Low-level Colonization Research Articles

Low-level colonization of methicillin-resistant Staphylococcus aureus in pigs is maintained by slowly evolving, closely related strains in Finnish pig farms

BackgroundOver the past two decades, livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) has become widely prevalent in pig production in Europe. The carriage status of LA-MRSA is known to vary among individual pigs, but bacterial load in pigs has rarely been studied. We assessed the quantity of LA-MRSA in nasal and skin samples of pigs and investigated the genetic diversity of the strains together with sequenced strains from national surveillance and pathology samples from the Finnish Food Authority. On two farms with assumed MRSA-positive status, farm 1 and farm 2, 10 healthy pigs were sampled three times during 2 weeks from the nares and skin (study A). On farm 1, 54 additional pigs were sampled and from confirmed MRSA-positive animals, 10 were randomly selected and transported to a clean, controlled environment for further sampling (study B). From the samples taken on farms 1 and 2 and in the controlled environment, MRSA was isolated both by direct plating and enrichment on selective media. spa types, multilocus sequence types, staphylococcal cassette chromosome mec types, resistance and virulence genes were determined. Core genome multilocus sequence typing (cgMLST) analysis was performed, including the sequences deriving from the surveillance/pathology samples from the Finnish Food Authority.ResultsAll pigs on farm 1 carried LA-MRSA in the nares at all three time points and five pigs on farm 2 at one time point. Nasal quantity varied between 10 and 103 CFU/swab and quantity on the skin between 10 and 102 CFU/swab. In the controlled environment, MRSA was detected in at least one of the nasal samples from each animal. spa type t034 was predominant. cgMLST showed one cluster with minimum allele differences between 0 and 11.ConclusionsThe study shows predominantly low-level carriage (< 103 CFU/swab) of LA-MRSA on farms. In the controlled environment we observed a decline in nasal carriage but constant skin carriage. cgMLST showed that strains of spa type t034 are closely related at the national level.

1168. Are the Staphylococcus aureus Isolates that Recolonize Infants in a NICU Following Successful Decolonization the Same or Different from the Initial Colonizing Isolate?

Abstract Background Staphylococcus aureus is an important pathogen of infants in a neonatal intensive care unit (NICU). Colonization precedes infection and decolonization may prevent infection. The origin of colonizing organisms may be the NICU environment or personnel or visitors. We have observed infants who became recolonized after successful decolonization. The purpose of this study was to determine the proportion of infants who become recolonized with the same strain or a different strain. Methods Eligible infants were consecutive infants who 1. were colonized with methicillin-susceptible S. aureus and were successfully decolonized with topical mupirocin ointment (nares and umbilicus) as evidenced by 2 or more consecutive negative weekly surveillance cultures (in the absence of a course of systemic antibiotics with activity against MSSA), 2. subsequently became recolonized, and 3. the pair of isolates was available for analysis. Isolates were analyzed by staphylococcal protein A (spa) typing and pairs with concordant spa types were subjected to whole genome sequencing (WGS; Illumina MiSeq) and phylogenetic analyses. Pairs of isolates with fewer than 25 single nucleotide polymorphism differences were considered closely related. Results There were 19 occurrences of MSSA recolonization in 17 infants following 2-6 (median, 2) negative weekly intervening surveillance cultures. Based upon spa typing (that identified 19 spa types), in 11 (58%) there was a concordant spa type and in 8 (42%) there was a discordant spa type. Of the 11 pairs of isolates with concordant spa types that were compared after WGS, 10 were closely related resulting overall in recolonization with a closely related strain in 53% of episodes. Conclusion Among MSSA colonized infants who become recolonized after successful decolonization, the recolonizing strain is the same as the original strain in over half of cases. In such cases the source is more likely to be a visitor than the NICU environment or staff. The possibility that some cases classified as recolonization were in fact persistent low level colonization or carriage in another body site not detected by surveillance cultures cannot be excluded. Disclosures Anne-Catrin Uhlemann, MD, PhD, Merck (Grant/Research Support)

Burden of perianal Staphylococcus aureus colonization in nursing home residents increases transmission to healthcare worker gowns and gloves

To evaluate the effect of the burden of Staphylococcus aureus colonization of nursing home residents on the risk of S. aureus transmission to healthcare worker (HCW) gowns and gloves. Multicenter prospective cohort study. Residents and HCWs from 13 community-based nursing homes in Maryland and Michigan. Residents were cultured for S. aureus at the anterior nares and perianal skin. The S. aureus burden was estimated by quantitative polymerase chain reaction detecting the nuc gene. HCWs wore gowns and gloves during usual care activities; gowns and gloves were swabbed and then cultured for the presence of S. aureus. In total, 403 residents were enrolled; 169 were colonized with methicillin-resistant S. aureus (MRSA) or methicillin-sensitive S. aureus (MSSA) and comprised the study population; 232 were not colonized and thus were excluded from this analysis; and 2 were withdrawn prior to being swabbed. After multivariable analysis, perianal colonization with S. aureus conferred the greatest odds for transmission to HCW gowns and gloves, and the odds increased with increasing burden of colonization: adjusted odds ratio (aOR), 2.1 (95% CI, 1.3-3.5) for low-level colonization and aOR 5.2 (95% CI, 3.1-8.7) for high level colonization. Among nursing home patients colonized with S. aureus, the risk of transmission to HCW gowns and gloves was greater from those colonized with greater quantities of S. aureus on the perianal skin. Our findings inform future infection control practices for both MRSA and MSSA in nursing homes.

Transient carriage and low-level colonization of orally administrated lytic and temperate phages in the gut of mice

Many studies have shown the efficacy of phage therapy in reducing gastrointestinal pathogens. However, it is unclear whether phages can successfully colonize the gut when administered in an adequate amount for a long time. About 1 × 108 PFU/mL of purified lytic phage PA13076 or temperate phage BP96115 were fed daily to mice via drinking water over 31 days, to elucidate the distribution of phages in the gastrointestinal tract. At day 16 and 31, six different segments of the gastrointestinal tract with their contents, including stomach, duodenum, jejunum, ileum, cecum, colon, and fresh feces, were aseptically collected. The phage titers were determined using the double-layered plate method with S. Enteritidis ATCC 13076 or S. Pullorum SPu-109 used as host cells. The results indicated that a small portion of administered phages survived exposure to gastric acid and entered the intestinal tract. The prevalence of phages in the gastrointestinal tract was lower than 1% of the primary phage count. Highest phage titers were detected in the cecum with 104 ~ 105 PFU/g, and most of the phages were eliminated from the body via feces with 106 PFU/g. On day 16 and day 31, the same level of phage titers in different segments of the gastrointestinal tract indicated that the colonization of phages had reached saturation at day 16. These results demonstrate transient phage carriage and low-level colonization of orally administrated lytic and temperate gut phages in mice.Graphical abstract

Propionibacterium acnes Induces Autophagy in Keratinocytes: Involvement of Multiple Mechanisms

Propionibacterium acnes is a dominant member of the cutaneous microbiota. Herein, we evaluate the effects of different P.acnes strains and propionic acid on autophagy in keratinocytes. Our results showed that P.acnes strain 889 altered the architecture of the mitochondrial network; elevated the levels of microtubule-associated protein 1 light chain 3B-II, Beclin-1, and phospho-5'-adenosine-monophosphate-activated protein kinase α; stimulated autophagic flux; facilitated intracellular redistribution of microtubule-associated protein 1 light chain 3B; increased average number of autophagosomes per cell; and enhanced development of acidic vesicular organelles in the HPV-KER cell line. Propionic acid increased the level of phospho-5'-adenosine-monophosphate-activated protein kinase α, enhanced lipidation of microtubule-associated protein 1 light chain 3B, stimulated autophagic flux, and facilitated translocation of microtubule-associated protein 1 light chain 3B into autophagosomes in HPV-KER cells. P.acnes strains 889 and 6609 and heat-killed strain 889 also stimulated autophagosome formation in primary keratinocytes to varying degrees. These results indicate that cell wall components and secreted propionic acid metabolite of P.acnes evoke mitochondrial damage successively, thereby triggering 5'-adenosine-monophosphate-activated protein kinase-associated activation of autophagy, which in turn facilitates the removal of dysfunctional mitochondria and promotes survival of keratinocytes. Thus, we suggest that low-level colonization of hair follicles with noninvasive P.acnes strains, by triggering a local increase in autophagic activity, might exert a profound effect on several physiological processes responsible for the maintenance of skin tissue homeostasis.

CampylobacterInfection Has Different Outcomes in Fast- and Slow-Growing Broiler Chickens

Campylobacter spp. are frequently carried by poultry, but they are not believed to cause significant disease in these animals. Modern poultry breeds have been selected to grow rapidly under intensive conditions, but recently, consumers have moved toward purchasing birds produced in higher welfare, free-range or organic systems. Birds reared in these systems tend to be a slower growing breed and are fed a different diet. Birds reared in such systems are stocked at a lower density compared with the standard conventional broilers, and they have access to environmental enrichment, such as perches. In previous research, these slower growing birds have been shown to have different levels of Campylobacter carriage in commercial rearing conditions, but the reasons for, and effect of, these different levels are unknown; is it the bird breed, diet, or environmental conditions? In this study, experimental flocks of fast- and slow-growing breeds of broiler chickens were reared to a standard commercial slaughter weight, with their weight gain being measured during the growing period. At 21 days, birds were either infected with Campylobacter jejuni or given a placebo as control. Cohorts of birds were euthanatized at various intervals, and samples were taken for examination for Campylobacter. The fast-growing birds gained weight more rapidly than the slow-growing birds. By 2 days postinfection (dpi), C. jejuni was detected in the caeca and by enrichment from the liver and spleen samples from both breeds of birds. Low-level colonization persisted in the spleen and liver samples but was undetectable by 28 dpi. Fast- and slow-growing birds did not show detectably different levels of Campylobacter carriage. Infection with C. jejuni affected the incidence of hock marks and pododermatitis in both breeds of birds, but the differences were greater with the fast-growing breed compared with the uninfected control birds. In addition, the incidence of pododermatitis was significantly higher in Campylobacter-positive fast-growing birds than in their slower-growing counterparts. The results show that infection with Campylobacter can have an indirect welfare effect on birds via increased incidence of hock marks and pododermatitis.

Disappearing Microbiota: Helicobacter pylori Protection against Esophageal Adenocarcinoma

Once rare, esophageal adenocarcinoma (EAC) is the most rapidly increasing malignancy in many developed countries, such as the United States, United Kingdom, Australia, and Norway, and now is surpassing esophageal squamous cell carcinomas in certain populations ([1][1], [2][2]). EAC has very specific

The GacA global regulator of Vibrio fischeri is required for normal host tissue responses that limit subsequent bacterial colonization

Harmful and beneficial bacterium-host interactions induce similar host-tissue changes that lead to contrasting outcomes of association. A life-long association between Vibrio fischeri and the light organ of its host Euprymna scolopes begins when the squid collects bacteria from the surrounding seawater using mucus secreted from ciliated epithelial appendages. Following colonization, the bacterium causes changes in host tissue including cessation of mucus shedding, and apoptosis and regression of the appendages that may limit additional bacterial interactions. We evaluated whether delivery of morphogenic signals is influenced by GacA, a virulence regulator in pathogens, which also influences squid-colonization by V. fischeri. Low-level colonization by a GacA mutant led to regression of the ciliated appendages. However, the GacA mutant did not induce cessation of mucus shedding, nor did it trigger apoptosis in the appendages, a phenotype that normally correlates with their regression. Because apoptosis is triggered by lipopolysaccharide, we examined the GacA mutant and determined that it had an altered lipopolysaccharide profile as well as an increased sensitivity to detergents. GacA-mutant-colonized animals were highly susceptible to invasion by secondary colonizers, suggesting that the GacA mutant's inability to signal the full programme of light-organ responses permitted the prolonged recruitment of additional symbionts.

Worms, germs and IBD: Is it your mother’s fault?

Worms, germs and IBD: Is it your mother’s fault?

Low-level colonization and infection with ciprofloxacin-resistant gram-negative bacilli in a skilled nursing facility

Background: We report a 1-year surveillance study that evaluates colonization and infection with ciprofloxacin-resistant gram-negative bacilli (CR GNB) and the relation to quinolone use and other possible risk factors in a proprietary skilled nursing facility (SNF) with no history of outbreaks. Methods: Rectal swabs obtained quarterly were streaked on MacConkey agar with ciprofloxacin discs (5 μg) to screen for CR GNB and later were speciated and the antimicrobial susceptibilities were confirmed by standardized disc-diffusion tests. Results: The mean prevalence of CR GNB colonization was 2.6% (range 0.9% to 5.3%). The colonization frequency was higher in the last survey than it was in the first survey. CR GNB–colonized strains included Pseudomonas species (21%), but more than half were non-Pseudomonas enterics such as Acinetobacter baumannii (25%), Proteus mirabilis (17%), and Providencia stuartii (13%). None of the patients who had colonization with CR GNB had subsequent infections with the same species. Patients with colonization had more exposure to ciprofloxacin and they were more likely to have been recently admitted from an acute-care hospital and have decubitus ulcers. During the study period, of 336 patients surveyed, 98 (29%) patients developed suspected infections and cultures were done; the infection rate was 4.7 per 1000 patient days. Of these infected patients, 59 (60%) were infected by GNBs; the infection rate was 2.3 per 1000 patient days. Nineteen percent of the GNB infections were treated with a quinolone. (Overall, quinolones constituted about 17% of antibiotic usage in the SNF). Only 3 (5%) of the patients infected with GNB were infected with CR GNB, including Pseudomonas and Providenci a species. The CR GNB infections involved multiple sites, multiple organisms, and long length of stay in the SNF. Conclusions: The findings indicate that in this community SNF, a low frequency of colonization or infection with CR GNB existed. Whether continued moderate use of quinolones will lead to increasing levels of CR GNB will require further study. (AJIC Am J Infect Control 1998;26:552-7)

Mycorrhizal colonization and nutrient uptake of dry bean in manure and compost manure treated subsoil and untreated topsoil and subsoil

Eroded or leveled Portneuf silt loam soils (coarse‐silty mixed mesic Durixerollic Calciorthid) have been restored to topsoil productivity levels by manure application, but not by other organic sources such as cheese whey. In dry bean (Phaseolus vulgaris L. cv. Viva), only soil organic matter and Zn concentration of leaf tissue correlated with improved yields. Manure application could potentially increase or decrease mycorrhizal colonization depending on which factors dominate. Manured and unmanured soils from a long‐term field experiment were sampled and mycorrhizal spores were quantified, but there was no significant manure treatment effect on spore numbers. A greenhouse study was conducted to see if manure or composted manure freshly applied to subsoils would facilitate mycorrhizal colonization in dry bean roots compared to untreated topsoil or conventionally fertilized subsoil. Low level colonization (< 5%) was observed 21 days after planting and that increased to 58% by 56 days after planting. Roots grown on subsoil treated with manure or composted manure showed higher percent colonization than roots from untreated subsoil, but roots on topsoil had highest colonization. This increase in colonization was statistically significant for the last two sampling dates. Topsoil promoted the greatest percent colonization in early bean growth and this was reflected in greater Zn uptake during early growth stages. By day 56, plants grown in manured subsoil absorbed Zn equal to topsoil and at higher levels than the subsoil control. However, this increase in Zn uptake was not seen in plants grown in compost manured subsoil. A decrease in root and shoot weight was observed in the composted manure treatment and this seemed to decrease mycorrhizal efficiency. Uptake of other nutrients was either not related or was negatively related to mycorrhizal infection. The higher percent colonization of roots by mycorrhizal fungi stimulated by manure could explain the field observations of higher bean yield and Zn contents in dry bean in manured than in untreated subsoils.

The effect of selective decontamination of the digestive tract with the addition of systemic cefotaxime on the aerobic faecal flora of mice.

The administration per-orally to mice of the non-absorbable antibiotics polymyxin E, tobramycin and amphotericin B resulted in the elimination of detectable aerobic gram-negative rods from the faecal flora without affecting the total viable aerobic count. The addition of parental cefotaxime to the regime caused a fall in the number of aerobic lactobacilli and an increase in the number of enterococci. The rise was associated with the translocation of viable enterococci to the mesenteric lymph nodes and the spleen. The changes induced by cefotaxime were reversed when the antibiotic was withdrawn. Following withdrawal of all antibiotics the total aerobic faecal flora increased to above normal levels, but there was no associated diarrhoea. Attempts to implant exogenous enterobacteria into the digestive tract resulted in only low level colonization both in treated mice and in control mice. These results may have implications for the use of this antibiotic regime for selective decontamination of the digestive tract in humans, particularly those who are immunocompromised.