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The intention to increase roach (Rutilus rutilus) consumption is in focus for ecological and economic reasons in Finland. However, its safety as food has not been considered comprehensively. We collected and artificially digested 85 roach halves originating from the south-eastern coast of Finland, and found trematode metacercariae in 98.8% of the samples. Based on polymerase chain reaction (PCR) and sequencing of amplicons generated from the ITS2 gene region, zoonotic parasites of the family Opistorchiidae were identified as Pseudamphistomum truncatum and Metorchis bilis, and also non-zoonotic Holostephanus dubinini (family Cyathocotylidae) and Posthodiplostomum spp. (family Diplostomidae) were identified. The species identity of other trematodes found is currently being investigated. Mixed infections of several trematode species were common. The prevalence of morphologically identified zoonotic P. truncatum was 46%, and zoonotic M. bilis was found in one sequence sample. The high prevalence of zoonotic trematode metacercariae in roach from the Gulf of Finland is alarming. Only thoroughly cooked roach products can be recommended for human or animal consumption from the area.

The present study reports the effect of Penicillin G (PenG) on the proteome dynamics of the Staphylococcus aureus strain Newman during biofilm mode of growth. The viability of the 18-h-old biofilm cells challenged with PenG at the concentration of 1mgmL(-1) was first assessed by plate counting, resazurin and LIVE/DEAD fluorescence staining, which indicated that the viability was reduced by ∼35% and ∼90% at 2h and 24h, respectively, after the addition of PenG. Subsequent two-dimensional difference gel electrophoresis (2D DIGE) assay of the treated and non-treated biofilm cells at the indicated time points revealed 45 proteins showing time- and treatment-specific change (1.5-fold, p

English summary

One-hundred-and-thirty-nine fecal samples were examined to assess the prevalence of Parascaris spp. and strongyle infections in two-year-old or younger horses in Finland. The owners of the horses were asked to answer an online questionnaire about the horses' environment and the management practices of the stable. The results of fecal examination and the survey were analyzed to evaluate the effect of different risk factors as ascertained by the survey on parasite prevalence. The prevalence of Parascaris spp. infections at 11.5% was lower than expected based on previous research and the strongyle prevalence of 57.6% was found in young Finnish horses. Strongyloides westeri and Eimeria leuckarti infections were also found. Pasture hygiene had a stronger influence on the prevalence of strongyle infections than on Parascaris spp. infections, whereas the hygiene routine of the horses' housing was found to be more important in the prevention of Parascaris spp. infections. The planning of the control of parasitic infections should be based on the identified risk factors.

Staphylococcus epidermidis (SE) includes commensal and pathogenic strains capable of infecting humans and animals. This study reports global exoproteome profiling of bovine mastitis strain PM221 and two human strains, commensal-type ATCC12228 and sepsis-associated RP62A. We identified 451, 395, and 518 proteins from culture supernatants of PM221, ATCC12228, and RP62A, respectively. Comparison of the identified exoproteomes revealed several strain-specific differences related to secreted antigens and adhesins, higher virulence capability for RP62A, and similarities between the PM221 and RP62A exoproteomes. The majority of the identified proteins (∼80%) were predicted to be cytoplasmic, including proteins known to be associated in membrane vesicles (MVs) in Staphylococcus aureus and immunogenic/adhesive moonlighting proteins. Enrichment of MV fractions from culture supernatants and analysis of their protein composition indicated that this nonclassical protein secretion pathway was being exploited under the conditions used and that there are strain-specific differences in nonclassical protein export. In addition, several predicted cell-surface proteins were identified in the culture media. In summary, the present study is the first in-depth exoproteome analysis of SE highlighting strain-specific factors able to contribute to virulence and adaptation.

The study aimed to determine the effects of red clover (Trifolium pratense) silage with high phyto-oestrogen content on ewe performance during their first breeding season. Red clover silage containing formononetin, biochanin A, genistein, and daidzein was fed to 10 nulliparous ewes of the prolific Finnish Landrace breed before, during and after the breeding season, for a total of 5 months. A control group of 10 ewes was fed with grass silage. The mean numbers of foetuses per pregnancy were 2.1±0.7 and 2.2±0.8 for the red clover and control groups, respectively. The total mass of the uterus with its contents was significantly greater in ewes of the red clover group compared with those of the control group. This difference was mainly explained by the greater volume of foetal fluids. Serum progesterone concentration in the red clover group was significantly lower over the entire period analysed than in the control group. In conclusion, the fecundity of the ewes was not reduced by red clover feed with high phyto-oestrogen concentrations. The volume of foetal fluids increased that could increase the risk for vaginal prolapse before the term.

Trichinella spiralis causes a significantly higher parasite burden in rat muscle than Trichinella nativa. To assess whether the difference in infectivity is due to the early intestinal response, we analyzed gene expression changes in the rat jejunum during Trichinella infection with a whole-genome microarray. The rats were euthanized on day five of infection, and their jejunal mucosa was sampled for microarray analysis. In addition, intestinal histology and hematology were examined. Against our expectations, the gene expression changes were similar in both T.nativa- and T. spiralis-infected groups. The two groups were hence pooled, and in the combined Trichinella-infected group, 551 genes were overexpressed and 427 underexpressed when compared to controls (false discovery rate ≤ 0.001 and fold change at least 2 in either direction). Pathway analysis identified seven pathways significantly associated with Trichinella infection (p0.05). The microarray data suggested nonspecific damage and an inflammatory response in the jejunal mucosa. Histological findings, including hyperemia, hemorrhage and a marked infiltration of inflammatory cells, supported the microarray data. Trichinella infection caused complex gene expression changes that indicate a host response to tissue damage in the mucosa of the jejunum, but the changes were not notably dependent on the studied species of Trichinella.

We investigated the presence of Toxoplasma gondii infections in Eurasian lynx (Lynx lynx) in Finland by analyzing samples from 337 lynx that were legally hunted during the 2010-2011 season and by performing a retrospective nationwide database search of postmortem toxoplasmosis diagnoses in this species. We detected specific anti-T. gondii IgG antibodies in 290 (86.1%) of the 337 lynx. The method used was a direct agglutination test, and samples positive at the used dilution 1:40 were defined as antibody positive. Older lynx had 14.3 times higher odds of being antibody-positive than did lynx of the presumed age of 7-10 mo, and lynx weighing15 kg had 16.7 times higher odds of being antibody positive than did those ≤ 15 kg. Lynx from the southwest were more often antibody positive, with an odds ratio 6.3, than lynx from the northeast. None of the 332 fecal samples available was positive for the presence of T. gondii-like oocysts with a quantitative MgSO4 flotation technique, and none of the 167 free-ranging Eurasian lynx examined postmortem by veterinary pathologists from January 2000 to May 2010 had died from toxoplasmosis. Although Finnish lynx were confirmed to commonly encounter T. gondii, we found no evidence of an ongoing contribution to the environmental oocyst burden nor of the lynx dying from the infection.

Chronic osteoarthritis (OA) is a degenerative disease of the articular cartilage. DNA-binding high mobility group protein B1 (HMGB1) is released on cellular death/activation and acts as an endogenous danger signal and a proinflammatory cytokine. Matrix metalloproteinase (MMP)-2 and in MMP-9 are induced to mediate proteolytic degradation/remodelling of joint tissues. Collagen degradation in the bone and synovium leads to release of type I collagen-derived cross-linked carboxy-terminal telopeptide (ICTP). These molecules have been linked to the pathogenesis of OA and could have potential as synovial fluid (SF) biomarkers in OA. Cartilage and SF were obtained from 27 dairy bulls (30-61 months old) and control cartilage from six young healthy dairy bulls. OA lesions were evaluated grossly (five grades), histologically (seven Osteoarthritis Research Society International [ORSI] grades) and immunohistochemically (four HMGB1 grades). The OARSI lesion score was calculated as the product of the OARSI grade and the OARSI score (the total area of the lesions). SF concentrations of HMGB1, MMP-2 and -9 and ICTP were measured by enzyme-linked immunosorbent assay, gelatin zymography and radioimmunoassay, respectively. Seventy-two percent (39/54) of stifle joints and 85% (23/27) of the dairy bulls had at least one gross OA lesion and 94% of the lesions were localized to the distal end of the femur, with the patellar groove and the lateral trochlear ridge being predilection sites. Gross and histological grades correlated with the HMGB1 grade, but SF total cell count, percent neutrophils or the measured biomarkers did not correlate with the tissue lesions, with the exception of ICTP concentration, which correlated with the total joint score. The switch of HMGB1 from DNA-binding nuclear protein to an extracellular alarmin/cytokine correlates with the gross and histological grades of OA tissue lesions. However, the activity and extent of the tissue lesions did not correlate with other SF biomarkers, perhaps because the histological grades represent outcome measures, while SF reflects process parameters. The only exception was ICTP concentration, which reflects enhanced destruction/remodelling.

We sampled 339 fecal samples, 296 intestines, and 82 lungs from 371 lynx hunted during the 2010-2011 season in Finland. The fecal samples were analyzed for endoparasites by a quantitative flotation method, and helminths from intestines were studied morphologically, while lungs were investigated for pulmonary parasites. From fecal samples, eggs and oocysts of at least 6 different endoparasite species were identified, with a mean of 1.5 (range 0-4) parasite species per host. In the intestines, at least 4 different helminth species were found, with the mean of 2.0 (range 1-4) species per infected host. The prevalence of eggs in feces and the prevalence of worms in intestines were 71% and 93% for Toxocara cati , 29% and 68% for Taenia spp., and 5% and 2% for Diphyllobothrium sp., respectively. Only eggs were detected for Capillaria sp. (46%) and Uncinaria sp. (0.6%) nematodes, and only adults were detected for Mesocestoides sp. cestodes (0.3%). Significant positive correlations were evident between the number of T. cati (r = 0.664; P = 0.01) and Diphyllobothrium sp. (r = 0.645; P = 0.01) eggs per gram of feces and adult worms detected in intestine. In addition to the metazoan parasites, protozoan Isospora sp. oocysts were also found (0.6%). Pulmonary samples were all negative for parasites. These data demonstrate that lynx commonly harbor various endoparasites, some of which are zoonotic.

SUMMARYCestodes of the genus Taenia are parasites of mammals, with mainly carnivores as definitive and herbivores as intermediate hosts. Various medium-sized cats, Lynx spp., are involved in the life cycles of several species of Taenia. The aim of the present study was to identify Taenia tapeworms in the Eurasian lynx (Lynx lynx) from Finland. In total, 135 tapeworms from 72 lynx were subjected to molecular identification based on sequences of 2 mtDNA regions, the cytochrome c oxidase subunit 1 and the NADH dehydrogenase subunit 1 genes. Available morphological characters of the rostellar hooks and strobila were compared. Two species of Taenia were found: T. laticollis (127 samples) and an unknown Taenia sp. (5 samples). The latter could not be identified to species based on mtDNA, and the rostellar hooks were short relative to those described among other Taenia spp. recorded in felids from the Holarctic region. In the phylogenetic analyses of mtDNA sequences, T. laticollis was placed as a sister species of T. macrocystis, and the unknown Taenia sp. was closely related to T. hydatigena and T. regis. Our analyses suggest that these distinct taeniid tapeworms represent a putative new species of Taenia. The only currently recognized definitive host is L. lynx and the intermediate host is unknown.

The detection in the early 2000's of new, pandemically spreading viral diseases and threats led to "One Health", a holistic concept of the inevitability of collaboration between human and animal health and the protection of the ecosystem. The movement initiated by physicians and veterinarians emerges form the idea that the health of humans and animals is interconnected and connected with the environment and that changes occurring in the environment will have a significant impact on health. Problems associated with health, such as antimicrobial resistance or zoonoses, require global solutions.

The meat of wild boar (Sus scrofa L.) can be a source of human infections with zoonotic parasites Toxoplasma gondii and Trichinella spp. We screened 197 wild boar sera collected at slaughter from 25 Finnish farms in 2007-2008 for serological evidence of infections with these parasites. Using a commercial direct agglutination test at a serum dilution of 1:40, T. gondii-specific IgG antibodies were detected in 65 (33.0%) samples, on 14 (56.0%) farms. Females, animals older than 24 months, animals of small herds, and animals originating from south-western parts of Finland were more often T. gondii-seropositive than were males, younger animals, animals of larger herds, and animals originating from the north and east, respectively. Four (2.0%) of the sera, originating from three (12.0%) farms, tested Trichinella-seropositive with an in-house ELISA and a conservative cut-off for seropositivity. One farm had both T. gondii- and Trichinella-seropositive animals. Taken together, an infection source had been present on 16 (64.0%) farms, and 69 (35.0%) of the 197 farmed wild boars intended for human consumption had specific serological evidence of exposure to a zoonotic parasite.

The aim of this study was to estimate the prevalence of feline intestinal parasites in Finland and to determine the possible risk factors for infection. Altogether 411 feline fecal samples were analyzed with a flotation method to reveal helminth eggs and protozoan oocysts. Of the samples, 402 were also screened for Giardia species antigens with a commercial enzyme-linked immunosorbent assay kit. The cat owners completed a questionnaire. Toxocara cati prevalence was 5.4% and Toxascaris leonina 0.2%. Taenia species eggs were found in 1.5% of the samples and Isospora felis in 0.7%, whilst 3.2% of the samples tested positive for Giardia species antigen. Risk factors for Toxocara/ Toxascaris species infection included being a non-pedigree cat, having access to the outdoors, living outside of the cities and receiving home-made food. Pedigree cats were at greater risk of contracting Giardia duodenalis. The majority of the cat owners (62.4%) treated their cat with anthelmintics 2–4 times per year.

In Staphylococcus aureus, ClpP proteases were previously shown to be essential for virulence and stress tolerance in strains derived from NCTC8325. Because these strains exhibit a severely reduced activity of the alternative sigma factor, SigB, we here reassessed the role of ClpP in SigB-proficient clinical strains. To this end, clpP was deleted in strains COL, Newman, and SA564, and the strains were characterized phenotypically. The proteomic changes accomplished by the clpP deletion in the different strains were analyzed using the 2-D DIGE technique. The proteomic analyses revealed mostly conserved changes in the protein profiles of the ClpP-deficient strains. Among the strain-specific changes were the up-regulation of prophage proteins that coincided with an increased spontaneous release of prophages and the relatively poorer growth of the clpP mutants in some strain backgrounds. Interestingly, the effect of ClpP on the expression of selected virulence genes was strain-dependent despite the fact that the expression of the global virulence regulators RNAIII, mgrA, sarZ, sarR, and arlRS was similarly changed in all clpP mutants. ClpP affected the expression of sarS in a strain-dependent manner, and we propose that the differential expression of sarS is central to the strain-dependent effect of ClpP on the expression of virulence genes.

Streptococcus uberis is an environmental udder pathogen that infects cattle and can cause persistent intramammary infection (IMI), despite the fact that isolates are mainly susceptible to antibiotics. As biofilm growth can cause persistent infection, the ability of ten S. uberis isolates from clinical and subclinical IMIs to form biofilms on the polystyrene surface of a conventional 96-microplates model was examined. Biofilm formation was judged by different staining methods (crystal violet and resazurin) and by atomic force and fluorescence microscopy. These analyses revealed that two out of ten S. uberis strains tested were able to form biofilms. Upon treatment with Proteinase K, biofilms of S. uberis were completely disintegrated, which indicates that biofilm formation is protein-mediated in these strains. Addition of trace amounts of milk, the natural growth medium of S. uberis, significantly increased biofilm formation by most of the strains initially classified as non-biofilm producers. Alpha-casein and β-casein were the primary inducers of biofilm growth, and casein degradation by serine protease activity was required to achieve maximal biofilm production. These results suggest that the extracellular proteolytic activity of S. uberis contributes to an increased biofilm formation. Such a mode of growth induced by host proteins might help to explain the persistence of IMIs caused by this pathogen.

Although human infections caused by Trichinella sp. have not been reported in Finland for several decades and Trichinella sp. infection in pork has become virtually extinct in the last decade, sylvatic Trichinella spp. infection is still highly prevalent in Finland. Muscle digestion of 2,483 carnivorous wild animals from 9 host species during 1999-2005 showed 617 positive animals (24.8%). Molecular identification from 328 larval isolates revealed 4 different endemic Trichinella species, i.e., T. nativa, T. spiralis, T. britovi, and T. pseudospiralis. Seven percent of the infected animals carried mixed infections. Trichinella nativa was the most common species (74%), but T. spiralis was identified in 12%, T. britovi in 6%, and T. pseudospiralis in 1% of the animals. Host species showed different sample prevalence and Trichinella species distribution. Geographical distribution also varied, with the southern part of the country having significantly higher percentages than the northern part. Infection density was dependent on both the infecting Trichinella species and the host species. Trichinella spiralis was discovered in areas with no known domestic infection cases, indicating that it can also occur in the sylvatic cycle. Raccoon dogs and red foxes are the most important reservoir animals for T. spiralis , as well as for the sylvatic Trichinella species in Finland.

A central primitive neuroectodermal tumour (PNET) with ependymal differentiation was identified in an 18-month-old female Australian terrier dog that died following signs of central nervous system disease. A large space-occupying haemorrhagic mass replaced the thalamus and part of the hypothalamus of the brain. Microscopical examination revealed a tumour formed by a compact sheet of small cells that contained ependymal and perivascular pseudorosettes. Immunohistochemically, the neoplastic cells variably expressed glial fibrillary acidic protein, neuron-specific enolase, vimentin and cytokeratin, consistent with divergent differentiation of the tumour. This case is thought to represent the first description of PNET with ependymal differentiation in a dog.

A 2-month-old mongrel dog had multifocal necrotizing hepatitis, interstitial pneumonia, and hemorrhagic enteritis. Immunohistochemistry detected antigens of Clostridium piliforme in the intestine and liver, and antigens of canine distemper virus within the lung, urinary bladder, brain, spleen, and liver. Furthermore, uncharacterized intralesional coccidian protozoa were observed within the intestine.

In the period 1996-2006 two specific sperm defects, the knobbed acrosome (KA) defect and the immotile short-tail sperm (ISTS) defect, showed a strong negative association with fertility in Finnish breeding boars. In this study, we examined the incidence of these two sperm defects in two pig breeds, their effects on fertility and their associations with sperm morphology and testicular histology. Semen samples from 2048 (1097 Yorkshire, 951 Landrace) boars were collected. None of the Landrace boars revealed either the KA defect or the ISTS defect. Of the Yorkshire boars, 0.8% were afflicted with the KA defect and 7.6% with the ISTS defect. Boars diagnosed with the ISTS defect produced no litters. Fertility data were available from two artificially inseminated (AI) boars and six farm breeding boars affected with the KA defect. Breeding boars with 45-81% knobbed spermatozoa (n = 6) did not produce any litters out of 71 sows bred. AI boars with 25-30% knobbed spermatozoa had a poor non-return rate (on average 47% compared with 85% for normal control boars) and produced small litters, on average 2.5 piglets less than other boars of the same breed. Morphometry of testicular tissue and distribution of different cells in the seminiferous tubules were examined in nine boars. Boars with the KA defect had a smaller diameter of the seminiferous tubules (p < 0.05) and a lower number of Sertoli cells (p < 0.05) than controls. ISTS boars, in turn, had a significantly lower number of elongated spermatids (p < 0.05), and they also produced on average only 12% of the spermatozoa of normal boars. The ISTS defect is a manifestation of an autosomal recessive disease caused by an insertion in the KPL2 gene in porcine chromosome 16. Although we tried to map the KA defect, its aetiology remains unclear.

Trichinella spp. can infect various domestic and wild species, including companion animals. Infection occurs because of the ingestion of raw meat (e.g., infected prey). In experimental studies, cats have been found to be a very susceptible host to infection by Trichinella spp.; naturally occurring feline infections have also been reported. However, clinically apparent disease seems to be a rare manifestation of this infection in cats. The skin biopsy of an 8-year-old, neutered, male, domestic cat revealed an inflammatory granulation tissue that surrounded a well-preserved cyst that contained a Trichinella sp. larva. Distinct seropositive reaction against Trichinella spp. antigens was demonstrated by enzyme-linked immunosorbent assay and Western blot. Immunohistochemistry, by using serum from the infected cat as the source of antibody, showed strong immunostaining of Trichinella spp. larvae. During a 1-year follow-up, a postexcisional local tissue reaction was observed. This manifested as a firm, poorly circumscribed subcutaneous mass adjacent to the eye, which demonstrated clinical features and histopathologic findings indicative of chronic inflammation associated with granulation tissue and fibrodysplasia. Digestion of the muscle biopsy revealed one Trichinella sp. larva, which was identified by multiplex polymerase chain reaction as Trichinella nativa. To the authors' knowledge, this is the first documented case of trichinellosis in a cat with a nonhealing ulcerative skin lesion as the main clinical manifestation of the infection.

Harmful parasites of the wild northern boreal mammals are still surprisingly poorly studied. In 2003-2006, a peritonitis outbreak caused by the filarioid nematode, Setaria tundra, emerged in Finland's reindeer population. In order to gain knowledge about the basic biology, epidemiology, and transmission dynamics of this parasite, samples for S. tundra were collected from reindeer and other cervids during the follow-up period 2004-2006. Using morphology and molecular biology methods, we describe here S. tundra's first larval stage, microfilaria (smf), for the first time scientifically. The prevalence and densities of smf were higher in reindeer calves than in adults, overall prevalence being 42%. The overall smf prevalences for moose, wild forest reindeer and roe deer were 1.4-1.8%, 23%, and 39%, respectively. The focus of microfilaremia moved north and settled down in the south simultaneously with the peritonitis outbreak. The peak microfilaremia occurred in the first summer after the infection, and smf disappeared from the blood after 2 years. Captive reindeer were smf positive over the year. The prepatent period of S. tundra was estimated to be about 4 months, and the life span at least 14 months. This parasite likely has an important impact on boreal ecosystems.

Streptococcus uberis is an environmental bovine mastitis pathogen capable of UV-inducible SOS mutagenesis. Bacterial SOS systems can be induced by several chemicals including also antibiotics used in clinical practice. Here, we have studied the effect of ciprofloxacin, a fluoroquinolone antibiotic and known inducer of SOS, on mutations leading to antibiotic resistance in S. uberis. Mutation frequencies and spectra were compared in a wild-type S. uberis strain and its DeltaumuC derivative. The results revealed that concentrations of ciprofloxacin corresponding to 0.3-0.5x minimum inhibitory concentration (MIC) induce mutagenesis independent of UmuC. Partial sequencing of the rpoB gene of individual rifampin-resistant clones from wild-type and DeltaumuC strains revealed a similar but complex pattern of point mutations including transitions, transversions and deletions/insertions. It was previously shown that UV induces mainly transition-type mutations and UmuC is essential for the process. Thus, the results presented here demonstrate that S. uberis employs distinct mechanisms for ciprofloxacin and UV-induced mutagenesis, which is a striking difference to Escherichia coli SOS model.

SUMMARYThe first reported bovine botulism outbreak in Finland is described. Nine out of 90 cattle on a dairy farm died after being fed non-acidified silage contaminated by animal carcasses. Type C botulinum neurotoxin gene was detected in one heifer by polymerase chain reaction (PCR) and the neurotoxin was detected by the mouse bioassay.Clostridium botulinumtype C was isolated from liver samples. The isolated strain was identified with amplified fragment length polymorphism (AFLP) analysis as group IIIC. botulinum. To our knowledge, this is the first time that a type C bovine botulism outbreak has been diagnosed by PCR and confirmed by subsequent isolation and AFLP identification of the disease strain. The importance of the acidification process in silage production to inhibitC. botulinumtoxin production in silage and thus to prevent further botulism outbreaks is emphasized. Nevertheless, preformed toxin in the carcass is not destroyed by acid.

OBJECTIVE: Human immunodeficiency virus (HIV)-seropositive patients have frequently severe gingival inflammation and/or attachment loss. In addition many infectious diseases affect their periodontium with varying clinical manifestations. Matrix metalloproteinases seem to play a key role in physiological periodontal remodelling and pathological tissue destruction. The aim of the present study was to characterize the presence, molecular forms, cellular sources, activities, and relative amounts of fibroblast-type (matrix metalloproteinase [MMP]-1) and neutrophil (MMP-8) collagenases, as well as their potential activator stromelysin-I (MMP-3) and myeloperoxidase in saliva of HIV-seropositive patients at different phases of HIV-infection. HIV-seronegative, healthy, age-matched patients served as controls. PATIENTS AND METHODS: Saliva samples were characterized by Western blotting using antibodies specific for MMP-1, MMP-3 and MMP-8. Interstitial collagenase activities were measured using quantitative sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis/laser densitometry assay. Myeloperoxidase was analysed using quantitative dot blotting. RESULTS: Clinical and microbiological evaluation of HIV-seropositive patients' periodontium showed the presence of putative periodontopathogens ie Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), Prevotella intermedia (Pi), Peptostreptococcus micros (Psm) and Campylobacter rectus (Cr) in their periodontal pockets. The amount of Candida increased with the severity of HIV-infection. Clinical and microbiological findings of HIV-seropositive patients suggested that they have a tendency to develop periodontal disease. Interstitial collagenase activities were found to be increased in saliva of different phases of HIV-infected patients compared to the controls. Independent of the phase of HIV-infection saliva samples contained pro- and active forms of MMP-1, -3 and -8 using Western blotting. Saliva samples from healthy controls were found to contain hardly any immunoreactivities for MMP-1 or MMP-8, but considerable amounts of MMP-3 were detected. Quantitative dot blotting demonstrated increased amounts of myeloperoxidase in HIV-patients' saliva relative to controls. CONCLUSION: The present results showed increased amounts of MMP-1, -3, -8 and myeloperoxidase in HIV-patients' saliva. MMP-1 and -8 may have been activated by MMP-3 and/or oxidants generated by myeloperoxidase. The increased amounts of MMPs and myeloperoxidase may reflect and directly participate in HIV-infection associated periodontitis.

Study design Proteinases were immunohistochemically stained to analyze degenerated discs and paradiscal tissues in comparison to contiguous control tissues in an experimental porcine model of intervertebral disc degeneration. Objective The aim was to analyze plasmin and metalloproteinases known to participate in mutual activation cascades. Summary of background data Comparison of the degenerated discs and paradiscal structures with control tissues disclosed accumulation of plasmin and induction of matrix metalloproteinases (MMP), MMP-1 and MMP-2 in the discs, but some other MMPs in reactive and remodeling tissues. Material and methods In 6 domestic pigs, the cranial L4 endplate was perforated to penetrate the nucleus pulposus. Three months later, the animals were killed and the experimental and the contiguous control vertebrae, complete with their intervertebral discs, were excised and subjected to histologic and immunohistochemical examinations. Results Immunohistochemical analysis disclosed increased expression of MMP-1 and MMP-2 in the traumatized and degenerated intervertebral discs. Some MMPs were also induced in all paradiscal structures (bone marrow, vertebral bone, and spinal ligaments), or decreased in already scarred areas. The common denominator for all the anatomic sites studied was accumulation of plasmin. Conclusion Fibroblast collagenase (MMP-1) and gelatinase A (MMP-2), capable of degrading native and denatured collagen, were induced in degenerating intervertebral discs. Use of an experimental model enabled demonstration that biomechanical destabilization and degeneration of the disc also affects all other paradiscal structures, which are subjected to proteolysis and/or reparative fibrosis apparently representing remodeling of the spine subjected to pathologic stress. Profiling of various MMPs and plasmin, known to participate in mutual activation cascades, suggests that plasmin could activate pro-MMP-1, pro-MMP-2, pro-MMP-3, pro-MMP-7, pro-MMP-9, and pro-MMP-13 and alone or/and in cooperation with MMP-3 initiate at least 2 mutual MMPs activation cascades driven by activated MMP-3 and MMP-7.

Fumonisin B(1) (FB(1)), the most prevalent member of toxins produced by several species of Fusarium molds, which occur mainly in maize, causes several fatal hepatopathies and nephropathies of animals. The current study was scrutinized to ascertain different cytotoxic and morphological transformations in rat hepatocytes induced by the treatments of diverse concentrations (300, 500, or 1000 microM) of fumonisin B(1) in vitro, using both monolayer and spheroid cultures. In each hepatocyte culture, the cytotoxicity of FB(1) was augmented in dose- and time-response manners. Morphological transformations among FB(1)-treated groups integrated accumulation of lipid droplets, cytoplasmic vacuolation in hepatocyte monolayers, and bleb formation in the hepatocyte spheroids. Additionally, electron microscopy revealed the loss of microvilli, mitochondrial swelling, and formation of lamellar membranous whorl in the vacuoles and bile canaliculi-like structures. Appearance of electron dense bodies in the monolayers, and loss of cell-to-cell contact in spheroids were depicted in 1000 microM FB(1)-treated hepatocytes. These outcomes insinuate different vital events in explaining morphological transformations in the cell membrane and organelles, induced by fumonisins in rat hepatocytes.

This article describes the ECVP/ESVP Summer School in Veterinary Pathology, a new annual two-week European training facility established by the European College of Veterinary Pathologists (ECVP) in collaboration with the European Society of Veterinary Pathology (ESVP). The aim of the Summer Schools is to provide Europe-wide, harmonized, top-standard theoretical and practical post-graduate training for veterinarians specializing in veterinary pathology. In particular, it aims to support trainees in veterinary pathology in their individual preparation for the ECVP certifying examination. Ultimately, it aims to provide young pathologists with the skills and knowledge necessary to participate in international, high-quality research and the tools for applying international standards to their own research and for independent study for the ECVP certifying examination, even if they do not work in comparable academic environments and do not have the same level of local support and training. The ECVP/ESVP Summer Schools take place in European countries, with local organization from a university department of veterinary pathology. Each event comprises modules provided by internationally recognized specialists in their specific fields of expertise on different organ systems, diseases of specific species, specific techniques, and specific topics relevant to pathology, forming a cycle of four events to cover all major topics. Every two years a mock exam is organized as a tool to monitor individual progress in preparing for the ECVP certifying examination.