Influence of diet on the experimental infection of pigs with Brachyspira pilosicoli

The effects of five different diets on the experimental infection of pigs with a Danish field isolate of Brachyspira pilosicoli were investigated. The diets tested were a pelleted and a non-pelleted standard diet based on wheat and barley, the standard diet supplemented with 2 per cent lactic acid, a fermented liquid feed and a diet based on cooked rice. Two trials were conducted, each with six groups of six pigs; in each, two of the groups were fed the standard diet. One of these groups and the other four groups were challenged after two weeks on the diets and euthanased four weeks later. The clinical signs of B pilosicoli infection varied from loose stools to watery, mucoid diarrhoea. The group fed the rice diet excreted B pilosicoli in their faeces for a significantly shorter period than the group fed the standard diet (P < 0.01), and fewer of them excreted the organism (P < 0.05). All the pigs fed the pelleted diet excreted B pilosicoli in their faeces, and significantly more of them showed clinical signs of disease than the pigs fed the standard diet (P < 0.05). The fermented liquid feed and the diet containing lactic acid had no significant effect on the excretion of B pilosicoli or on the numbers of pigs showing clinical signs of disease.     

Brachyspira pilosicoli isolated from pigs in Japan

Two of four weak beta-hemolytic isolates of intestinal spirochetes isolated from pigs in Japan possessed a unique base alignment of TTTTTT on the 16S ribosomal DNA of Brachyspira pilosicoli and were identified as B. pilosicoli. The other two isolates were not identified by this technique. The identified isolates were 4.2 to 11 microm in length and 0.2 to 0.3 microm in diameter, 4 periplasmic flagella at each end were observed dominantly. The isolates were hippurate positive but indole negative. This is the first report on the isolation of B. pilosicoli from pigs in Japan.     

Cecal spirochetosis caused by Brachyspira pilosicoli in commercial turkeys

Spirochetes that were identified as Brachyspira pilosicoli were present in the ceca of 7.5- to 18-wk-old turkeys with cecal spirochetosis and typhlitis. The identity of B. pilosicoli was confirmed on the basis of ultrastructural morphology of the cecal epithelium adherent microbes, immunohistochemical staining with a Brachyspira genus-specific monoclonal antibody, and amplification of a B. pilosicoli species-specific 16S ribosomal RNA (rrs gene) sequence by using the polymerase chain reaction and DNA obtained by laser-capture microdissection of the epithelium-adherent microbial fringe. To the author's knowledge, this is the first report of B. pilosicoli in the ceca of turkeys.     

Detection of Brachyspira hyodysenteriae, Lawsonia intracellularis and Brachyspira pilosicoli in feral pigs

Feral pigs are recognized as being a potential reservoir of pathogenic microorganisms that can infect domestic pigs and other species. The aim of this study was to investigate whether feral pigs in Western Australia were colonized by the pathogenic enteric bacteria Lawsonia intracellularis, Brachyspira hyodysenteriae and/or Brachyspira pilosicoli. A total of 222 feral pigs from three study-populations were sampled. DNA was extracted from faeces or colonic contents and subjected to a previously described multiplex PCR for the three pathogenic bacterial species. A subset of 61 samples was cultured for Brachyspira species. A total of 42 (18.9%) of the 222 samples were PCR positive for L. intracellularis, 18 (8.1%) for B. hyodysenteriae and 1 (0.45%) for B. pilosicoli. Four samples were positive for both L. intracellularis and B. hyodysenteriae. Samples positive for the latter two pathogens were found in pigs from all three study-sites. A strongly haemolytic B. hyodysenteriae isolate was recovered from one of the 61 cultured samples. Comparison of a 1250-base pair region of the 16S rRNA gene amplified from DNA extracted from the isolate and five of the B. hyodysenteriae PCR positive faecal samples helped confirm these as being from B. hyodysenteriae. This is the first time that B. hyodysenteriae has been detected in feral pigs. As these animals range over considerable distances, they present a potential source of B. hyodysenteriae for any domesticated pigs with which they may come into contact.     

Scanning electron microscopy and fluorescent in situ hybridization of experimental Brachyspira (Serpulina) pilosicoli infection in growing pigs

Two groups of six 8-week-old pigs were challenged with 1x10(9) cfu Brachyspira (Serpulina) pilosicoli or Serpulina intermedia daily for 3 consecutive days to study the pathology of porcine colonic spirochetosis by scanning electron microscopy (SEM) and fluorescent in situ hybridization (FISH) with oligonucleotide probes targeting ribosomal RNA specific for B. pilosicoli and the genus Brachyspira/Serpulina. Six pigs served as noninoculated controls. The animals were euthanatized successively between postinoculation days 14 and 24. B. pilosicoli was reisolated in feces from all of the inoculated pigs; however, only two pigs developed transient watery diarrhea. S. intermedia was reisolated from four of the inoculated pigs, but clinical signs were not observed. Gross examination of the B. pilosicoli-infected pigs revealed dilated large intestines with a hyperemic mucosa, whereas the large intestines of the S. intermedia-inoculated pigs and the control pigs appeared normal. SEM examination of B. pilosicoli-infected pigs revealed degenerated epithelial cells and spirochetal colonization of the colonic mucosa in four pigs. By FISH, B. pilosicoli cells were found colonizing and invading the surface epithelium and the crypts in all the pigs. Spirochetal crypt colonization markedly exceeded the occurrence of spirochetes on the mucosal surface. SEM examination of S. intermedia-inoculated pigs revealed no abnormalities, and Serpulina cells were detected only sporadically in the otherwise normal-appearing mucosa of four pigs by FISH. The results provide further evidence that B. pilosicoli is associated with colitis in pigs, although the gross lesions are mild. The spirochete is capable of colonizing the large intestine, inducing mucosal damage, invasion of the crypt and surface epithelium, and focal infiltration of the lamina propria. In addition, the study shows the applicability of FISH for specific identification of B. pilosicoli in formalin-fixed tissue.     

Analysis of bacterial load and prevalence of mixed infections with Lawsonia intracellularis, Brachyspira hyodysenteriae and/or Brachyspira pilosicoli in German pigs with diarrhoea

Lawsonia (L.) intracellularis, Brachyspira (B.) hyodysenteriae and B. pilosicoli are important pathogens in domestic pig production world-wide, responsible for porcine intestinal adenomatosis, swine dysentery, and porcine intestinal spirochetosis, respectively. Conventional PCR is the major diagnostic tool in the detection of the three pathogens, but the sole detection of bacterial DNA might lead to misinterpretations of results with respect to their clinical relevance, especially with mixed infections. Thus, the present study targeted the detection and quantification of the three pathogens in samples from herds with a case history of diarrhoea. Herds and samples were selected by the practitioners on a voluntary basis. Results were based on 1176 individual samples from 95 herds from Southern Germany. The pathogens were detected simultaneously by multiplex real-time PCR. The overall prevalence for L. intracellularis, B. hyodysenteriae and B. pilosicoli was 12.6%, 8.4% and 3.2% in faecal samples and 48.4%, 24.2% and 31.6% in herds, respectively. Sixty one percent, 82.6%, and 73.4% of herds positive for L. intracellularis, B. hyodysenteriae, and B. pilosicoli, respectively, had mixed infections. Median log values of DNA equivalents/g of faeces for L. intracellularis, B. hyodysenteriae and B. pilosicoli were 3.3, 5.9 and 3.2, with maxima of 8.3, 8.0 and 6.3, respectively. Within herd prevalence of B. hyodysenteriae and B. pilosicoli as well as the load of B. hyodysenteriae were significantly associated with the severity of diarrhoea.     

Influences of diet and vaccination on colonisation of pigs by the intestinal spirochaete Brachyspira (Serpulina) pilosicoli

The purpose of this study was to determine whether methods used to control swine dysentery (SD), caused by the intestinal spirochaete Brachyspira (Serpulina) hyodysenteriae, would also be effective in controlling porcine intestinal spirochaetosis (PIS) caused by the related spirochaete Brachyspira (Serpulina) pilosicoli. Weaner pigs in Groups I (n=8) and II (n=6) received a standard weaner pig diet based on wheat and lupins, whilst Group III (n=6) received an experimental diet based on cooked white rice and animal protein. Pigs in Group II were vaccinated intramuscularly twice at a 3-week-interval with a formalinised bacterin made from B. pilosicoli porcine strain 95/1000 resuspended in Freund's incomplete adjuvant. Eleven days later pigs in all groups were infected orally with 10(10) cells of strain 95/1000 on three successive days. One control pig in Group I developed acute diarrhoea, and at post-mortem had a severe erosive colitis with end-on attachment of spirochaetes to the colonic epithelium. All other pigs developed transient mild diarrhoea and had moderate patchy colitis at post-mortem 3 weeks later. B. pilosicoli was isolated from the faeces of all pigs, except for one fed rice, and was isolated from the mesenteric nodes of three pigs from Group I and from one vaccinated pig in Group II. Consumption of the rice-based diet, but not vaccination, delayed and significantly (p<0.001) reduced the onset of faecal excretion of B. pilosicoli after experimental challenge. Vaccination induced a primary and secondary serological response to B. pilosicoli, as measured using sonicated whole cells of strain 95/1000 as an ELISA plate coating antigen. Antibody titres in the vaccinated pigs then declined, despite intestinal colonisation by B. pilosicoli. Both groups of unvaccinated animals also failed to develop a post-infection increase in circulating antibody titres.     

Simultaneous detection of Brachyspira hyodysenteriae, Brachyspira pilosicoli and Lawsonia intracellularis in porcine faeces and tissue samples by multiplex-PCR

Diarrhoea in growing and finishing pigs is usually caused by infectious agents and laboratory diagnosis is a prerequisite for efficient therapy. Cultivation of Brachyspira hyodysenteriae or Brachyspira pilosicoli and detection of Lawsonia intracellularis by means of immunofluorescence tests (IFT) are time-consuming and in some cases lack sensitivity. A multiplex-PCR was designed to detect simultaneously these three pathogens in faeces and tissue samples, allowing the differential diagnosis of dysentery, intestinal spirochaetosis and proliferative enteropathy. Detection limits for B. hyodysenteriae, B. pilosicoli and L. intracellularis were 10(4), 10(2) and 10(3) copies respectively. Agreement between multiplex-PCR and nested-PCR or cultivation was considered substantial to almost perfect. Agreement between multiplex-PCR and IFT in detecting L. intracellularis was only moderate, which was probably related to false-positive results given by IFT. The multiplex-PCR described herein is a valuable tool for the rapid and simultaneous detection of three different pathogens in porcine samples causing enteric diseases.     

Occurrence of Lawsonia Intracellularis and Brachyspira spp. infection in swine suffering from diarrhoea

Principal aim of this study was to examine fecal samples from pigs suffering from diarrhea for the presence of Lawsonia intracellularis, Brachyspira hyodysenteriae and Brachyspira pilosicoli. The molecular techniques such as PCR and nested PCR were employed to detect the presence of p78 fragment of genomic DNA specific for Lawsonia intracellularis as well as fragment of tlyA gene specific for Brachyspira hyodysenteriae and 16S rDNA gene of Brachyspira pilosicoli. We assumed that about 25% of pigs were infected with Lawsonia intracellularis, about 10% with Brachyspira hyodysenteriae and only 0,8% with Brachyspira pilosicoli. In about 3% mixed infection with L. intracellularis and B. hyodysenteriae was observed. Results were comparable in herds that differed in quantity, breeding technology, hygienic standards and preventive treatment with different chemotherapeutics.     

Polyserositis in pigs caused by infection with Mycoplasma

A bovine adenovirus was isolated from a yearling heifer with systemic adenovirus infection. The virus was characterized as a subgroup 2 bovine adenovirus but could not be typed with reference antisera to the 10 known bovine adenovirus serotypes. A serological survey of cattle in the northern half of the North Island showed the prevalence of precipitating antibodies to bovine adenoviruses to be 43.3%.     

Epidemic of diarrhoea caused by porcine epidemic diarrhoea virus in Italy

There was an epidemic of diarrhoea affecting pigs of all ages in Italy between May 2005 and June 2006. In 63 herds the cause was confirmed as porcine epidemic diarrhoea virus by electron microscopy, immunoelectron microscopy, pcr and serology. Watery diarrhoea without mucus and blood was usually associated with a reduction of feed consumption. In farrowing-to-weaning herds, diarrhoea affected the sows and suckling piglets, and the mortality in newborn piglets was up to 34 per cent. In growers and fatteners the morbidity ranged from 20 to 80 per cent, but there was either no mortality or it was very low. Depending on the size of the herd and the type of operation, the clinical disease lasted for weeks or months.     

Pathologic changes caused by transplacental infection with an adenovirus-like agent in pigs

Five hysterectomy-derived colostrum-deprived pigs housed in individual cages with positive ventilation developed severe skin cyanosis and edema of the subcutaneous tissues in the submandibular, thoracic, and abdominal regions. The 5 pigs were killed 7 to 10 days after birth. Eosinophilic intranuclear inclusion bodies were found in endothelial cells of capillary and small blood vessels throughout the body. Ultrastructurally, nuclei of these affected endothelial cells contained small crystalline arrays of virus particles, which were considered to belong to the adenovirus-like group from their size and structure. The present results indicated that the porcine adenovirus-like agent might also have the ability to produce transplacental infection.     

The effects of experimental infection with bovine diarrhoea virus (BVDV) on lymphocyte subpopulations in the peripheral blood of pigs

The aim of the study was to analyse the dynamics of selected lymphocytes subpopulations in peripheral blood of pigs infected with BVDV, using flow cytometric method. The examinations were performed on eighteen healthy, pestivirus-free pigs divided into 3 groups. Pigs in the group 1 were intranasally infected with two virulent reference strains of BVDV: NADL2 and NADL8. Pigs of group 2 were included into experimentally infected ones. Group 3 consisted of control pigs free from infection. The percentage of CD2+ lymphocytes was gradually decreasing during the experiment. Finally a decrease by 20% was noted in group 2 and by 9% in group 1. A slight decrease of CD4+ cells and more significant decrease of CD8+ subpopulation were observed. The CD4:CD8 ratio changed from approximately 3:5 to 1:1 in pigs of group 2, and from 1:2 to 1:3 in pigs of group 1. There were only small fluctuations regarding TcR(gamma)delta+ cells. The level of these lymphocytes increased during first hours post infection and then decreased to the initial value. Obtained results may indicate that infection with species-unspecific pestiviruses causes similar, but less significant, changes within lymphocyte subpopulations than infection with species-specific pestiviruses.