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Colonic mucus plays a key role in colonic drug absorption. Mucus permeation assays could therefore provide useful insights and support rational formulation development in the early stages of drug development. However, the collection of native colonic mucus from animal sources is labor-intensive, does not yield amounts that allow for routine experimentation, and raises ethical concerns. In the present study, we developed an in vitro porcine artificial colonic mucus model based on the characterization of native colonic mucus. The structural properties of the artificial colonic mucus were validated against the native secretion for their ability to capture key diffusion patterns of macromolecules in native mucus. Moreover, the artificial colonic mucus could be stored under common laboratory conditions, without compromising its barrier properties. In conclusion, the porcine artificial colonic mucus model can be considered a biorelevant way to study the diffusion behavior of drug candidates in colonic mucus. It is a cost-efficient screening tool easily incorporated into the early stages of drug development and it contributes to the implementation of the 3Rs (refinement, reduction, and replacement of animals) in the drug development process.

In order to identify and evaluate health related constraints faced by Ugandan pig farmers, a veterinary herd health management approach (VHHM) was applied in 20 randomly selected pig farms in the Lira district, Uganda. Regular herd visits were conducted between July 2018 and June 2019, using e.g. interviews, observations, clinical examinations and laboratory analyzes to gather qualitative and quantitative data on relevant aspects of the production. The pig farmers kept on average 18.6 pigs, including 2.6 sows/year. The production figures varied considerably but were generally poor. The sows produced 1.6 litters/year and 8.2 piglets born alive per litter, the average daily gain was 101 g/day, and the mortality in growers was 9.7%. Four major constraints were identified; poor nutrition, infectious diseases, inferior biosecurity, and poor reproductive management. The quantity and quality of feed was suboptimal. Endo- and ectoparasites were very common, causing diarrhea, bronchitis, pneumonia, skin lesions and pruritus. Post-weaning diarrhea associated with enterotoxigenic Escherichia coli was important in the two largest herds, and parvoviral antibodies were found in seven herds, two experiencing problems with mummified fetuses. Biosecurity practices were insufficient and inconsistent, with free-ranging pigs and the use of village boars being the major risks. Reproductive figures were affected by poor estrus detection and service management. Overall, farmers lacked important knowledge on good management practices. In conclusion, the VHHM identified several important constraints that should be addressed in order to increase the productivity of Ugandan pig herds.

Over the last two decades, the pig population in Africa has grown rapidly, reflecting the increased adoption of pig production as an important economic activity. Of all species, pigs are likely to constitute a greater share of the growth in the livestock subsector. However, constraints such as respiratory infectious diseases cause significant economic losses to the pig industry worldwide. Compared to industrialized countries, the occurrence and impacts of respiratory diseases on pig production in Africa is under-documented. Hence, knowledge on prevalence and incidence of economically important swine respiratory pathogens in pigs in Africa is necessary to guide interventions for prevention and control. The purpose of this review was to document the current status of research on five important respiratory pathogens of swine in Africa to inform future research and interventions. The pathogens included were porcine reproductive and respiratory syndrome virus (PPRSv), porcine circovirus 2 (PCV2), Mycoplasma hyopneumoniae (M. hyopneumoniae), Actinobacillus pleuropneumoniae (APP) and swine influenza A viruses (IAV). For this review, published articles were obtained using Harzing’s Publish or Perish software tool from GoogleScholar. Articles were also sourced from PubMed, ScienceDirect, FAO and OIE websites. The terms used for the search were Africa, swine or porcine, respiratory pathogens, M. hyopneumoniae, APP, PCV2, PPRSv, IAV, prevention and control. In all, 146 articles found were considered relevant, and upon further screening, only 85 articles were retained for the review. The search was limited to studies published from 2000 to 2019. Of all the studies that documented occurrence of the five respiratory pathogens, most were on IAV (48.4%, n = 15), followed by PCV2 (25.8%, n = 8), PPRSv (19.4%, n = 6), while only one study (3.2%, n = 1) reported APP and M. hyopneumoniae. This review highlights knowledge and information gaps on epidemiologic aspects as well as economic impacts of the various pathogens reported in swine in Africa, which calls for further studies.

To investigate the prevalence of mastitis in sows suffering from postpartum dysgalactia syndrome (PDS), we examined milk constituents and concentrations of lipopolysaccharides (LPS) obtained from the udder vein (v. epigastrica). As part of a case–cohort study, 109 sows were monitored daily from 60 h antepartum (a.p.) to 36 h postpartum (p.p.). Over time, 38 sows were diagnosed with PDS (PDS+) and were retrospectively matched with 38 healthy sows (PDS−). The study period was divided into 7 smaller time periods (A, B, C, D, E, F, G, H, and E), allowing the studied values, in period B to G, to be compared with period A that served as a baseline, and PDS+ and PDS− sows were compared within the time periods. All sows were subjected to a thorough daily clinical examination and blood was sampled from v. epigastrica for LPS detection. Milk samples were obtained for bacteriological evaluation and detection of N-acetyl-beta-d-glucosaminidase (NAGase), lactate dehydrogenase (LDH), β-glucuronidase (β-glu; for evaluation of mastitis), isocitrate (isoC), free glucose, uric acid (UA; for evaluation of the mammary energy status), β-hydroxybutyrate acid (BHBA; for evaluation of ketosis), and milk urea (for evaluation of the protein status). The results revealed that PDS+ sows had decreased concentrations of UA in milk (P < 0.0001), increased heart rates (P < 0.01), increased mammary edema (P < 0.05), and prolonged capillary refill time in the vulvar mucosa (P < 0.01) compared with PDS− sows. Compared with baseline, feces became more solid 0 to 36 h p.p. (P < 0.0001) and the respiration rate decreased 12 to 24 h p.p. (P < 0.0001) for both PDS+ and PDS− sows. No differences were found between PDS+ and PDS− sows for severe bacterial infections, concentrations of LPS in blood or LDH, NAGase, BHBA, free glucose, isoC, or urea in milk. Concentrations of LPS in blood were not associated with signs of mastitis or edema in the mammary glands. However, a difference over time was seen for redness (P < 0.0001), warmth (P < 0.0001), and hardness (P < 0.05) of the 6 most anterior glands in both PDS+ and PDS− sows from 60 h a.p to 36 h p.p. The PDS− sows had greater concentrations of β-glu than the PDS+ sows, but no change over time was demonstrated for this marker. In conclusion, signs of mastitis were not consistently linked to PDS in sows. However, the cardiovascular system seemed to be compromised in PDS+ sows and the cause should be investigated to elucidate the pathogenesis of PDS.

Streptococcus suis is an important bacterial pathogen in pigs that may also cause zoonotic disease in humans. The aim of the study was to evaluate matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) identification of S. suis case isolates from diseased pigs and tonsil isolates from healthy pigs and wild boar using sequence analysis methods. Isolates (n = 348) that had been classified as S. suis by MALDI-TOF MS were whole-genome sequenced and investigated using analyses of (i) the 16S rRNA gene, (ii) the recN gene, and (iii) whole-genome average nucleotide identity (ANI). Analysis of the 16S rRNA gene indicated that 82.8% (288 out of 348) of the isolates were S. suis, while recN gene analysis indicated that 75.6% (263 out of 348) were S. suis. ANI analysis classified 44.3% (154 out of 348) as S. suis. In total, 44% (153 out of 348) of the investigated isolates were classified as S. suis by all of the species identification methods employed. The mean MALDI-TOF MS score was significantly higher for the S. suis case isolates than for the tonsil isolates; however, the difference is of limited practical use. The results show that species confirmation beyond MALDI-TOF MS is needed for S. suis isolates. Since the resolution of 16S rRNA gene analysis is too low for Streptococcus spp., ANI analysis with a slightly lowered cutoff of 94% may be used instead of, or in addition to, recN gene analysis. Supplementation of the MALDI-TOF MS reference library with mass spectra from S. orisratti, S. parasuis, S. ruminantium, and additional S. suis serotypes should be considered in order to produce more accurate classifications.

The global pig production has undergone major changes over the past 30 years with larger farms, more intensified production as well as improved hygiene and biosecurity practices. To investigate whether these changes, along with expanded pig welfare, have had an impact on parasite occurrence, a cross-sectional study was conducted in Sweden on farms where the pigs are always loose-housed, floors are solid and bedding material is provided. A total of 1615 faecal samples were collected on 42 conventional indoor farms from a) post-weaning piglets (n = 337); b) growers (n = 345); c) fatteners (n = 308); d) dry sows (n = 277) and e) pre-partum sows (n = 348). Samples were analysed using centrifugal flotation with a saturated glucose-salt solution and a modified McMaster technique, with a lower detection limit of 50 eggs or oocysts per gram. Samples positive for strongyle-type eggs were cultured to third stage larvae for genus identification. Farms also responded to a questionnaire regarding biosecurity, hygienic measures, and other management routines. Risk factors for parasite occurrence were assessed using mixed-effects logistical regression to account for farm-level clustering of samples. Interestingly, the prevalence of Ascaris suum was reduced compared to a similar investigation in the 1980s. In the present study A. suum was detected only in 43 % of the herds, with the highest prevalence in pre-partum sows (37 %) followed by fatteners (25 %). Small sized farms were associated with higher odds of being positive, compared to large sized farms (OR = 159.1, P = 0.010). Oesophagostomum spp. were detected in 64 % of the herds and again mainly in pre-partum sows (63 %). Trichuris suis was detected in 10 % of the herds but only in

The gastrointestinal mucus is a hydrogel that lines the luminal side of the gastrointestinal epithelium, offering barrier protection from pathogens and lubrication of the intraluminal contents. These barrier properties likewise affect nutrients and drugs that need to penetrate the mucus to reach the epithelium prior to absorption. In order to assess the potential impact of the mucus on drug absorption, we need information about the nature of the gastrointestinal mucus. Today, most of the relevant available literature is mainly derived from rodent studies. In this work, we used a larger animal species, the pig model, to characterize the mucus throughout the length of the gastrointestinal tract. This is the first report of the physiological properties (physical appearance, pH and water content), composition (protein, lipid and metabolite content) and structural profiling (rheology and gel network) of the porcine gastrointestinal mucus. These findings allow for direct comparisons between the characteristics of mucus from various segments and can be further utilized to improve our understanding of the role of the mucus on region dependent drug absorption. Additionally, the present work is expected to contribute to the assessment of the porcine model as a preclinical species in the drug development process.

Lawsonia intracellularis is a Gram-negative obligate intracellular bacterium that is the aetiological agent of proliferative enteropathy (PE), a common intestinal disease of major economic importance in pigs and other animal species. To date, progress in understanding the biology of L. intracellularis for improved disease control has been hampered by the inability to culture the organism in vitro. In particular, our understanding of the genomic diversity and population structure of clinical L. intercellularis is very limited. Here, we utilized a metagenomic shotgun approach to directly sequence and assemble 21 L . intracellularis genomes from faecal and ileum samples of infected pigs and horses across three continents. Phylogenetic analysis revealed a genetically monomorphic clonal lineage responsible for infections in pigs, with distinct subtypes associated with infections in horses. The genome was highly conserved, with 94 % of genes shared by all isolates and a very small accessory genome made up of only 84 genes across all sequenced strains. In part, the accessory genome was represented by regions with a high density of SNPs, indicative of recombination events importing novel gene alleles. In summary, our analysis provides the first view of the population structure for L. intracellularis , revealing a single major lineage associated with disease of pigs. The limited diversity and broad geographical distribution suggest the recent emergence and clonal expansion of an important livestock pathogen.

Lawsonia intracellularis is a Gram-negative obligate intracellular bacterium that is the aetiological agent of proliferative enteropathy (PE), a common intestinal disease of major economic importance in pigs and other animal species. To date, progress in understanding the biology of L. intracellularis for improved disease control has been hampered by the inability to culture the organism in vitro. In particular, our understanding of the genomic diversity and population structure of clinical L. intercellularis is very limited. Here, we utilized a metagenomic shotgun approach to directly sequence and assemble 21 L. intracellularis genomes from faecal and ileum samples of infected pigs and horses across three continents. Phylogenetic analysis revealed a genetically monomorphic clonal lineage responsible for infections in pigs, with distinct subtypes associated with infections in horses. The genome was highly conserved, with 94 % of genes shared by all isolates and a very small accessory genome made up of only 84 genes across all sequenced strains. In part, the accessory genome was represented by regions with a high density of SNPs, indicative of recombination events importing novel gene alleles. In summary, our analysis provides the first view of the population structure for L. intracellularis, revealing a single major lineage associated with disease of pigs. The limited diversity and broad geographical distribution suggest the recent emergence and clonal expansion of an important livestock pathogen. ispartof: MICROBIAL GENOMICS vol:6 issue:4 ispartof: location:England status: published

Salmonellosis and yersiniosis are notifiable human diseases that are commonly associated with contaminated food. Domestic pigs as well as wild boars and other wild-life have been identified as reservoirs of these bacteria. Methods for cultivation and molecular epidemiological investigations of Salmonella spp. are well established, however, cultivation of enteropathogenic Yersinia spp. is time- consuming and the commonly used method for molecular epidemiological investigations, pulsed-field gel electrophoresis, lack in discriminatory power. The aim of this study was to develop and evaluate a screening protocol well suited for wildlife samples and other highly contaminated samples. The method is based on PCR-screening followed by Multiple Loci Variant number tandem repeat Analysis (MLVA) on enrichment broth to obtain molecular epidemiological data for enteropathogenic Yersinia spp. without the need for pure isolates. The performance of the protocol was evaluated using wild boar samples (n=354) including tonsils, faeces and lymph nodes from 90 Swedish wild boars. The new protocol performed as well as or better than the established ISO-standards for detection and cultivation of Y. enterocolitica and Salmonella spp., however for cultivation of Y. pseudotuberculosis, further development is needed. The selection for motility seems beneficial for the enrichment of Salmonella spp. and Y. enterocolitica. Further, the selective enrichment prior to PCR-analysis eliminates inhibitory factors present in the original sample. In total, ten isolates of Y. enterocolitica of various bio-serotypes were obtained, and the MLVA-profile of these isolates were consistent with the profiles from the corresponding enrichment broth. Further, 22 isolates of Salmonella spp. comprising six different serovars were obtained with S. Fulica, S. Hadar and a monophasic S. Typhimurium being the most common. In conclusion, the presented screening protocol offers a rapid and efficient way to obtain prevalence data from a large sample set as well as MLVA-data within a short time frame. These results can hence improve the knowledge on the epidemiology and distribution of these pathogens and their importance to public health.

Congenital tremor (CT) is a neurological disease that affects new-born piglets. It was described in 1922 and six different forms, designated type AI-V and type B, are described based on the causative agents, as well as specific histological findings in the central nervous system (CNS). The various forms present with identical clinical signs consisting of mild to severe tremor of the head and body, sometimes complicated with ataxia. By definition, all A-forms have hypomyelination of the CNS, whereas there are no histopathological lesions with the B-form. The cause of the A-II form was long unknown, however, at present several different viruses have been proposed as the causative agent: porcine circovirus-II (PCV-II), astrovirus, PCV-like virus P1, and atypical porcine pestivirus (APPV). Currently, APPV is the only virus that has been proven to fulfill Mokili's Metagenomic Koch's Postulates. Following infection of the pregnant sow, the virus passes the placental barrier and infects the fetus. Interestingly, no clinical signs of disease have been associated with APPV in adult pigs. Furthermore, other viruses cannot be ruled out as additional potential causes of CT. Given the increased interest and research in CT type A-II, the aim of this review is to summarize current knowledge.

The original article [1] contains an error whereby the caption in Figure 8 is incorrect; the correct caption can be seen ahead alongside its respective image.

SUMMARYThe European wild boar populations are growing and spreading to new areas, which might constitute a threat to public health, since wild boar can harbour pathogens with the potential to cause serious illness in humans. Tonsils, ileocaecal lymph nodes and faecal samples were collected from 88 Swedish wild boars and analysed for the presence of the zoonotic pathogensSalmonellaspp.,Yersinia enterocolitica, Y. pseudotuberculosisand enterohaemorrhagicEscherichia coliO157:H7 (EHEC). A combination of cultivation and polymerase chain reaction (PCR) analysis was used and overall, 20% of sampled individuals tested positive forY. enterocolitica, 20% forY. pseudotuberculosisand 10% forSalmonellaspp. A total of 41% of sampled individuals tested positive for one or more of these three pathogens. No EHEC were detected. Samples PCR-positive forSalmonellaspp. were cultivated further and six isolates were obtained, belonging toSalmonella entericasubspeciesentericaand subspeciesdiarizone. The pathogens were most commonly detected in tonsil samples.

SUMMARYA total of 207 wild rodents were caught on nine pig farms, five chicken farms and five non-farm locations in Sweden and surveyed for a selection of bacteria, parasites and viruses.Lawsonia intracellulariaand pathogenicYersinia enterocoliticawere only detected in rodents on pig farms (9% and 8% prevalence, respectively) which indicate that these agents are more likely to be transmitted to rodents from pigs or the environment on infected farms.Brachyspira hyodysenteriae(1%),Brachyspira intermedia(2%),Campylobacter jejuni(4%),Campylobacter upsaliensis(2%), leptospires (7%) and encephalomyocarditis virus (9%) were also detected from rodents not in contact with farm animals.GiardiaandCryptosporidiumspp. were common, although no zoonotic types were verified, andSalmonella entericawas isolated from 1/11 mice on one farm but not detected by PCR from any of the rodents.Trichinellaspp. andToxoplasma gondiiwere not detected.

In modern pig production, proliferative enteropathy is a common cause of diarrhoea and poor growth in young animals. This study aimed to determine the possible spread of Lawsonia intracellularis through the sale of replacement gilts and the possibility to protect the herds by adequate biosecurity measures. This was achieved by repeated sampling of 50 gilts in an infected multiplying herd, from the last day in the farrowing pen and until sale. Further, 60 gilts sold from this herd were tested during their stay in quarantine in a recipient herd. To confirm freedom from infection, 100 growing pigs in the recipient herd were also tested. Individual faecal ( n = 748) and blood ( n = 728) samples were analysed by PCR and ELISA, respectively. Transmission of L. intracellularis from the sows to their offspring was not demonstrated. However, the possible transmission between herds by replacement gilts was demonstrated. Peak shedding occurred at 12 and 15 weeks of age, and single animals were also PCR-positive at 24–36 weeks of age in the multiplying herd and in the quarantine in the recipient herd. Further, the possible occurrence of chronically infected carrier animals was suggested. Although L. intracellularis is widely spread, it appears possible to avoid the transmission between herds by employing adequate biosecurity measures. Thus, it would be advisable to establish herd profiles in breeding herds to avoid the selling of infected animals as well as to establish the health status of the recipient herd. Further, the health status of the recipient herds should be known.

Neonatal porcine diarrhoea of uncertain aetiology has been reported from a number of countries. This study investigated 50 diarrhoeic and 19 healthy piglets from 10 affected Swedish herds. The piglets were blood-sampled for analysis of serum γ-globulin and necropsied, and the intestines were sampled for histopathology and cultured for Escherichia coli, Clostridium perfringens and Clostridium difficile. Escherichia coli isolates (n = 276) were examined by PCR for virulence genes encoding LT, STa, STb, EAST1, VT2e, F4, F5, F6, F18, F41, AIDA-I, intimin, and for the genes aaiC and aggR. Selected isolates were analysed for additional virulence genes by a microarray and subjected to O-typing. Clostridium perfringens isolates (n = 152) were examined by PCR for genes encoding major toxins, enterotoxin and beta2-toxin. There was no difference in serum γ-globulin concentration between diarrhoeic and non-diarrhoeic piglets, and pathological lesions in the intestines were generally mild. Porcine enterotoxigenic Escherichia coli, a common cause of piglet diarrhoea, was only found in two piglets. Further, the virulence gene profiling did not suggest involvement of other diarrhoeogenic pathotypes of Escherichia coli. Growth of Clostridium perfringens did not differ between diarrhoeic and non-diarrhoeic piglets. All isolates were type A, all were negative for enterotoxin, and 151 of 152 isolates were beta2-toxin positive. In pigs ≥ 2 days old, moderate to profuse growth of Clostridium difficile was more common in the controls. In conclusion, it was not possible to relate Escherichia coli, Clostridium perfringens type A and C or Clostridium difficile to neonatal porcine diarrhoea in any of the investigated herds.

Background: Biosecurity routines at herd level may reduce the probability of introduction of disease into the herd, but some measures may be regarded as expensive and cumbersome for the farmers. Custom-made measures based on individual farm characteristics may aid in improving the actual application of on-farm biosecurity. The aim of the study was to provide a tool for calculating the effects of different biosecurity measures and strategies on the individual farm level.A simple model was developed to assess the risk of disease introduction and the need for biosecurity measures in individual farms. To illustrate the general applicability of the tool, it was applied to theoretical examples of Swedish cattle and pig farms and diseases endemic in those animal species in the EU, in two scenarios with different between-farm contact patterns.Results: The model illustrated that the most important factors affecting the risk, and the effect of biosecurity measures such as quarantine routines and protective clothing, were the frequency of between- farm contacts and prevalence of the disease. The risk of introduction as well as the effect of biosecurity measures differed between farm types and disease transmission routes. Adapting contact patterns to mitigate a specific disease risk was as important as biosecurity measures for some farm types, but the largest effect was seen when combining biosecurity measures with more planned contact patterns.Conclusions: The risk assessment model proved useful for illustrating the risk of introduction of endemic diseases and the mitigating effect of different biosecurity measures on farm level. Model outputs could be used to justify prioritisation of measures or adapting contact patterns. The theoretic exercise of adjusting model inputs and comparing outputs may help veterinary advisors to understand farm-specific risks and motivate farmers to improve biosecurity in their individual farm, as it can be tailored to each farmer's needs and preferences.

Objective To evaluate the effects of preoperative extradural morphine on the end-tidal isoflurane (F e ′ISO) concentration and on physiological variables in pigs undergoing abdominal surgery. Study design Prospective, randomized, blinded study. Animals Fourteen healthy pigs (20 ± 4 kg) undergoing intestinal cannulation. Materials and methods Anaesthesia was induced with a combination of medetomidine (50 μg kg −1 ) and tiletamine-zolazepam (2.5 mg kg −1 ) injected intramuscularly, and was maintained with isoflurane in air and oxygen (F i O 2 = 50% O 2 ). In the first group, morphine (0.1 mg kg −1 ) was administered extradurally before surgery. The second group received an equivalent volume of extradural saline as control. During the experiment, heart and respiratory rates, mean arterial blood pressure, tidal volume and minute ventilation were recorded every 10 minutes. The concentration of F e ′ISO was adjusted, according to the depth of anaesthesia, by an experienced animal nurse. Within treatment groups, time-related changes in F e ′ISO and physiological variables were analysed using a repeated measurement anova . Differences in data between treatment groups were analysed at specific time points using a Mann–Whitney U -test. Results are presented as mean ± SD; p Results After the onset of action of the morphine, the F e ′ISO required to maintain anaesthesia was significantly lower in the extradural morphine group compared with control. During the expected maximal effect of the drug, F e ′ISO was significantly lower in the morphine group (0.6 ± 0.2%) than in the control group (0.9 ± 0.2%). The decrease in F e ′ISO indicated that the onset of action of morphine was approximately 30 minutes after injection. No significant differences in other clinical variables were found between the groups. Conclusion Pigs that received extradural morphine before abdominal surgery achieved surgical anaesthetic depth at a lower F e ′ISO concentration. Clinical relevance Extradural morphine allows abdominal surgery to be performed at a lower F e ′ISO concentrations.

The objective of this work was to evaluate the physiological and behavioural effects of opioid analgesic treatment in pigs subjected to abdominal surgery. Ten Swedish Landrace x Yorkshire pigs (20 +/- 4 kg b.w.) were submitted for intestinal cannulation. The pigs were allocated into two groups during one preoperative, one surgical and two postoperative days. All pigs were anaesthetized with medetomidine, tiletamine and zolazepam. One group was treated with epidural morphine (0.1 mg/kg) preoperatively, and transdermal fentanyl patches (50 microg/kg/h) were applied behind the ear immediately after surgery. The other group received epidural saline (equivalent volume) and placebo patches. All pigs were regularly weighed and clinically examined and repeated blood samples were analysed for serum concentrations of cortisol, beta-endorphin and fentanyl. Pre- and postoperative behaviours were evaluated by a swine specialist blinded to the treatment, three times a day, and were also videotape recorded for a total of 84 h per pig. No differences in behaviour were noted by the observer. During the first postoperative 12 h, treated pigs did not differ in activity compared with preoperative recordings, while untreated pigs were found to be less active. The treated group started to show interest in eating immediately after anaesthesia recovery, whereas the placebo group did not. During the 12-60 h postoperative period, the treated group had lower activity levels compared with the preoperative levels, which were similar to those in the placebo group. Treated pigs gained 0.5 +/- 0.2 kg during the subsequent two postoperative days, whereas the untreated pigs lost weight throughout the experiment. Cortisol concentration differed immediately after the surgery: Group P had 325 +/- 120 nmol/L and Group M 159 +/- 49 nmol/L. beta-endorphin concentration did not differ between groups. The highest serum fentanyl concentration (0.37 +/- 0.3 ng/mL) was measured 24 h postoperatively. Preoperative epidural morphine in combination with postoperative transdermal fentanyl resulted in earlier return to normal activity levels and an immediate weight gain after surgery.

The aim of this study was to examine the levels of circulating leukocytes and lymphocyte subpopulations before and immediately after experimentally induced swine dysentery. Twenty-one healthy crossbred pigs (approximately 22 kg) were orally inoculated with Brachyspira hyodysenteriae. Blood was sampled before inoculation and when clinical signs of swine dysentery occurred. Pigs that remained healthy were sampled when killed. Total and differential white blood cell counts were performed, and lymphocyte subpopulations were analysed using flow cytometry. Following a mean incubation period of 13 days, 12 pigs developed swine dysentery, whereas nine remained healthy throughout the study. Before inoculation, pigs that subsequently developed swine dysentery displayed higher levels of circulating gamma delta T cells (mean +/- se; 30.7 +/- 3.5 %) compared with pigs that remained healthy (14.9 +/- 1.4 %). Sick animals also displayed lower levels of CD8 cells (24.6 +/- 1.5 %), cytotoxic/suppressor T cells (10.9 +/- 1.3 %) and CD4 CD8 T cells (8.1 +/- 1.0 %) than the pigs that remained healthy (34.9 +/- 3.1 %; 17.6 +/- 2.0 %; 13.6 +/- 2.3 %). No difference was observed in leukocyte counts before inoculation. At onset of swine dysentery, there was an increase in monocytes (from 1.5 +/- 0.2 x 10 to 3.8 +/- 0.5 x 10 l) and CD4 CD8 T cells (from 5.8 +/- 0.9 to 8.9 +/- 0.7 %). In conclusion, gamma delta T cells and CD8 cells may be associated with susceptibility to experimentally induced swine dysentery, whereas monocytes and CD4 CD8 T cells appear to be the major responding leukocytes during the disease.

Diarrhoea among growing pigs (8-13 weeks old) is a significant problem in many herds. Nine herds with poor performance and diarrhoea among growing pigs were selected on the basis of their piglet mean age at a body weight of 25 kg, compared to the overall mean age in Swedish herds. In addition, four herds with good average performance and no problems with diarrhoea were selected. Pigs were necropsied and samples for histology and microbiology were collected. Based on the necropsy findings, the pigs from the good performing herds were all judged to be healthy. The presence of Brachyspira pilosicoli and Lawsonia intracellularis was significantly correlated to poor performing herds and the results indicate that these microbes are main pathogens involved in enteric diseases among Swedish grower pigs. In addition, concomitant infections with other presumptive pathogens were commonly found.

Attempts to eliminate Sarcoptes scabiei var. suis were made in 2 piglet producing herds by intramuscular injections with doramectin (Dectomax™, Pfizer inc., New York, USA). No hygienic measures of the environments were undertaken. In herd A, all animals were treated twice at a 14-day interval. Theoretically it should be possible to eliminate the mite with a single injection and therefore the animals in herd B were treated only once. The results were measured by attempts to demonstrate S. scabiei by skin scrapings, by recording skin lesions, by establishing rubbing indexes, and by measuring serum antibodies to S. scabiei. All adult animals greater than 8 months of age and 15 weaned piglets 8–16 weeks of age per herd were investigated prior to treatments and every fourth month for a total of 20 months following administration of the treatments. Live S. scabiei were demonstrated prior to treatments in both herds, but no further. Skin lesion scores, rubbing indexes and serum antibody levels remained low throughout the study. It is possible to eliminate S. scabiei from a herd with one single injection of doramectin. Precautions must be taken to ensure that all animals get a correct dose and that the drug is properly administered.

Background Within the genera Chlamydia, the development of refined diagnostic techniques has allowed the identification of four species that are capable of infecting pigs. The epidemiology, clinical, and zoonotic impacts of these species are however largely unknown. The study aimed to investigate the presence of Chlamydia spp. in the intestines of growing pigs and in conjunctival swabs from finisher pigs, and relate the findings to clinical signs. Results By histology, 20 of 48 pigs had intestinal lesions that may be consistent with chlamydial infection. By PCR, forty-six of the pigs were positive whereas two samples were inhibited. Sequencing of 19 DNA extracts identified these as Chlamydia suis. By immunohistochemistry, 32 of 44 samples were positive and a significant relationship was detected between macroscopically visible intestinal lesions and a high degree of infection. By real-time PCR, a significant difference was detected between pigs with and without conjunctivitis when a Ct value of 36 was employed but not when a Ct value of 38 was employed. Conclusions Chlamydia suis was demonstrated in most samples and overall, no correlation to clinical signs was detected. However, a correlation was noted between samples with a high degree of infection and the presence of clinical signs. It is possible, that the intensive pig production systems studied might predispose for the transmission and maintenance of the infection thus increasing the infectious load and the risk for disease in the pig.

Background Lawsonia intracellularis is a common cause of chronic diarrhoea and poor performance in young growing pigs. Diagnosis of this obligate intracellular bacterium is based on the demonstration of the microbe or microbial DNA in tissue specimens or faecal samples, or the demonstration of L. intracellularis-specific antibodies in sera. The aim of the present study was to evaluate a blocking ELISA in the detection of serum antibodies to L. intracellularis, by comparison to the previously widely used immunofluorescent antibody test (IFAT). Methods Sera were collected from 176 pigs aged 8-12 weeks originating from 24 herds with or without problems with diarrhoea and poor performance in young growing pigs. Sera were analyzed by the blocking ELISA and by IFAT. Bayesian modelling techniques were used to account for the absence of a gold standard test and the results of the blocking ELISA was modelled against the IFAT test with a "2 dependent tests, 2 populations, no gold standard" model. Results At the finally selected cut-off value of percent inhibition (PI) 35, the diagnostic sensitivity of the blocking ELISA was 72% and the diagnostic specificity was 93%. The positive predictive value was 0.82 and the negative predictive value was 0.89, at the observed prevalence of 33.5%. Conclusion The sensitivity and specificity as evaluated by Bayesian statistic techniques differed from that previously reported. Properties of diagnostic tests may well vary between countries, laboratories and among populations of animals. In the absence of a true gold standard, the importance of validating new methods by appropriate statistical methods and with respect to the target population must be emphasized.

This field study explored the cytokine expression in intestinal tissue and serum from 19 diarrhoeic and 9 healthy pigs in herds with a long-time history of Lawsonia intracellularis -infection. The disease, proliferative enteropathy (PE), is associated with diarrhoea and poor performance in growers and haemorrhagic diarrhoea and sudden death in finisher pigs, but the immunopathology is poorly understood. Histopathology, demonstration of L. intracellularis and porcine circovirus type 2 (PCV2) in intestinal tissue by PCR, and detection of serum antibodies to L. intracellularis, were performed. The presence of IL-1β, IL-6, IL-10, IFN-α, IFN-γ, TNF-α and TGF-β in sera was determined by immunoassays, and intestinal mRNA expression of these cytokines plus IL-12p40 was determined by qPCR. Intestinal specimens from pigs with intestinal adenomatosis ( n = 2), proliferative haemorrhagic enteropathy or swine dysentery ( n = 2), and controls ( n = 2) were analysed by a genome wide porcine microarray. The clinical signs of PE were not always supported by the subsequent analyses, and the presence of PCV2 may have contributed to an increased mRNA expression for IFN-γ in intestinal specimens from some pigs. The limited gene expression in the microarray analyses and the limited expression of cytokines in both sera and intestines, indicate that the immune response is poorly activated in the initial course of an infection with L. intracellularis . However, the gene encoding for insulin-like growth factor binding protein 3 (IGFBP-3) was up-regulated in two pigs with prominent mucosal proliferation.

The obligate intracellular bacteriumLawsonia intracellulariscauses enteritis and poor growth in weaned pigs. Cultivation is difficult and diagnosisante mortemis mainly based on techniques such as polymerase chain reaction. However, false negative results caused by the presence of PCR-inhibitory factors constitute a problem. This study aimed to develop and evaluate a new technique, flotation, to separateL. intracellularisfrom inhibitors in faeces prior to PCR. The technique was evaluated by comparison to two previously evaluated and commonly used methods, preparation by boiling lysate combined with nested PCR and preparation by a commercial kit combined with conventional PCR. Continuous density centrifugation of faecal samples containingL. intracellularissuggested the buoyant density of the microbe to be between 1.064 and 1.077 g/mL. Several flotation setups were tested to achieve optimal separation of the microbe from inhibitors and faecal particles. The finally selected setup floated wholeL. intracellularisfrom the application site at the bottom to the upper part of the gradient while inhibitory components mainly remained in the bottom. PCR was performed directly on material recovered from the upper interphase. The method was evaluated on 116 clinical samples. As compared to sample preparation by boiling combined with nested PCR, fewer samples were inhibited but also fewer positives were identified. In comparison to preparation by a commercial kit combined with conventional PCR, presently used for routine diagnosis, similar results were obtained. However, the new method was comparably faster to perform. The new method, based on flotation ofLawsonia intracellulariscombined with conventional PCR, was well suited for routine diagnosis.

Proliferative enteropathy caused by the intracellular bacterium Lawsonia intracellularis is an endemic disease with high herd prevalences reported worldwide. The infection has a considerable impact on pig production and herd economics and, with the development of new diagnostic techniques, L. intracellularis is being identified in an increasing number of pig herds and a wider range of species. This paper reviews current knowledge of the disease, with a focus on the epidemiology in pigs. The prevalence of infection, transmission, predisposing factors, microbial features, pathogenesis, clinical signs, diagnosis, treatment and control are discussed. The disease is mainly controlled by antibiotic treatment and vaccination at herd level. In the development of effective measures to prevent the spread of the infection, increased knowledge of the transmission and persistence of the microorganism are crucial.

Summary A traditional design for studies of pathogenesis of experimentally induced diseases of the large intestine in individual pigs involves euthanasia and necropsy at scheduled times. An alternative design has been developed enabling the sequential in vivo monitoring of events in the intestine of the individual pig before and during disease. The alternative design is based on repeated endoscopy and biopsy sampling to monitor the course of the disease. The purpose of this study was to compare the statistical properties of the two designs. One hundred replicates of a computer-generated simulation of data were performed. In the simulation, data were generated for individual pigs according to the alternative design, i.e. four sequential observations on the outcome of interest, and the ‘traditional’ design was created by retaining only one observation per pig. Data from the alternative design were analysed with a generalized linear mixed model, accounting for the clustering of observations within animals and the time-dependence between observations within animals, i.e. observations within animals are interdependent and follow a pattern over time. Data from the ‘traditional’ design were analysed with an ordinary linear model. The results showed that the alternative design was much more efficient than the traditional and that only one-fourth as many pigs were required to achieve the same precision in estimated effects.

Summary The aim of the present study was to survey the prevalences of the enteric pathogens Brachyspira hyodysenteriae, Brachyspira pilosicoli and Lawsonia intracellularis in Swedish growing pigs and in the Swedish wild boar population and to relate these findings to clinical signs. The study included 105 randomly selected herds, constituting approximately one third of Swedish herds with a herd size of >100 sows. The herds were located all over the country. In these herds, growth promoters were not used and pigs sampled were not subjected to any medication. From each herd, samples were taken from 10 growing pigs aged 8–12 weeks, corresponding to approximately 2.5% of all growing pigs present in the herd at the sampling occasion. If possible, the samples were taken from pigs with diarrhoea. Forty-eight faecal samples and 71 rectal swabs were also taken from free-living wild boars (31 piglets, 19 growers and 21 adult animals) at shooting. The samples were analysed by culture and biochemical tests for the presence of Brachyspira spp. and by nested PCR for the presence of L. intracellularis. Brachyspira hyodysenteriae was not demonstrated in any sample. Brachyspira intermedia was detected in 22 samples originating from 15 herds, Brachyspira innocens/ Brachyspira murdochii was detected in 370 samples from 82 herds and B. pilosicoli was detected in 134 samples originating from 34 herds. In 21 herds and in 534 samples, no Brachyspira spp. were detected. Lawsonia intracellularis was demonstrated in 285 samples from 50 herds. Further, 418 samples from conventional herds were negative with respect to L. intracellularis and in 345 samples the PCR had been inhibited. All samples from the wild boars were negative for Brachyspira spp., 12 of 48 samples were negative for L. intracellularis, and in 36 wild boar samples, the PCR was inhibited.

The development of intestinal lesions after inoculation with Brachyspira hyodysenteriae was followed by repeated endoscopy and biopsy sampling through a caecal cannula. Seven eight-week-old pigs were cannulated and inoculated, two were cannulated but not inoculated, and two pigs were inoculated but not cannulated. Endoscopy, biopsy, and blood sampling to determine SAA (serum amyloid A), haptoglobin, cortisol, and WBC counts were performed at scheduled time-points. At the third day of disease, endoscopy showed a hyperaemic, perturbed mucosa and excessive amount of mucus. Histologically, crypt hyperplasia, depletion of goblet cell mucus, and erosions were noted. Simultaneously, elevated acute phase proteins and circulating monocytes, and decreased number of intraepithelial CD3(+) cells were observed. After five days the pigs recovered. Intestinal lesions were demarcated and interspersed among apparently normal mucosa and blood parameters returned to initial values. Endoscopy through an intestinal cannula made it possible to follow the development of intestinal alterations in vivo and describe the sequential events during the course of swine dysentery. The number of animals used in a study could thus be minimised and the precision of the experiment increased.

The aim of this study was to find suitable and reliable tools for demonstrating Lawsonia intracellularis in routine clinical diagnosis. Firstly, a method to prepare tissue samples before a polymerase chain reaction (PCR) was evaluated in pigs submitted for necropsy. Secondly, seven different faecal preparation methods and four different DNA polymerases were tested in single or nested PCR, with co-amplification of a mimic molecule. Thirdly, in selected pigs submitted for necropsy, tissue and faecal samples were examined histopathologically and by PCR, and blood samples were analysed serologically. Detection of L. intracellularis in tissue preparations by PCR showed good specificity and correlated to lesions found at necropsy. The sensitivity in spiked tissue samples was 10(1)-10(2) mimic molecules per tube. In faecal samples, nested PCR on boiled lysate gave the best result with a sensitivity of 10(2)-10(3) mimic molecules per reaction tube. However, because of the time-consuming procedure and the increased risk for contamination, a commercially available kit was preferred for routine diagnoses, despite a somewhat lower detection rate in subclinically infected pigs. In a few cases, the serological results differed from those obtained by PCR and by necropsy but the reason for this is not clear. This study indicates that the best method for diagnosis of acute enteritis in growers is PCR on faecal or tissue samples. To determine the presence of the bacteria in a herd, serology or repeated faecal sampling for PCR from target animals, or both, should be used.

Lawsonia intracellularis is an intracellular organism that causes proliferative enteritis in pigs. This bacterium is difficult to culture, and antemortem demonstration of the microbe is therefore often performed on fecal samples by polymerase chain reaction (PCR). Polymerase chain reaction is sensitive and specific, but inhibitory factors in feces might cause false-negative results. This article describes the construction and use of an internal standard, a mimic. The mimic is amplified by the same primers as those used for L. intracellularis DNA and thus could indicate false-negative results in clinical samples. The amplicon was clearly visible when as few as 10 mimic molecules were added per amplification reaction and when no inhibitors were present. When fecal samples were spiked with the mimic, the detection limit was 10 2 molecules per PCR. Sixty clinical samples, 20 from wild boars, 20 from growing pigs with diarrhea, and 20 from pigs without diarrhea, were prepared by a boiling procedure and subjected to PCR together with 10 3 mimic molecules. Nine samples were positive, of which 7 originated from pigs with diarrhea and 2 from pigs without diarrhea. In 14 samples from wild boars, in 8 samples from pigs without diarrhea, and in 3 samples from pigs with diarrhea, neither the mimic nor the target DNA was visible. This indicated the presence of inhibitors in these samples. It is concluded that the mimic can be used as an internal control in the diagnosis of L. intracellularis to indicate inhibition of PCR. ples. This phenomenon may be caused by a poor re- covery rate in the sample preparation method or by the presence of various PCR-inhibitory factors in the feces. 2,3,10,19 A method to detect PCR inhibition using an internal standard has been previously described. 17 The technique is based on a DNA molecule, a mimic, that differs in sequence from the target template but uses the same primer recognition sites, which allow it to mimic the template through the PCR amplification. The mimic amplicon differs in size from the target amplicon and could thus be separated and distin- guished by gel electrophoresis. The technique has been further developed for detecting some viral and bacte- rial diseases. 1,5 The aim of the present study was to develop an internal standard, a mimic, that could be used to dem- onstrate the presence of PCR-inhibitory factors in fecal samples subjected to analysis for L. intracellularis. This would facilitate the interpretation of negative re- sults and thus further strengthen the reliability of the diagnostic method.

Eradication programmes directed against Sarcoptes scabiei var. suis, performed without any disinfection of the environment, were compared to an eradication programme including a simultaneous environmental disinfection. The performance of such programmes would be facilitated considerably if environmental disinfection could be avoided. The trial was conducted in four conventional piglet-producing herds designated A-D. All herds suffered from a subclinical but persistent infection with the sarcoptic mite. Herd A was treated with two injections of ivermectin and simultaneously disinfection of the environment was done. In the other herds, the animals were treated twice (B and C) or three times (D) with acaricides (ivermectin or phoxim) at intervals of 7-12 days without any simultaneous disinfection of the buildings or environment. The results were monitored by attempts to demonstrate the mite by skin scrapings, and by monitoring the levels of serum antibodies to Sarcoptes scabiei by an ELISA. Samples were taken from all adults and from 20% of the piglets aged 10-12 weeks, before initiating the control programmes, every third month during the first year and a final sampling 18 months after the treatments. After treatment, the levels of antibodies continuously decreased. Presumably dead S. scabiei were demonstrated in one sow from two of the herds 3 months after initiating acaricidal treatments. S. scabiei was not demonstrated later, nor from skin scrapings of the piglets. The results obtained demonstrate that it is possible to eradicate sarcoptic mange mites in swine herds with the use of simplified eradication programmes. The success of such programmes may efficiently be monitored by measuring the levels of serum antibodies to S. scabiei.