Field evaluation of the single intradermal cervical tuberculin test and the interferon-gamma assay for detection and eradication of bovine tuberculosis in Spain

A field comparison of the interferon-gamma (IFN-gamma) assay and the single intradermal cervical tuberculin (SICT) test for the diagnosis of bovine tuberculosis was conducted. A total of 1136 cattle belonging to 85 herds placed in 'Castilla y León' (northwestern Spain) were chosen, and 21 of these herds were subjected to the diagnostic assays two or three times at intervals of at least 4 months. All the animals positive to any of the tests were slaughtered and tuberculosis was confirmed by culture isolation method (CIM) and further identification by means of PCR. Only 10.6% of cattle reacted with the bovine PPD in the SICT test, a percentage that increased to 12.8% in the IFN-gamma assay. The sensitivity of the IFN-gamma assay compared to CIM was shown to be higher (84.9%) than that of the SICT test (80.2%), but the combination of both tests offered the highest sensitivity (92.9%). The number of false positive reactors (those animals in which CIM was negative) was considerably higher for the IFN-gamma assay than for the SICT test and, conversely, the number of false negative animals (M. bovis isolation but negative immunological result) was higher for the skin test than for the interferon assay. In the herds tested twice, tuberculosis was eradicated after the second cycle of testing in 50%, and in 75% after the third cycle in herds tested three times. The combination of these two techniques instead of separately seems, therefore, to be useful in eradication programmes against bovine tuberculosis.     

Determination and persistence of imazalil in post-harvest-treated apples during cold storage

A method to determine imazalil (allyl 1 -(2, 4-dichlorophenyl)-2-imidazol- 1-ylethyl ether) on apples using high performance liquid chromatography is described. After harvest, fruits were immersed in an aqueous suspension (1 g litre-1) and cold stored (0-2°C) at 85-90% r.h. Samples were taken monthly and imazalil was determined in the peel, outer pulp, inner pulp and on whole fruit. Residues were 3.8-4.9 mg imazalil kg-1 and decreased during storage. Most imazalil was found on the peel and amounts in the pulp decreased toward the core.     

Structure-activity relationship study of alkynyl ether insecticide synergists and the development of MB-599 (verbutin)

Structure-activity relationships of aryl alkynyl synergists of the general formula of Ar-Q-R, where Q represents a bridging structure, were studied using a standardised testing system and Relative Potency values. Ethers, esters, oxime ethers, amides and amines were prepared and evaluated. The length of the R-alkynyl chain, the role of the bridge and the substitution of the aromatic ring were examined systematically. The most potent compounds possessed an aromatic ring connected via a bridge of three atoms to an alkynyl chain, forming together a linear side-chain of six atoms. Several highly potent compounds were synthesised of which one (MB-599; proposed common name verbutin) was selected for development as a selective insecticide synergist in crop protection. Its high potential at practical insecticide:synergist ratios makes possible the reduction of the total amount of insect-control chemicals applied, and its use as an additive to produce new formulations of existing insecticides makes it highly advantageous in resistance management, giving a new tool to sustain the effectiveness of a wide range of insecticides. A product containing a (1+1) mixture of verbutin and beta-cypermethrin was launched in Hungary in 2002.     

Evaluation of the local analgesic effect of ketamine in the palmar digital nerve block at the base of the proximal sesamoid (abaxial sesamoid block) in horses

OBJECTIVE: To evaluate the local analgesic effect of ketamine in a palmar digital nerve block at the base of the proximal sesamoid (abaxial sesamoid block) in horses. ANIMALS: 36 mature healthy Andalusian horses. PROCEDURE: Horses were randomly assigned to 4 groups of 9 horses each and received an abaxial sesamoid block in a randomly chosen forelimb with 1 of the following: saline (0.9% NaCl) solution, 1% ketamine solution, 2% ketamine solution, or 3% ketamine solution. To determine analgesia, the radiant heat lamp-hoof withdrawal model was used as a noxious thermal stimulus. Before each nerve block, baseline hoof withdrawal reflex latency (HWRL, time between lamp illumination and withdrawal of the hoof) was determined; after the nerve block, local analgesic effects were determined by measuring HWRL at 2 and 5 minutes after injection and then every 5 minutes for a total period of 1 hour. RESULTS: Significant differences in HWRL were found between baseline values and values at 2 to 15 minutes following a nerve block with ketamine. Significant differences were found between HWRL values at every time point from 2 to 10 minutes following a nerve block with saline solution, compared with 1 or 2% ketamine solution. Similarly, significant differences were found between HWRL values at every time point from 2 to 15 minutes following a nerve block with saline solution, compared with 3% ketamine solution. CONCLUSIONS AND CLINICAL RELEVANCE: Abaxial sesamoid block with ketamine ensures adequate analgesia in horses with an onset of action of 2 minutes and a maximal duration of action of 15 minutes.     

Palaeoparasitological analysis of guinea pig mummies of the Chiribaya culture,Moquegua Valley,Peru

During several excavations of archeological sites in the Moquegua Valley, Southern Peru, various guinea pig mummies (Cavia aperea f. porcellus) have been discovered. They belong to the Chiribaya Culture (900-1100 AD). The outstanding preservation of the mummified animals gave rise to the idea of a paleoparasitological analysis. In the fur, numerous well preserved ectoparasites (lice, fleas, mites) could be recovered. Generally, ectoparasite remains are rarely found among archeological material. This is the first account of an extensive ectoparasitological analysis of animal mummies in Peru. A modified technique for recovery and preservation of the ectoparasites has been developed.     

Co-feeding studies of ticks infected with Anaplasma marginale

Ticks often cluster at preferred feeding sites on hosts, and the co-feeding of ticks at the same site has been shown to increase feeding success and the transmission of some pathogens. While the major route of infection of ticks with pathogens is via the bloodmeal during feeding on a parasitemic host, non-systemic transmission of viruses and spirochetes has been shown to occur from infected to uninfected ticks at common feeding sites on uninfected hosts. In this research, two separate studies were done using the tick-borne rickettsial pathogen of cattle, Anaplasma marginale. In one study we tested whether A. marginale could be transmitted non-systemically from infected to uninfected Dermacentor variabilis males while co-feeding on rabbits. Infection of ticks was determined by allowing them to transmission feed on susceptible cattle and by DNA probe and microscopy studies on salivary glands. In the second study, we tested whether the co-feeding of male and female ticks on parasitemic cattle would increase the acquisition and development of A. marginale in males. A. marginale infections in salivary glands were determined by quantitative PCR after the ticks were allowed to transmission feed on susceptible cattle. Non-systemic transmission of A. marginale did not occur from infected and uninfected ticks that fed at the same site on rabbits and, therefore, does not appear to be a means of A. marginale transmission. A. marginale infections in male ticks were not increased while co-feeding with females. Thus, co-feeding of adult Dermacentor spp. does not appear to influence the dynamics of A. marginale transmission.     

Plasma and synovial fluid endothelin-1 and nitric oxide concentrations in horses with and without joint disease

OBJECTIVE: To compare plasma and synovial fluid endothelin-1 (ET-1) and nitric oxide (NO) concentrations in clinically normal horses and horses with joint disease. ANIMALS: 36 horses with joint disease, and 15 horses without joint disease. PROCEDURE: Horses with joint disease were assigned to 1 of the 3 groups (ie, synovitis, degenerative joint disease [DJD], or joint sepsis groups) on the basis of findings on clinical and radiographic examination and synovial fluid analysis. Endothelin-1 and NO concentrations were measured in plasma from blood samples, collected from the jugular vein and ipsilateral cephalic or saphenous vein of the limb with an affected or unaffected joint, as well as in synovial fluid samples obtained via arthrocentesis from the involved joint. RESULTS: Plasma ET-1 concentrations between affected and unaffected groups were not significantly different. Median concentration and concentration range of ET-1 in synovial fluid obtained from the joint sepsis group (35.830 pg/mL, 7926 to 86.614 pg/mL; n = 7) were significantly greater than values from the synovitis (17.531 pg/mL, 0.01 to 46.908 pg/mL; 18), DJD (22.858 pg/mL, 0.01 to 49.990 pg/mL; 10), and unaffected (10.547 pg/mL, 0.01 to 35.927 pg/mL; 10) groups. Plasma and synovial fluid NO concentrations between affected and unaffected groups were not significantly different. CONCLUSIONS AND CLINICAL RELEVANCE: Endothelin-1 is locally synthesized in the joints of horses with various types of joint disease. Synovial fluid concentrations of ET-1 varied among horses with joint disease, with concentrations significantly higher in the synovial fluid of horses with joint sepsis. These results indicate that ET-1 may play a role in the pathophysiologic mechanism of joint disease in horses.     

Development of Spinitectus inermis (Nematoda: Cystidicolidae), a parasite of eel,Anguilla anguilla,in Europe

The development of the nematode Spinitectus inermis (Zeder, 1800), a parasite of the stomach of eels, Anguilla anguilla (L.) in Europe, was experimentally studied. Mayfly nymphs Caenis macrura, Ecdyonurus dispar, Heptagenia sulphurea, Potamanthus luteus and Seratella ignita from Portugal and the Czech Republic were found to serve as experimental intermediate hosts. After ingestion of the nematode eggs by the mayfly nymphs, the toothed first-stage larvae were released and penetrated into the body cavity of the intermediate host. There they moulted twice (on day 4 and 6 post infection [p.i.] at water temperatures of 20-25 degrees C), attaining the third infective stage. The definitive host, A. anguilla, undoubtedly acquires infection by feeding on mayfly nymphs harbouring infective-stage larvae. In an experimentally infected eel, the fourth-stage larva undergoing the third moult was observed 28 days p.i. at water temperature of 20 degrees C. The larval stages, including moulting forms, are described and illustrated. The prepatent period of S. inermis is estimated to be about two months.