Sven Erik Lind Jorsal

Detection of Lawsonia intracellularis, Serpulina hyodysenteriae, weakly beta-haemolytic intestinal spirochaetes, Salmonella enterica, and haemolytic Escherichia coli from swine herds with and without diarrhoea among growing pigs.

A polymerase chain reaction (PCR) was optimized to detect Lawsonia intracellularis in faeces from naturally infected pigs. By combining a boiling procedure to extract DNA and a nested PCR procedure, a detection limit at 2 x 10(2) bacterial cells per gram of faeces was achieved. The optimized PCR was used together with conventional culture techniques to detect Serpulina hyodysenteriae, weakly beta-haemolytic intestinal spirochaetes (WBHIS), Salmonella enterica, and haemolytic Escherichia coli, in a case control study to examine selected risk factors for the development of diarrhoea in growing pigs. Herds with diarrhoea were selected as cases and randomly chosen herds without diarrhoea were chosen as controls. Infection with L. intracellularis significantly enhanced the chance of diarrhoea. S. hyodysenteriae, WBHIS group IV (Serpulina pilosicoli), and S. enterica were isolated only from case herds which indicate that these species may influence the development of diarrhoea. In addition, herd-type had a significant impact, that is specific pathogen-free herds showed an odds ratio at 0.2 relative to conventional herds for the development of diarrhoea.

Prevalence of intestinal pathogens in Danish finishing pig herds

Our aim was to determine the prevalence of the intestinal bacteria: Lawsonia intracellularis, Brachyspira hyodysenteriae, Serpulina intermedia, Brachyspira innocens, Brachyspira pilosicoli, pathogenic Escherichia coli (serogroups O138, O139, O141 and O149) and Salmonella enterica in Danish finishing pig herds. A total of 79 herds was randomly selected and visited during 1998. From each herd, 20 faecal samples were collected from individual pigs weighing 30-50kg. Furthermore, 10 pooled pen samples were collected and examined for S. enterica. In total, 1580 faecal samples and 790 pen samples were collected and examined by polymerase chain reaction (PCR) or culture. L. intracellularis was found in 74 herds (93.7%), B. hyodysenteriae in two herds (2.5%), S. intermedia in 10 herds (12. 7%), B. innocens in 27 herds (34.2%), B. pilosicoli in 15 herds (19. 0%), pathogenic E. coli in 19 herds (24.1%) and S. enterica in eight herds (10.1%). The within-herd prevalences of L. intracellularis and B. hyodysenteriae were 25-30%; the within-herd prevalences of the other agents were 5-10%. Three herds (4%) were not infected with any of the bacteria and 25 herds (32%) were only infected with L. intracellularis.

Prevalence and diversity of Arcobacter spp. isolated from the internal organs of spontaneous porcine abortions in Denmark

A study was conducted to determine the prevalence and possible significance of campylobacteria in pig abortions in Denmark. Surface-cauterised liver and kidney samples from 55 aborted pig fetuses submitted to the Danish Veterinary Laboratory were taken and a sensitive isolation procedure used to examine pooled tissue samples for Campylobacter, Arcobacter and Helicobacter spp. Routine microbiological, immunological, and histopathological examinations were also performed to identify concurrent infections or histopathological changes. The abortions tested negative for established abortifacient pathogens (Brucella, Leptospira, PPV, PRRSV), but Arcobacter spp. were recovered from 23/55 abortions. Co-infections with Streptococcus suis, Escherichia coli, and haemolytic streptococci were observed in 7/23 Arcobacter-positive fetuses, and in 4/32 Arcobacter-negative fetuses. Histopathological analyses identified placentitis, pneumonia, hepatitis and encephalitis among the study group. However, no obvious pathologic features were solely associated with Arcobacter-positive cases, nor were Arcobacter-like bacteria observed in tissue samples. Protein profile analyses of the 27 Arcobacter isolates identified 11 as A. cryaerophilus and 10 as A. skirrowii. Six strains could not be classified into any existing species and were phenotypically distinct, thus, potentially representing at least one new species. The identification results showed that multiple taxa could be found in a single fetus, and in distinct aborted fetuses from a single sow. The high prevalence of arcobacters in Danish pig abortions may account for at least some of the >90% of cases in which no established abortifacient agent is detected, but further studies are needed to define the role of each species, especially where co-infections with other bacteria are present.

Serological characterization of Actinobacillus pleuropneumoniae biotype 2 strains isolated from pigs in two Danish herds.

Eight Actinobacillus pleuropneumoniae biotype 2 strains were isolated in pure culture from lungs of pigs originating from two Danish herds with growing and finishing pigs. The antigenic properties were studied by indirect haemagglutination (IHA) and immunodiffusion (ID) tests using soluble surface antigens and by enzyme-linked immunosorbent assays (ELISA) using capsular enriched fractions and LPS. In all tests the strains proved antigenically homogeneous and serologically distinct from the known biotype 1 and 2 serotypes. Thus, the strains represent a new serotype which is provisionally proposed as biotype 2 serotype 14.

Serological characterization and antimicrobial susceptibility of Streptococcus suis isolates from diagnostic samples in Denmark during 1995 and 1996.

At the Danish Veterinary Laboratory Streptococcus suis infections in pigs were diagnosed in 114 cases in 1995 and in 151 cases in 1996. Isolates were serotyped using specific antisera against type 1 through 28 and a total of 67 cases from 1995 and 113 cases in 1996 were tested for resistance to 11 antimicrobial agents. The majority of cases were lung diseases (57%), followed by septicaemia (16%), meningitis (15%) and endocarditis (8%). Almost 96% of the isolates could be typed using the 28 antisera. The most common serotype was serotype 2 (29%), followed by serotype 7 (17%), and serotypes 3, 4 and 8 (9-10%). The remaining serotypes were observed in frequencies of less than 5%. Serotype 7 was more commonly isolated from septicaemia than the other serotypes. Serotype 2 was more commonly isolated from pigs older than 4 weeks compared to the other serotypes. Most isolates were susceptible to amoxycillin + clavulanate, ampicillin, ceftiofur, enrofloxacin, penicillin, spectinomycin, tiamulin and trimethoprim + sulphadiazine. A high frequency (> 30%) of resistance to tetracycline was observed. Among isolates of serotype 2, 9.7% were resistant to lincomycin and 12.9% to spiramycin. Among other serotypes 56.8% were resistant to lincomycin and spiramycin. The differences in susceptibility between isolates of serotype 2 and the other serotypes were statistically significant. Compared to a previous Danish study the distribution of serotypes of S. suis causing infections among pigs in Denmark has changed during the last 15 years.

Risk factors for intestinal pathogens in Danish finishing pig herds

The objective of this investigation was to identify risk factors for infection with the intestinal bacteria: Lawsonia intracellularis, Brachyspira hyodysenteriae, Serpulina intermedia, Brachyspira innocens, Brachyspira pilosicoli and swine-pathogenic Escherichia coli (serogroups O138, O139, O141 and O149) in Danish finishing pig herds.A total of 79 herds was randomly selected and visited during 1998. From each herd, 20 faecal samples were collected from individual pigs weighing 30-50 kg. In total, 1580 faecal samples were collected and examined by polymerase chain reaction (L. intracellularis) or culture (all other agents). Information on feed and management procedures was collected by filling in questionnaires at the herd visits. The questionnaires included information on 29 dichotomous variables and three continuous variables. Variables with P<0.25 in a preliminary screening (chi2- or t-test) were selected for the statistical modelling. Our conclusions, based on the results of multifactorial logistic regression (cut-off: P=0.05), were the following: 1. Consistent batch production was associated with reduced prevalences of L. intracellularis and weakly -haemolytic spirochetes (S. intermedia, B. innocens, B. pilosicoli) (ORs=0.43 and 0.06, respectively). 2. Home-mixed (and/or non-pelleted) feed was associated with reduced prevalences of L. intracellularis and weakly -haemolytic spirochetes (ORs=0.6 and 0.4, respectively). 3. Providing straw to finishers was associated with a reduced prevalence of weakly -haemolytic spirochetes (OR=0.28-0.32). 4. Not using antimicrobial growth promoters for piglets was associated with an increased prevalence of S. intermedia (OR=11.11). 5. Rare occurrence of post-weaning diarrhoea (as opposed to common) was associated with an increased prevalence of weakly -haemolytic spirochetes (OR=8.3-13.7).

Specific Detection of Lawsonia intracellularis in Porcine Proliferative Enteropathy Inferred from Fluorescent rRNA In Situ Hybridization

Fluorescent in situ hybridization targeting 16S ribosomal RNA was used for specific detection of the obligate intracellular bacterium Lawsonia intracellularis in enterocytes from pigs affected by proliferative enteropathy. A specific oligonucleotide probe was designed and the specificity of the probe was determined by simultaneous comparison with indirect immunofluorescence assay for detection of L. intracellularis in formalin-fixed tissue samples from 15 pigs affected by porcine proliferative enteropathy. We used 10 tissue samples from pigs without proliferative mucosal changes as negative controls. The results showed that the oligonucleotide probe is specific for L. intracellularis and that fluorescent in situ hybridization targeting ribosomal RNA is a suitable and fast method for specific detection and histological recognition of L. intracellularis in formalin-fixed tissue.

The cox regression model applied to risk factor analysis of infections in the breeding and multiplying herds in the Danish SPF system

Abstract The Cox regression model is discussed with emphasis on model examination and selection. The model is applied in a multivariate risk factor analysis of Mycoplasma hyopneumoniae (MH) infections in the breeding and multiplying herds in the Danish specific pathogen-free (SPF) pig production system. The analysis showed that the pattern of infections was in agreement with the theory that both airborne transmission and spread through trade in subclinically infected animals are major causes of MH infections.

Dust Control in Pig Houses by Spraying Rapeseed Oil

Airborne dust concentrations in pig houses were reduced by spraying with a mixture of water and rapeseed oil (1 to 4 treatments d–1). The daily doses of oil (5 to 64 mL pig–1 d–1) were varied as the oil concentrations (5 to 20%) and the duration of the spraying time (5 to 90 s d–1) were changed. The treatment did not cause an increase of the average concentration of vegetable oil in the air. No pathological lung change in the pigs related to the treatment with rapeseed oil was found, and no indication of oil resorption to lung tissues, lymph nodes, or upper respiratory systems in the pigs was seen.

Identification of porcine Pneumocystis carinii as a genetically distinct organism by DNA amplification

DNA was amplified from lung samples from three piglets infected with Pneumocystis carinii, using oligonucleotide primers designed to the P. carinii mitochondrial large subunit ribosomal RNA gene. The nucleotide sequence of the amplification product was determined and indicated lack of sequence variation among these pig‐derived P. carinii samples at this locus. The data showed that porcine P. carinii was genetically distinct from P. carinii isolated from other mammalian host species.