P < .001) and were higher than the left velocities of the corresponding segment (P < .01). Body weight and breed had an effect on only a few right and left TDI variables. TDI provides a repeatable and reproducible method for evaluating basal RV function in the dog. These data also demonstrate the heterogeneity of the myocardial velocities between the left and the right ventricles and between the base and the apex.     

Longitudinal left ventricular myocardial dysfunction assessed by 2D colour tissue Doppler imaging in a dog with systemic hypertension and severe arteriosclerosis

A 12-year-old sexually intact male Vendee Griffon Basset was presented for acute pulmonary oedema. Severe systemic systolic arterial hypertension (SAH) was diagnosed (290 mmHg). Despite blood and abdominal ultrasound tests, the underlying cause of the systemic hypertension could not be determined, and primary SAH was therefore suspected. Conventional echocardiography showed eccentric left ventricular hypertrophy with normal fractional shortening. Despite this apparent normal systolic function, 2D colour tissue Doppler imaging (TDI) identified a marked longitudinal systolic left ventricular myocardial alteration, whereas radial function was still preserved. Three months later, the dog underwent euthanasia because of an acute episode of distal aortic thromboembolism. Necropsy revealed severe aortic and iliac arteriosclerosis. SAH related to arteriosclerosis is a common finding in humans, but has not been previously described in dogs. Moreover, its consequence on longitudinal myocardial function using TDI has never been documented before in this species.     

Ultrastructural and immunocytochemical evaluation of the effects of extracorporeal shock wave treatment in the hind limbs of horses with experimentally induced suspensory ligament desmitis

OBJECTIVE: To evaluate the effects of extracorporeal shock wave therapy (ESWT) on affected ligaments in the hind limbs of horses with experimentally induced suspensory ligament desmitis by use of ultrasonographic, ultrastructural, and immunocytochemical techniques. ANIMALS: 10 horses. PROCEDURE: Suspensory ligament desmitis was induced in both hind limbs of each horse by use of 2 collagenase injections (administered 2 weeks apart) in each suspensory ligament. Two weeks after the second injection, the right hind limb of each horse was treated with ESWT (3 treatments at 3-week intervals); the left hind limb was not treated (control limb). Periodically during the study, the healing process was monitored ultrasonographically and the proportions of ligaments affected with lesions were assessed. Four weeks after the last ESWT treatment, biopsy specimens were collected from all ligaments for ultrastructural evaluation and immunocytochemical analysis of transforming growth factor beta-1. RESULTS: The difference in the proportion of the lesion-affected ligament between ESWT-treated and control limbs was significant (P < 0.05) from 3 weeks after the second ESWT treatment to the end of the study. Compared with control ligaments, ESWT-treated ligaments had more small, newly formed collagen fibrils and greater expression of transforming growth factor beta-1 4 weeks after the last ESWT treatment was administered. CONCLUSIONS AND CLINICAL RELEVANCE: Results have indicated that ESWT appears to facilitate the healing process in horses with experimentally induced hind limb suspensory ligament desmitis.