Penicillin-binding protein gene alterations in Streptococcus uberis isolates presenting decreased susceptibility to penicillin.

Streptococcus uberis is an environmental pathogen commonly causing bovine mastitis, an infection that is generally treated with penicillin G. No field case of true penicillin-resistant S. uberis (MIC > 16 mg/liter) has been described yet, but isolates presenting decreased susceptibility (MIC of 0.25 to 0.5 mg/liter) to this drug are regularly reported to our laboratory. In this study, we demonstrated that S. uberis can readily develop penicillin resistance in laboratory-evolved mutants. The molecular mechanism of resistance (acquisition of mutations in penicillin-binding protein 1A [PBP1A], PBP2B, and PBP2X) was generally similar to that of all other penicillin-resistant streptococci described so far. In addition, it was also specific to S. uberis in that independent resistant mutants carried a unique set of seven consensus mutations, of which only one (Q(554)E in PBP2X) was commonly found in other streptococci. In parallel, independent isolates from bovine mastitis with different geographical origins (France, Holland, and Switzerland) and presenting a decreased susceptibility to penicillin were characterized. No mosaic PBPs were detected, but they all presented mutations identical to the one found in the laboratory-evolved mutants. This indicates that penicillin resistance development in S. uberis might follow a stringent pathway that would explain, in addition to the ecological niche of this pathogen, why naturally occurring resistances are still rare. In addition, this study shows that there is a reservoir of mutated PBPs in animals, which might be exchanged with other streptococci, such as Streptococcus agalactiae, that could potentially be transmitted to humans.

Staphylococcus aureus host range and human-bovine host shift.

Staphylococcus aureus is a major agent of bovine mastitis. The concomitant emergence of pig-associated methicillin-resistant S. aureus (MRSA) in human carriage and infection requires a reexamination of the host range and specificity of human- and cow-associated S. aureus strains, something which has not been systematically studied previously. The genetic relatedness of 500 S. aureus isolates from bovine mastitis cases, 57 isolates from nasal carriage of farmers, and 133 isolates from nonfarmers was determined by amplified fragment length polymorphism (AFLP) analysis and spa typing. Multilocus sequence typing (MLST) was conducted on a subset of isolates to match AFLP clusters with MLST clonal complexes (CCs). This data set allowed us to study host range and host specificity and to estimate the extent of bovine-to-human transmission. The genotype compositions of S. aureus isolates from farmers and nonfarmers were very similar, while the mastitis isolates were quite distinct. Overall, transmission was low, but specific genotypes did show increased cow-to-human transmission. Unexpectedly, more than one-third of mastitis isolates belonged to CC8, a lineage which has not been considered to be bovine mastitis associated, but it is well known from human carriage and infection (i.e., USA300). Despite the fact that we did detect some transmission of other genotypes from cows to farmers, no transmission of CC8 isolates to farmers was detected, except for one tentative case. This was despite the close genetic relatedness of mastitis CC8 strains to nonfarmer carriage strains. These results suggest that the emergence of the new bovine-adapted genotype was due to a recent host shift from humans to cows concurrent with a loss of the ability to colonize humans. More broadly, our results indicate that host specificity is a lineage-specific trait that can rapidly evolve.

Kyselytutkimus eläinlääkärien kokemuksista EELAn valmistamien autogeeni- ja varastostafylokokkitoksoidirokotteiden tehosta ja turvallisuudesta lehmän mastiitin vastustamisessa

Summary: Questionnaire survey concerning the experience of veterinarians about the efficacy and safety of the autogenous inactivated staphylococcal toxoid vaccine used against mastitis in cattle in Finland

Naudan kliinisen utaretulehduksen bakteerietiologia : Saaren ambulatorisen klinikan aineisto vuosilta 2002-2003

Summary: Bacterial etiology of bovine clinical mastitis - data from Saari Ambulatory Clinic in 2002-2003

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

09006-003 Bear Creek Watershed Final Report

Bear Creek is an impaired warm water fishery designated as class B(LR) by the Iowa DNR and is on 303 impaired waters list for fish kills and ammonia. Bear Creek is located in eastern Delaware County. This project is designed to improve the water quality of Bear Creek by educating the landowners, operators and watershed community about the importance of this water resource. The goal of the Bear Creek Watershed Project is to improve the water quality of Bear Creek by reducing the amounts of ammoniated manure discharge, fecal coliform bacteria, sediment, nitrogen, and phosphorous. The Bear Creek Watershed Project has been a watershed project since July 2004, first as a Demo project FY 2004-2005 and then full time WSPF/319 project FY06-09. Fish kills have not occurred in 2008-2009. Sediment delivery has decreased in the Bear Creek Watershed by 5,328 tons per year. The objectives of this watershed project will be to improve Livestock Waste Storage, to improve Livestock Waste Usage, to decrease Sediment Losses, and to improve Education & Area Outreach. This project will install 2 manure storage structures (EQIP/project funded), 19 ac of CRP waterways, 12 ac of project waterways, 17 ac of CRP filter strips along stream, 12 water and sediment control basins, 18,000 ft of terraces, 350 ac of new notill planting, and 3,700 ft of streambank protection.

07031-011 Brushy Creek Watershed Final Report

Brushy Creek is a tributary of the Raccoon River, which is a regular source of drinking water for over 400,000 Iowans. Regular monitoring by Des Moines Water Works (DMWW) and Agriculture’s Clean Water Alliance (ACWA) over the last eight years has shown the stream to be highly impaired for coliform bacteria and nitrate. Both Brushy Creek and the Raccoon River are on the 303(d) impaired waterbody list. A December 2005 fish kill in Brushy Creek resulted in administrative actions against seven livestock producers. Several open feed lots exist in the watershed. The community of Roselle (in the Brushy Creek watershed) has been identified by IDNR as unsewered, and many dwellings throughout the watershed discharge untreated human waste. No Watershed Improvement Association (WIA) exists in this sparsely-populated area. This outcome-based project will: • Enhance nutrient and manure management to reduce agricultural inputs to the stream. • Assess the amount of human waste reaching the stream from Roselle. • Engage and inform local residents so a WIA can be formed. • Monitor performance through a rigorous water and soil testing program. This project embraces a concept of participation from all levels of government, commodity organizations, and the private sector. The largest drinking water utility in the state will lead and administer this effort. The participating parties will work to establish a functioning WIA so that progress achieved through this project will be robust and long-lasting. The participants believe this will be the most effective approach to correct the situation, and will serve as a model for other problem watersheds throughout the state.

06031-014 City of Carpenter Sewage Treatment System Project Final Report

The Mitchell County Soil and Water Conservation District is applying on behalf of the incorporated community of Carpenter to construct a wastewater collection and treatment system to assist in the environmental cleanup and protection of Deer Creek. IDNR water monitoring of the community tile line has shown consistently elevated levels of fecal coliform bacteria indicating the presence of untreated sewage water. These are obvious health threats to the downstream users and wildlife in Deer Creek and the Cedar River. A new sewer system for the community of Carpenter will eliminate illegal discharges into the creed and be the first step in the overall protection of the stream.

05011-055 Yellow River Watershed Improvement Project Final Report

An overall effort has been initiated to improve the quality of the Yellow River in Northeast Iowa by reducing the amount of sediment and bacteria entering the stream. Funding for this project will be utilized to improve stream quality to the level of fully supporting game fish such as brown, rainbow and brook trout, walleye, northern pike and smallmouth bass. The Yellow River has the potential to be one of the top trout streams, not only in Iowa, but in the entire Upper Midwest. This project will greatly enhance recreational activities such as fishing, canoeing and inner tubing and will greatly increase tourism dollars to the state. The project will specifically address two sources of impairment: stream bank erosion and coliform bacteria from both livestock and inadequate human septic systems.

Klebsiella utaretulehduksen aiheuttajana : kirjallisuuskatsaus ja tapausselostus

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

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.