Laboratorio ja utaretulehdus

Summary: Laboratory and mastitis

Human-to-bovine jump of Staphylococcus aureus CC8 is associated with the loss of a β-hemolysin converting prophage and the acquisition of a new staphylococcal cassette chromosome.

Staphylococcus aureus can colonize and infect both humans and animals, but isolates from both hosts tend to belong to different lineages. Our recent finding of bovine-adapted S. aureus showing close genetic relationship to the human S. aureus clonal complex 8 (CC8) allowed us to examine the genetic basis of host adaptation in this particular CC. Using total chromosome microarrays, we compared the genetic makeup of 14 CC8 isolates obtained from cows suffering subclinical mastitis, with nine CC8 isolates from colonized or infected human patients, and nine S. aureus isolates belonging to typical bovine CCs. CC8 isolates were found to segregate in a unique group, different from the typical bovine CCs. Within this CC8 group, human and bovine isolates further segregated into three subgroups, among which two contained a mix of human and bovine isolates, and one contained only bovine isolates. This distribution into specific clusters and subclusters reflected major differences in the S. aureus content of mobile genetic elements (MGEs). Indeed, while the mixed human-bovine clusters carried commonly human-associated β-hemolysin converting prophages, the bovine-only isolates were devoid of such prophages but harbored an additional new non-mec staphylococcal cassette chromosome (SCC) unique to bovine CC8 isolates. This composite cassette carried a gene coding for a new LPXTG-surface protein sharing homologies with a protein found in the environmental bacterium Geobacillus thermoglucosidans. Thus, in contrast to human CC8 isolates, the bovine-only CC8 group was associated with the combined loss of β-hemolysin converting prophages and gain of a new SCC probably acquired in the animal environment. Remaining questions are whether the new LPXTG-protein plays a role in bovine colonization or infection, and whether the new SCC could further acquire antibiotic-resistance genes and carry them back to human.

Utaretulehduksen hoito lypsykaudella

Summary: Treatment of mastitis during lactation

Umpeenpanohoito utaretulehdusten hoidossa ja ennaltaehkäisyssä

Dry cow therapy in the treatment of mastitis and prevention of new intramammary infections

Koagulaasinegatiivisten stafylokokkien aiheuttama utaretulehdus lehmällä

Summary: Mastitis caused by CNS in the cow

Plant compounds insecticide activity against Coleoptera pests of stored products

The objective of this work was to screen plants with insecticide activity, in order to isolate, identify and assess the bioactivity of insecticide compounds present in these plants, against Coleoptera pests of stored products: Oryzaephilus surinamensis L. (Silvanidae), Rhyzopertha dominica F. (Bostrichidae) and Sitophilus zeamais Mots. (Curculionidae). The plant species used were: basil (Ocimum selloi Benth.), rue (Ruta graveolens L.), lion's ear (Leonotis nepetifolia (L.) R.Br.), jimson weed (Datura stramonium L.), baleeira herb (Cordia verbenacea L.), mint (Mentha piperita L.), wild balsam apple (Mormodica charantia L.), and billy goat weed or mentrasto (Ageratum conyzoides L.). The insecticide activity of hexane and ethanol extracts from those plants on R. dominica was evaluated. Among them, only hexane extract of A. conyzoides showed insecticide activity; the hexane extract of this species was successively fractionated by silica gel column chromatography, for isolation and purification of the active compounds. Compounds 5,6,7,8,3',4',5'-heptamethoxyflavone; 5,6,7,8,3'-pentamethoxy-4',5'-methilenedioxyflavone and coumarin were identified. However, only coumarin showed insecticide activity against three insect pests (LD50 from 2.72 to 39.71 mg g-1 a.i.). The increasing order of insects susceptibility to coumarin was R. dominica, S. zeamais and O. surinamensis.

Tumor necrosis factor-alpha mediates changes in tissue protein turnover in a rat cancer cachexia model.

Rats bearing the Yoshida AH-130 ascites hepatoma showed enhanced fractional rates of protein degradation in gastrocnemius muscle, heart, and liver, while fractional synthesis rates were similar to those in non-tumor bearing rats. This hypercatabolic pattern was associated with marked perturbations of the hormonal homeostasis and presence of tumor necrosis factor in the circulation. The daily administration of a goat anti-murine TNF IgG to tumor-bearing rats decreased protein degradation rates in skeletal muscle, heart, and liver as compared with tumor-bearing rats receiving a nonimmune goat IgG. The anti-TNF treatment was also effective in attenuating early perturbations in insulin and corticosterone homeostasis. Although these results suggest that tumor necrosis factor plays a significant role in mediating the changes in protein turnover and hormone levels elicited by tumor growth, the inability of such treatment to prevent a reduction in body weight implies that other mediators or tumor-related events were also involved.

Klebsiella utaretulehduksen aiheuttajana : kirjallisuuskatsaus ja tapausselostus

Summary: Klebsiella as mastitis causing agent : literature review and a case report

Tumor localization of radiolabeled antibodies against carcinoembryonic antigen in patients with carcinoma: a critical evaluation.

Purified, [131I]-labeled goat antibodies against carcinoembryonic antigen, which have been shown to localize in human carcinoma in nude mice, were injected into 27 patients with carcinoma. Patients were scanned with a scintillation camera at various intervals. In 11 patients, radioactivity was detectable in the tumor 48 hours after injection. Computerized subtraction of blood-pool radioactivity provided clearer pictures in positive cases, but in 16 patients the scans remained doubtful or negative. To study the specificity of [131I]-antibody localization, we gave some patients simultaneous injections of [125I]-labeled normal IgG. Both isotopes were measured by means of scintillation counting in tumors and normal tissues recovered after surgery. The results demonstrated that only the anti-CEA antibodies localized in tumors. However, the total antibody-derived radioactivity in the tumor was only about 0.001 of the injected dose. We conclude that, despite the present demonstration of specificity, this method of tumor detection is not yet clinically useful.

Transmission of MRSA and MSSA between humans and farm animals

Human nares are the main niche of Staphylococcus aureus, but farm animals can be also infected (cows) or colonized (pigs) constituting significant reservoir of this pathogen. Previous studies indicated that human and animal strains are quite distinct but the extent of cross-species specialization and transmission remains largely unknown. However, recent reports from several European countries as well as USA and Canada have indicated that employment in farming is an emerging risk factor for MRSA carriage. Pigs were found to be frequently colonized with MRSA, usually with a strain belonging to CC398. It is not known whether animal-human transmission was specific to this particular MRSA strain. S. aureus isolates from cow mastitis and pig colonization isolates were collected in parallel to nasal swab isolates from the animals' caretakers. The isolates were genotyped by AFLP, spatyping, and when appropriate by MLST. The isolates from cow mastitis were genetically uniform in comparison with human isolates. They were quite distinct from farmers\' carriage isolates, indicating pronounced hostspecialization. However, several cases where an infected cow and a colonized farmer had the same strain were detected, including one farm where two farmers were colonized and two cows were infected with MRSA belonging to CC398. Pig isolates were genetically more diverse than cow isolates. They were different from both human and cow isolates with one notable exception. Large fraction of pigs (20%) and pig caretakers (50%) were colonized with isolates belonging to CC398, majority of which were MSSA (2 cases of MRSA). These results indicate that host specialization in S. aureus is quite pronounced. Transmission between humans and farm animals was consequently quite rare. Both MSSA and MRSA strains belonging to otherwise pig-specific CC398 had increased capacity to colonize humans. Study of the genetic factors responsible for host specialization is underway.

Animals in an Urban Context. A Zooarchaeological study of the Medieval and Post-Medieval town of Turku

The aim of this study is to explore the role and importance of different animal species in Turku through an analysis of osteological data and documentary evidence. The osteological material used in this study is derived from two town plots in Turku dating from the 13th century to the 19th century. The osteological material deposited in Turku represents animals bred both in the town and in the surrounding landscape. Animal husbandry in SW-Finland can also be examined through a number of historical documents. The importance of animals in Turku and its hinterland are closely connected and therefore the roles of the animals in both urban and rural settings are examined. The study has revealed the complexity of the depositional patterns in medieval and post-medieval Turku. In the different areas of Turku, characteristic patterns in the osteological material and different deposit types were evident. These patterns are reflections of the activities and therefore of the lifestyles practiced in Turku. The results emphasise the importance of context- awareness in the study of material culture from archaeological sites. Both the zooarchaeological and historical sources indicate that cattle were important in animal husbandry in Turku from the Middle Ages up to the 19th century. Sheep were the second most common species. When taking into consideration the larger size of cattle, the dominance of these animals when it come to meat consumption seems clear even in those phases where sheep bones are more abundant. Pig is less abundant in the material than either cattle or sheep and their importance for subsistence was probably fairly modest, albeit constant. Goats were not abundant in the material. Most of the identified goat bones came from low utility body parts (e.g. skulls and lower extremities), but some amount of goat meat was also consumed. Wild species were of minor importance when it came to consumption practices in Turku. The changes in Turku’s animal husbandry patterns between the medieval and post medieval periods is reflected in the change in age of the animals slaughtered, which was part of a wider pattern seen in North- and Central Europe. More mature animals are also present in the assemblages. This pattern is related to the more pronounced importance of cattle as a manure producer and a draught animal as a result of the intensification of crop cultivation. This change seems to occur later in Finland than in the more Southerly regions, and indeed it did not necessarily take hold in all parts of the country.

Galactofuranose in Mycoplasma mycoides is important for membrane integrity and conceals adhesins but does not contribute to serum resistance.

Mycoplasma mycoides subsp. capri (Mmc) and subsp. mycoides (Mmm) are important ruminant pathogens worldwide causing diseases such as pleuropneumonia, mastitis and septicaemia. They express galactofuranose residues on their surface, but their role in pathogenesis has not yet been determined. The M. mycoides genomes contain up to several copies of the glf gene, which encodes an enzyme catalysing the last step in the synthesis of galactofuranose. We generated a deletion of the glf gene in a strain of Mmc using genome transplantation and tandem repeat endonuclease coupled cleavage (TREC) with yeast as an intermediary host for the genome editing. As expected, the resulting YCp1.1-Δglf strain did not produce the galactofuranose-containing glycans as shown by immunoblots and immuno-electronmicroscopy employing a galactofuranose specific monoclonal antibody. The mutant lacking galactofuranose exhibited a decreased growth rate and a significantly enhanced adhesion to small ruminant cells. The mutant was also 'leaking' as revealed by a β-galactosidase-based assay employing a membrane impermeable substrate. These findings indicate that galactofuranose-containing polysaccharides conceal adhesins and are important for membrane integrity. Unexpectedly, the mutant strain showed increased serum resistance.

Detecção de adulterações em produtos alimentares contendo leite e/ou proteínas lácteas

A critical review of the most relevant analytical methodologies for quality and authenticity control of dairy products and foods containing milk proteins is presented. Chromatographic, electrophoretic and immunological methods are used for: detection of cow's milk in ewe and goat milks, detection of whey added to milk, detection of caseins and/or whey proteins in non-lactic foods and study compounds resulting from milk proteins degradation. Techniques based on polimerase chain reaction are also suitable for detection of cow's milk on cheeses of ewe and goat milks.

Projecte de legalització d'una activitat dedicada a l'explotació ramadera i a la formatgeria artesanal

La integració en una mateixa activitat d'una explotació ramadera ecològica basada en el bestiar cabrú de llet i d'una formatgeria artesanal basada en varietats autòctones de formatge de cabra, permet ocupar un nínxol de mercat encara poc explotat actualment a Catalunya. D'igual manera, la dificultat que tenen actualment molts dels formatgers artesans del país per trobar llet de cabra en quantitats suficients i amb unes mínimes garanties higiènico-sanitàries dificulten l'avanç d'un sector amb molt bones perspectives de futur com és el de la formatgeria artesanal i justifica la necessitat de crear noves explotacions (o reconvertir-ne de ja existents) orientant-les de ple vers l'aprofitament lleter del bestiar cabrú, que és actualment molt reduït en l'àmbit Català. A més d'això, les actuals exigències en el camp del benestar animal així com en el camp de la qualitat i de la seguretat alimentaria, exigeixen a les explotacions ramaderes i a les indústries alimentàries el compliment d'una sèrie de requisits, tan de disseny dels locals i de les instal•lacions, com de control de les activitats i dels productes elaborats, que garanteixin en tot moment la seva correcte gestió i funcionament en aquestes matèries. L'objectiu d'aquest projecte, doncs, és la legalització d'una activitat, que anomenarem “Mas Peirot, S.L.”, dedicada a l'explotació ramadera ecològica de bestiar cabrú de llet i a la formatgeria artesanal de formatge de cabra. És important mencionar que tot i tractar-se d'un projecte de legalització d'una activitat,, també s'hi han contemplat aspectes i detalls relacionats amb l'execució de la mateixa, al tractar-se d'unes obres d'una certa complexitat i considerant que així es tracten amb més detall i rigorositat les actuacions projectades. En concret, en aquest projecte es planifiquen les operacions de rehabilitació de dues naus ramaderes actualment en desús, per tal que puguin destinar-se a l'explotació ramadera de bestiar cabrú i a la formatgeria artesanal, projectant-se així mateix totes les instal•lacions necessàries per tal que a les mencionades naus s'hi puguin desenvolupar les activitats citades. Com a conclusions més importants d'aquest projecte podem dir que amb un cens de 100 caps de bestiar cabrú és possible obtenir una producció mitjana de llet de 32.500 litres anuals, que es corresponen a un volum total de 3.823 quilograms de formatge a l'any (de les tipologies Formatge Garrotxa i Formatge de cabra amb oli, ambdues varietats tradicionals catalanes) obtenint un benefici de 21.389,47 Euros/any durant els primers 15 anys i un benefici de 33.386,39 Euros/any a partir del 16è any, cosa que permetrà recuperar la inversió inicial de 172.716,76 Euros que valen les naus i instal•lacions projectades, en un termini de 8 anys i permetrà, com s'ha dit, ocupar un nínxol de mercat encara poc explotat per la indústria agroalimentària a Catalunya.