Trends in antimicrobial susceptibility in relation to antimicrobial usage and presence of resistance genes in Staphylococcus hyicus isolated from exudative epidermitis in pigs

From 1996 to 2001 a total of 467 Staphylococcus hyicus isolates from exudative epidermitis (EE) in pigs in Denmark were examined for susceptibility to 13 different antimicrobial agents. The presence of selected genes encoding macrolide (erm(A), erm(B) and erm(C)), penicillin (blaZ), streptogramin (vat, vga, vga(B), vat(B), vat(D) and vat(E)), streptomycin (aadE) and tetracycline resistance (tet(K), tet(L), tet(M) and tet(O)) were determined in selected isolates.The occurrence of erythromycin resistance increased from 33% in 1996 to a maximum of 62% in 1997 and decreased to 26% in 2001. Resistance to sulphametazole increased from 17% in 1996 to 30% in 1998 but has since decreased to 4% in 2001. Resistance to trimethoprim increased to 51% in 1997 and decreased to 21% in 2001. Resistance to tetracycline (21-31%) remained relatively constant during 1996-2000, but increased to 47% in 2001. Resistance to penicillin (54-75%) streptomycin (33-53%) and tetracycline (21-47%) remained relatively constant over the time investigated.All 48 penicillin resistant isolates examined contained the blaZ gene and 40 (85%) of the streptomycin resistant isolates the aadE gene. It was not possible to detect any streptogramin resistance gene in four streptogramin resistant isolates. Of the 55 erythromycin resistant isolates examined, five contained erm(A), 13 erm(B), 35 erm(C) and two both erm(A) and erm(C). The presence of erm(B) was confirmed by hybridization to plasmid profiles in all 13 PCR-positive isolates. Of 52 tetracycline resistant isolates examined, two contained tet(L), 38 tet(K) and 12 both tet(K) and tet(L).     

Transmission of Actinobacillus pleuropneumoniae in pigs under field-like conditions: emphasis on tonsillar colonisation and passively acquired colostral antibodies

The objectives of this study were to elucidate at which age tonsillar colonisation by Actinobacillus pleuropneumoniae occurs in pigs and relate this occurrence to the presence of colostral antibodies to A. pleuropneumoniae. The infection patterns were studied in an isolated cohort of pigs, which consisted of the offspring from five sows originating from a conventional pig herd. The sows were transferred to isolated research facilities before farrowing. A. pleuropneumoniae was detected on the tonsils of all sows. After a nursing period of 3 weeks, the pigs were weaned and reared isolated from other pigs until slaughter. The pigs were examined repeatedly for the presence of A. pleuropneumoniae on the tonsils and for antibodies to A. pleuropneumoniae using bacteriological and serological techniques, respectively.A. pleuropneumoniae was detected in the tonsils of one pig as early as 11 days after birth, showing that A. pleuropneumoniae can be transmitted from sow to offspring during a 3-week nursing period. The cumulative proportion of pigs carrying A. pleuropneumoniae in their tonsils increased significantly between the age of 4-12 weeks. This age period corresponded to the age at which the proportion of pigs with detectable levels of colostral antibodies to the different serotypes of A. pleuropneumoniae was declining. Since these two events take place in the same age period, we expect a possible biological association between the level of the passive immunity and the degree of tonsillar colonisation. The median duration of tonsillar colonisation was estimated to approximately 7-8 weeks.     

Risk factors for metritis in Danish dairy cows

A retrospective longitudinal study of metritis was conducted in Denmark on data collected during 1993-1994. Data on herd size, breed, parity, and treatment of disease were obtained from the Danish Cattle Database. Management and production-facility data were collected using a questionnaire, conducted as a telephone interview in 1994. The study included 2144 herds from three regions in Denmark (102,060 cows). Herd-level variables included were: herd size, housing, flooring, grazing, calving measures, and calving supervision. Cow-level variables were: parity, breed, calving season and whether the cow had been treated by a veterinarian for dystocia or the diseases: retained placenta, reproductive disease, ketosis, milk fever, or dry cow mastitis.Marginal multivariable logistic-regression analyses were performed. The cow with highest odds of metritis was a first or greater than or equal to third parity cow, of large breed, that calved during November-April, in a zero-grazing herd. The cow had been treated for dystocia, retained placenta, and at least one other reproductive disease, but not for ketosis.     

A meta-analysis comparing the effect of vaccines against Mycoplasma hyopneumoniae on daily weight gain in pigs

Our aims were to evaluate the published literature concerning the effect of swine vaccination against Mycoplasma hyopneumoniae on the average daily weight gain (ADWG). This was done by re-evaluating the influence of selected factors on ADWG by a meta-analysis of published studies from 1991 to 1999, fulfilling certain inclusion criteria.With ADWG as the outcome, an analysis of variance was performed for such variables as treatment, vaccination schedule, age during study, housing system and publication quality. Each clinical trial was considered as a random effect and the numbers of pigs in each trial were weightings.Of 63 published studies, 16 describing three commercial vaccines fulfilled the criteria for the meta-analysis. Due to few studies with one of the vaccines (n=3), only two vaccines were included.Vaccinated pigs gained an average of 592g (S.E.=15) with Stellamune and 590g (S.E.=15) with Suvaxyne compared to non-vaccinated pigs that gained an average of 569g (S.E.=14)(P<0.01) when adjusted for age during the study. Vaccine type, vaccination schedule, housing system and publication quality were not significantly associated with ADWG.     

Maximum-likelihood estimation of sensitivity and specificity of ELISAs and faecal culture for diagnosis of paratuberculosis

The accuracy of three diagnostic tests for paratuberculosis was evaluated using maximum-likelihood estimation of sensitivity and specificity. We also explored the variety of estimates that can be obtained if the tests are to be used in populations of different composition with regard to infection and disease states. Two enzyme-linked immunosorbent assays (ELISAs) were evaluated separately with the faecal culture (FC). The study was carried out as a cross-sectional field study to cover all likely states of infection with Mycobacterium avium subsp. paratuberculosis.The three basic assumptions for the maximum-likelihood technique were evaluated to validate the results. Our accuracy estimates for the ELISAs were not very different from those previously published, but those for faecal culture differed if a different cut-off value was chosen for the ELISA. If faecal culture was used for screening in a Danish dairy region where the median ELISA reading was a measure of the general disease situation, the sensitivity of the faecal culture was 20-25%. If faecal culture was used as a confirmatory test on cows with a high ELISA reading (and thus high level of antibodies), the sensitivity of the faecal culture would be in the range 60-70%. These results emphasise the importance of the composition of a target population before selecting a specific diagnostic test for a given purpose. We concluded that faecal culture is useful for confirmation but not for screening purposes.     

Estimation of sensitivity and specificity of diagnostic tests and disease prevalence when the true disease state is unknown

The performance of a new diagnostic test is frequently evaluated by comparison to a perfect reference test (i.e. a gold standard). In many instances, however, a reference test is less than perfect. In this paper, we review methods for estimation of the accuracy of a diagnostic test when an imperfect reference test with known classification errors is available. Furthermore, we focus our presentation on available methods of estimation of test characteristics when the sensitivity and specificity of both tests are unknown. We present some of the available statistical methods for estimation of the accuracy of diagnostic tests when a reference test does not exist (including maximum likelihood estimation and Bayesian inference). We illustrate the application of the described methods using data from an evaluation of a nested polymerase chain reaction and microscopic examination of kidney imprints for detection of Nucleospora salmonis in rainbow trout.     

Screening for infection of Trichinella in red fox (Vulpes vulpes) in Denmark

A total of 6141 foxes (Vulpes vulpes) were examined for infection with Trichinella. The foxes were killed in Denmark during the hunting season 1995-1996 and 1997-1998; 3133 and 3008, respectively. Foxes included in the investigation came from throughout the country with the exception of the island of Bornholm. The right foreleg from each fox was submitted for investigation. The legs were stored at -20 degrees C for 3-10 months prior to examination. Following thawing, muscle tissue (10 g) from each leg was examined by trichinoscopy and by a pepsin-HCl digestion technique. In 1995-1996, three foxes were found positive corresponding to a prevalence of 0.001. Each of the infected foxes harboured an extremely low infection, i.e. about one larva per 10 g muscle tissue. It was not possible to obtain sufficient larval material for species identification. All three foxes were shot in the vicinity of a small village in the north-western part of Denmark. In 1997-1998 no Trichinella cases were found. The results, compared with previous studies, indicate that the prevalence of infection of Trichinella sp. among wild living foxes in Denmark is very low. This is further supported by the fact, that no infection of Trichinella sp. has been found in slaughtered pigs in Denmark for more than 65 years, which suggests that the infection pressure is very low. Considering the facts above we conclude that the risk of Trichinella infections is negligible in intensive indoor pig production units in Denmark whereas high local prevalence of Trichinella infections in the wildlife might constitute a serious risk for the expanding outdoor pig production.     

Use of a migration assay for the separation of adult pyrantel-susceptible and -resistant oesophagostomum dentatum

A migration assay was used to separate a pyrantel-susceptible and -resistant isolate of the pig nematode, Oesophagostomum dentatum. The experiment had three steps. In the first step, LD(50) values for pyrantel of the two isolates in the assay were established. In the second step, susceptible and resistant worms were mixed in various proportions prior to exposure to a fixed concentration of pyrantel and thereafter assayed. The inhibition of migration showed to be linearly correlated with the proportion of resistant worms in a sample. In step three, this line was used as a standard curve to predict the number of resistant worms in samples from pigs infected with mixed samples of susceptible and resistant larvae.     

Antibody dynamics in BRSV-infected Danish dairy herds as determined by isotype-specific immunoglobulins

Using specific ELISAs, antibody levels of four different isotypes to bovine respiratory syncytial virus (BRSV) were determined in calves, following experimental BRSV infection.Most calves experienced an increase in the specific IgM and IgG1 titres about 6-10 days after infection with BRSV. The IgM titre was transient showing positive titres for only 5-10 days, while specific IgG1 was present for a longer time. IgA was detected concomitantly with IgM but at a lower level. Production of IgG2 anti-BRSV antibodies was detected from 3 weeks after infection.In two closed herds, repeated blood samplings were performed on young stock to analyse maternal immunity. The passively transferred antibodies were mainly of the IgG1 isotype and the half-life of IgG1 to BRSV was estimated to be 26.6 days. One of the herds had an outbreak of enzootic pneumonia, diagnosed to be caused by BRSV. Furthermore, another herd with acute BRSV was followed by weekly blood samples in six calves; in both herds IgM and IgG1 was detected shortly after the appearance of clinical signs. Serum samples from 50 Danish dairy herds (453 samples) were tested for immunoglobulins of the isotypes IgG1, IgG2 and IgM. The presence of antibodies to BRSV was widespread and more than 54% of the samples had BRSV antibodies of both the IgG1 and IgG2 isotypes indicating a high herd prevalence to BRSV. Test samples from two herds out of 50 were free from all isotypes to BRSV.     

Sap flow in papaya plants: Laboratory calibrations and relationships with gas exchanges under field conditions

In papaya plants a study to quantify the water flow through the trunk is important for to promote a good water management in commercial orchard. The objective was to study the relationship between water flow through the trunk and temperature measurements determined by probes inserted in the papaya plant stem in laboratory. In addition, was possible to study the relationship between sap flow and instantaneous gas exchange in field conditions. We constructed an instrument that maintained a stable water flux through a 0.30 m stem section with a constant pressure, simulating the xylem sap flow through the stem. Water flux was adjusted by varying pressure of water in the stem section. The mathematical model used to fit the relation between K (Granier heat coefficient) values and sap flow density was the exponential model: u = 0.5511 × K^1.9104. Field studies was conducted in a commercial orchard located in North of the State of Rio de Janeiro, with 12 plants in October 2002, and eight plants in January 2003. We verified that instantaneous transpiration, measured by a portable system of gas exchange (porometry), presented a good (R² = 0.75) positive relationship with xylem sap flow. Estimates of papaya sap flow can be obtained by scaling portable photosynthesis system measurements with exposed leaves, however the relationship is non-linear in higher instantaneous transpiration rates. The causes of the non-linear relationship in higher transpiration are discussed. In addition, was possible to obtain a good (R² = 0.76) relationship between net photosynthesis rate and xylem sap flow in papaya field-grown.