Using high-resolution imaging to study the impact of the Kalogon wheelchair cushion on blood flow in the gluteal area.

AIM OF THE STUDY: This study investigated how the air-bladder offloading mode of the Orbiter by Kalogon wheelchair cushion (Orbiter) affected blood flow in the gluteal region of non-disabled subjects. The hypothesis was that the cushion's offloading mode would improve blood flow, resulting in reduced reactive hyperemia when compared to the static setting, or Loaded Control (LC). Furthermore, the study proposed a technique using a high-resolution image laser speckle contrast system to measure blood flow in the gluteal area. METHODS: Two procedures were carried out, one with the participant sitting on a cushion in LC, and the second, the cushion was set to offloading mode. Blood flow was measured through data imaging after each procedure. Three trials were performed, starting and ending in different cushion bladders. Customized algorithms were used to select regions of interest on the images for calculations. The Wilcoxon Signed-Rank Test was conducted to compare the offloads and loaded control values of each region of interest. Results were considered significant at α = 0.05. RESULTS: Ten healthy, non-disabled adults participated in the study, seven females and three males. There were no significant differences among the participants. However, results showed that seven subjects tended to decrease reactive hyperemia in the offload sequence of trial when the last two bladders offloaded were the sacrum followed by the right ischial tuberosity. CONCLUSIONS: The high-resolution imager showed that the Orbiter Offloads helped reduce reactive hyperemia in seven subjects, potentially improving blood flow. More research is necessary to comprehend the mechanisms of these effects fully.

Healthcare-Associated Infections and the Hospital Bed.

OBJECTIVE: Bedframes are a potential source of bacterial contamination, fomites, and healthcare-associated infections for patients with active skin wounds and other underlying conditions. Bedframes also differ in their design, materials, texture, and ease of disassembly for cleaning. In this study, the authors evaluated five hospital bedframes in terms of retained soil and ease of cleaning as rated by volunteers. METHODS: Hospital mattresses were placed on five different bedframes and soiled with mock bodily fluids containing Geobacillus stearothermophilus endospores as an indicator organism for contamination. In a second set of experiments, volunteers evaluated the bedframes for ease of cleaning; fewer than 30% of the volunteers had experience cleaning in hospitals or had previously received infection-control training. Questionnaires evaluated subjective measures such as ease of cleaning and texture. RESULTS: Researchers observed a strong correlation between the initial amount of soil retained, the most probable number calculations of endospore counts, and the number of washes to reach extinction (no detectable endospores). Although volunteers' rankings for ease of cleaning were independent of the amount of soil retained, their rankings correlated with the actual washes to reach undetectable limits and bedframe materials that were perceived as harder to clean. CONCLUSIONS: This study demonstrates the importance of both bedframe design and user cleaning experience in reducing bedframes as a source of healthcare-associated infections.

Comparing the Energy-Stretch Properties of Two Compression Bandage Systems in a Laboratory-Based Test under Controlled Conditions.

OBJECTIVE: To compare the characteristics of two commercially available compression systems, a dual-compression bandage system (DCS) and a traditional two-layer bandage (TLB), using a laboratory bench test. METHODS: The compression systems were evaluated in a computer-controlled tensile test to generate force-deflection curves for each sample. The compressive work and the theoretical pressure applied to the limb by the respective compression bandages were calculated at the maximum stretch and a stretch instructed by the manufacturers. The manufacturer of the DCS provides reference points on how much the bandage should be stretched to provide the desired pressure, and the TLB stretch was calculated from the product's datasheet. RESULTS: The combined results of layers 1 and 2 for the DCS showed greater load and work than the TLB at both the maximum and recommended stretch. The recommended stretch for DCS and TLB was less than 50% of the deflection up to the breaking point. CONCLUSIONS: The high work provided by the two layers of the DCS suggests a wider range of performance than the TLB when applied to the lower limb, especially after the limb volume is initially reduced by compression. Moreover, using the tensile test and the guide of the reference points on layers 1 and 2 from DCS, the calculated pressure achieved the expected values stated by the manufacturer. Human studies should be conducted to determine whether the reference points provided by DCS are beneficial for obtaining repeatable values.

Invited review: Review of taxonomic changes in dairy-related lactobacilli.

The genus Lactobacillus has represented an extremely large and diverse collection of bacteria that populate a wide range of habitats, and which may have industrial applications. Researchers have grappled with the immense genetic, metabolic, and ecological diversity within the genus Lactobacillus for many years. As a result, the taxonomy of lactobacilli has been extensively revised, incorporating new genus names for many lactobacilli based on their characteristics including genomic similarities. As a result, many lactobacilli traditionally associated with dairy products now have new genus names and are grouped into new clades or clusters of species. In this review, we examine how the taxonomic restructuring of the genus Lactobacillus will affect the dairy industry and discuss lactobacilli associated with dairy production, processing, and those that confer possible health benefits when delivered by dairy products.

Comparison of growth and survival of single strains of Lactococcus lactis and Lactococcus cremoris during Cheddar cheese manufacture.

Traditionally, starter cultures for Cheddar cheese are combinations of Lactococcus lactis and Lactococcus cremoris. Our goal was to compare growth and survival of individual strains during cheesemaking, and after salting and pressing. Cultures used were 2 strains of L. lactis (SSM 7605, SSM 7436) and 2 strains of L. cremoris (SSM 7136, SSM 7661). A standardized Cheddar cheese make procedure was used that included a 38°C cook temperature and salting levels of 2.0, 2.4, 2.8, 3.2, and 3.6% from which were selected cheeses with salt-in-moisture levels of 3.5, 4.5, and 5.5%. Vats of cheese were made using each strain on its own as biological duplicates on different days. Starter culture numbers were enumerated by plate counting during cheesemaking and after 6 d storage at 6°C. Flow cytometry with fluorescent staining by SYBR Green and propidium iodide was used to determine the number of live and dead cells in cheese at the different salt levels. Differences in cheese make times were strain dependent rather than species dependent. Even with correction for average culture chain length, cheeses made using L. lactis strains contained ∼4 times (∼0.6 log) more bacterial cells than those made using L. cremoris strains. Growth of the strains used in this study was not influenced by the amount of salt added to the curd. The higher pH of cheeses with higher salting levels was attributed to those cheeses having a lower moisture content. Based on flow cytometry, ∼5% of the total starter culture cells in the cheese were dead after 6 d of storage. Another 3 to 19% of the cells were designated as being live, but semipermeable, with L. cremoris strains having the higher number of semipermeable cells.

Galactose-positive adjunct cultures prevent gas formation by Paucilactobacillus wasatchensis WDC04 in a model gas production test.

Gas production by obligatory heterofermentative lactic acid bacteria such as Paucilactobacillus wasatchensis is a sporadic problem in Cheddar cheese and results in undesired slits and cracks in the cheese. Growth of Pa. wasatchensis is not rapid, which makes investigations of gas production difficult to consistently execute. A primary objective of this study was to develop a model gas production test that could be used to investigate the effect of galactose and ribose utilization on gas production by Pa. wasatchensis and determine whether galactose-fermenting adjunct cultures could prevent gas formation. Paucilactobacillus wasatchensis WDC04 was inoculated at 101 to 106 cfu/mL into carbohydrate-restricted MRS broth containing different ribose and galactose levels and incubated for up to 21 d at 23°C. Gas production in the broth was detected using a Durham tube inverted on a 6-cm-long capillary tube; cells were enumerated at 4, 8, and 12 d; and residual galactose was also measured. Gas production was sporadic except for when 105 cfu/mL of Pa. wasatchensis WDC04 was inoculated into broth containing 0.3% ribose and 0.7% galactose. In those tubes, gas production was consistently observed after 8-d incubation, by which time galactose levels had decreased to 0.15%. Co-inoculation of Pa. wasatchensis WDC04 with as few as 103 cfu/mL of a lactose-negative galactose-positive adjunct culture (Pediococcus acidilactici 23F, Lacticaseibacillus paracasei UW4, or Lactobacillus helveticus 7995) resulted in galactose depletion by d 4 and no observable gas production by d 12. With less galactose available to the slower-growing Pa. wasatchensis WDC04, its growth was limited to 108 cfu/mL when any of the adjunct cultures was co-inoculated, compared with 109 cfu/mL when grown on its own. We concluded that galactose-fermenting adjunct cultures have potential for preventing unwanted gas production in cheese by competition for resources and especially by removing the 6-carbon galactose before it can be utilized for energy by an obligatory heterofermentative lactobacilli such as Pa. wasatchensis and produce carbon dioxide.

Gluconate metabolism and gas production by Paucilactobacillus wasatchensis WDC04.

Paucilactobacillus wasatchensis, a nonstarter lactic acid bacteria, can cause late gas production and splits and cracks in aging cheese when it metabolizes 6-carbon substrates, particularly galactose, to a 5-carbon sugar, resulting in the release of CO2. Previous studies have not explained late gas production in aging cheese when no galactose is present. Based on the genome sequence of Pa. wasatchensis WDC04, genes for potential metabolic pathways were mapped using knowledgebase predictive biology software. This metabolic modeling predicted Pa. wasatchensis WDC04 could metabolize gluconate. Gluconate contains 6 carbons, and Pa. wasatchensis WDC04 contains genes to convert it to 6-P-gluconate and then to ribulose-5-P by using 6-phosphogluconate dehydrogenase in a decarboxylating step, producing CO2 during its metabolism. The goal of this study was to determine if sodium gluconate, often added to cheese to reduce calcium lactate crystal formation, could be metabolized by Pa. wasatchensis WDC04, resulting in gas production. Carbohydrate-restricted DeMan, Rogosa, and Sharpe broth was mixed with varying ratios of ribose, sodium gluconate, or d-galactose (total added substrate content of 1% wt/vol). Oxyrase (Oxyrase Inc.; 1.8% vol/vol) was also used to mimic the anaerobic environment of cheese aging in selected tubes. Tubes were inoculated with a 4-d culture of Pa. wasatchensis WDCO4, and results were recorded over 8 d. When inoculated into carbohydrate-restricted DeMan, Rogosa, and Sharpe broth containing only sodium gluconate as the added substrate, Pa. wasatchensis WDC04 grew, confirming gluconate utilization. Of the 10 ratios used, Pa. wasatchensis WDC04 produced gas in 6 scenarios, with the most gas production resulting from the ratio of 100% sodium gluconate with no added ribose or galactose. It was confirmed that obligately heterofermentative nonstarter lactobacilli such as Pa. wasatchensis WDC04 can utilize sodium gluconate to produce CO2 gas. Addition of sodium gluconate to cheese thus becomes another risk factor for unwanted gas production and formation of slits and cracks.

Keeping Patients Under the Damage Threshold for Pressure Injury: Addressing Microclimate Through Allostasis.

BACKGROUND: Pressure injuries (PI) are an ongoing problem in health care. Current interventions, both from clinicians and support surface technologies, do not sufficiently address PI prevention. PROBLEM: Patient microclimate is a contributing risk factor for PI, one which can be more adequately addressed. However, the acceptable range for microclimate is unknown, in part because the body adapts to changing conditions. APPROACH: Two key concepts in allostasis are finite resources and responding to intrinsic and extrinsic demands. These concepts have not previously been applied to PI treatment or interventions. Addressing microclimate, when coupled with an increasing awareness of the cumulative effect of individual patient risk factors, can help resolve the risk of PI by lowering the cumulative inputs to keep patients under the threshold for tissue damage. CONCLUSION: This new approach, which places microclimate risk into the broader conceptual framework of allostasis, can produce more effective products and interventions to prevent PI.

Use of a Support Surface Standard to Test the Effects of a Turning and Positioning Device Versus Low-Air-Loss Therapy on Temperature and Humidity.

BACKGROUND: Turning and repositioning devices (TRDs) help to reduce strain on caregivers, but clinicians question their effects on humidity and temperature (microclimate) at the skin surface that may increase risk of pressure ulcers. OBJECTIVE: To pilot the use of a standard test for support surfaces to compare microclimate at the skin surface in three scenarios: (1) on a low-air-loss (LAL) surface, (2) on a representative TRD with a basic underpad (TRDU) placed on a LAL surface, and (3) on a negative control with full occlusion. The results are designed to inform clinical decision-making in using a TRD on a LAL surface and the viability of using this test to study TRDs. DESIGN: Measuring humidity and temperature at the device-surface interface using a heated moisture-exuding bronze thermodynamic human model in a laboratory setting. MAIN OUTCOME MEASURE: Humidity and temperature levels across 3 hours 15 minutes of continuous loading with a 45-second complete unloading to simulate a position change at 3 hours. MAIN RESULTS: Relative humidity on the TRDU was below that on the LAL surface for the first 110 minutes and was markedly lower than the negative control for the remainder of humidity testing. Temperature on the TRDU was well below the negative control and negligibly higher than the surface alone throughout testing. The position change enhanced the effects of the TRDU. CONCLUSIONS: The support surface standard test appears useful in evaluating TRDs. This TRD along with the basic underpad is more comparable to a LAL surface than to full occlusion in managing the microclimate of the skin and pressure ulcer risk.

Results of Laboratory Testing for Immersion, Envelopment, and Horizontal Stiffness on Turn and Position Devices to Manage Pressure Injury.

BACKGROUND: A continuing complication, pressure injuries are due to sustained mechanical loading and tissue deformations, which can then be exacerbated by additional intrinsic and extrinsic risk factors. Although support surfaces are designed to mitigate risk factors for pressure injuries, the presence of a turn and position device (TPD) between the patient and support surface may interfere with how support surfaces affect these risk factors. OBJECTIVE: Report the use of the NPIAP's S3I standard test methods to characterize the performance of a support surface when used in conjunction with three different TPDs. DESIGN: Laboratory testing compared three TPDs for Immersion, Envelopment, and Horizontal Stiffness in each of five surface combinations. MAIN OUTCOME MEASURE: Immersion test measures how far mannequin indenter immerses into surface. Envelopment test measures immersion and pressure distribution with hemispherical-indenter with mounted sensor rings. Horizontal Stiffness test measures the shear modulus of the support surface with epicondyle indenter. MAIN RESULTS: For the specific TPDs tested here, the one with an adjustable integrated air bladder improved rather than compromised both the envelopment and the immersion of the support surface alone. Additionally, this TPD provided potential protection against sliding and the associated frictional shear forces. CONCLUSIONS: This paper describes how TPDs should perform in order to help establish which features are needed in a new medical device of this type. Laboratory testing demonstrates it is possible to improve performance of a support surface by applying a TPD as an add-on, thus relieving tissue deformation exposure through more effective pressure redistribution.

Growth and survival characteristics of Paucilactobacillus wasatchensis WDCO4.

Understanding characteristics that permit survival and growth of Paucilactobacillus wasatchensis as part of the nonstarter microbiota of cheese is important for minimizing unwanted gas formation in cheese that can cause downgrading because of slits and cracks. The ability of Plb. wasatchensis WDC04 to survive pasteurization was studied by inoculating raw milk with 108 cfu/mL and measuring survival after processing through a high-temperature, short-time pasteurizer. Extent and rate of growth of Plb. wasatchensis WDC04 as a function of pH, salt concentration, and presence of various organic acids were studied using 48-well microplates in an automated spectrophotometer measuring optical density at 600 nm. Better growth in the 1-mL wells was obtained when a micro-anaerobic environment (similar to that which occurs in cheese) was created by enzymically removing the oxygen. Faster growth occurred around neutral pH (pH 6 to 8) than at pH 5 (cheese pH), whereas only marginal growth occurred at pH 4. Adding sodium chloride retarded growth of Plb. wasatchensis WDC04, but slow growth occurred even at salt concentrations up to 6%. At salt-in-moisture (S/M) concentrations found in cheese, the rate of growth at 3.5% S/M >4.5% S/M >5.5% S/M. Thus, low salt level in cheese is a risk factor for Plb. wasatchensis growth during cheese storage and unwanted slits and cracks. Some of the organic acids tested (propionic, formic, and citric) tended to suppress growth of Plb. wasatchensis WDC04 more than would be expected from their effect on pH. No survival of Plb. wasatchensis WDC04 after pasteurization was observed with the reduction in numbers being 8 logs or more. Even subpasteurization heating at 69°C for 15 s was sufficient to inactivate Plb. wasatchensis WDC04, so its presence as part of the nonstarter microbiota of cheese should be considered as a postpasteurization environmental contamination.

New Clinically Relevant Method to Evaluate the Life Span of Prophylactic Sacral Dressings.

It has been demonstrated that wound dressings provide a protective effect against pressure injuries. However, no method exists to measure either the life or performance of dressings used in prevention; testing dressings in a clinical setting or a research environment has typically been based on measuring its moisture absorption capacity. This article examines the changes that occur in the structural and mechanical properties of a prophylactic dressing based on conditions of use when wound exudate is not present.A clinically relevant method was developed to simulate the loading, friction-inducing shear, and moisture transpiration present in a typical hospitalization where a dressing is applied for prevention. Single-use dressings were tested using this method to evaluate their ability to protect patients from pressure injuries throughout the typical 5 to 7 days of use. Following this aging process, researchers measured the physical, structural, and mechanical changes in prophylactic dressings over time.This innovative method provides guidance for clinicians on dressing use and replacement intervals. For bioengineers, the method generates important empirical data for computer modeling of dressing performance, which can then reveal the consequences of changes in dressing structure and function on sustained tissue loads. It is the authors' hope to generate discussion about the creation of industry-wide standards for testing dressings to improve patient care.

Measuring Tensile Strength to Better Establish Protective Capacity of Sacral Prophylactic Dressings Over 7 Days of Laboratory Aging.

Results from large-scale randomized clinical trials support the application of prophylactic dressings to provide protection from body-weight force-induced deformations known to damage skin and underlying tissues, which often result in pressure injuries (pressure ulcers). This laboratory study using a new method for aging dressings in simulated use followed by tensile testing was conducted to further understand the protective effect of sacral prophylactic dressings (SPDs) in alleviating tissue deformations in the sacral region through the course of typical application. Specifically, four SPDs were exposed to a simulation of the clinical environment incorporating saline solution absorption, mechanical loading, and repetitive sliding-induced shear. After aging, the protective endurance of the SPDs was measured through tensile testing to determine their effectiveness against tissue-damaging forces over time.This study uses the concepts of axial stiffness, protective endurance, and elastic limit to describe more accurately the protective aspects of SPDs under dry and moist conditions and how they interact with the skin and underlying tissues over the life of the dressing. The authors propose two primary features in SPD effectiveness in preventing pressure injuries: high conformability (ie, low flexural stiffness) and protective endurance (the dressing's capacity to maintain biomechanical performance when moist).

Hazardous Drug Contamination of Drug Preparation Devices and Staff: A Contamination Study Simulating the Use of Chemotherapy Drugs in a Clinical Setting.

Background: Hazardous drug contamination (HD) in healthcare environments continues, placing healthcare staff at risk of significantly chronic health problems, despite the use of personal protective equipment (PPE) and closed system transfer device systems (CSTDs). Objective: This study's aim was to determine how HD might spread through touch after handling contaminated vials in simulated pharmacy and nursing environments. Methods: UV fluorescent powder Glo Germ (Glo Germ Co., Moab, UT), composed of melamine resin, was used to simulate HD. Following manufacturer's exact usage specifications, five commercially available CSTDs were tested in the simulated pharmaceutical preparation environment under controlled conditions. UV fluorescence was used to detect the test medium powder that simulated HD following each trial. Photographs, using a Nikon D40X (10.2 mp) camera, were taken at selected stages of testing to document the presence of HD. Results: Transfer of the HD testing medium (Glo Germ) to IV sets, pharmacy PPE, and nursing PPE was observed in 4 of 5 CSTDs tested. The only CSTDs that showed no observable contamination was the Allison Medical Safety Enclosed Vial Adapter (SEVA) system (Littleton, CO). Conclusions: Results show residue from HD vials spread as vial was handled in a pharmacy environment in 4 of 5 CSTDs, contaminating pharmacy PPE, ancillary products, and nursing PPE. No HD residue was detected on PPE, ancillary products, or nursing PPE in the pharmacy after using the SEVA system, providing an effective means to contain HD for drug transfer from vial to delivery system.

Hot topic: Geographical distribution and strain diversity of Lactobacillus wasatchensis isolated from cheese with unwanted gas formation.

Lactobacillus wasatchensis, an obligate heterofermentative nonstarter lactic acid bacteria (NSLAB) implicated in causing gas defects in aged cheeses, was originally isolated from an aged Cheddar produced in Logan, Utah. To determine the geographical distribution of this organism, we isolated slow-growing NSLAB from cheeses collected in different regions of the United States, Australia, New Zealand, and Ireland. Seven of the cheeses showed significant gas defects and 12 did not. Nonstarter lactic acid bacteria were isolated from these cheeses on de Man, Rogosa, and Sharpe medium supplemented with ribose, a preferred substrate for Lb. wasatchensis. Identification was confirmed with 16S rRNA gene sequencing and the API50CH (bioMérieux, Marcy l'Etoile, France) carbohydrate panel. Isolates were also compared with one another by using repetitive element sequence-based PCR (rep-PCR). Lactobacillus wasatchensis was isolated only from cheeses demonstrating late-gas development and was found in samples from 6 of the 7 cheeses. This supports laboratory evidence that this organism is a causative agent of late gas production defects. The rep-PCR analysis produced distinct genetic fingerprints for isolates from each cheese, indicating that Lb. wasatchensis is found in several regions across the United States and is not a local phenomenon.

Off loading wheelchair cushion provides best case reduction in tissue deformation as indicated by MRI.

Off-loading or the Orthotic approach to wheelchair seating has been used successfully to provide seating that optimizes tissue protection at the ischial tuberosities (ITs), sacrum and greater trochanters. Recent publications indicate the significance of preventing tissue compression to reduce ulcer formation. Comparative Magnetic Resonance Imaging (MRI) of individuals seated on two cushion types provides direct evidence of tissue unloading resulting in the reduction in tissue compression. Measurement of tissue compression in MRI images provides the cumulative impact of compression and shear resulting in ultimate tissue thickness documented here. In this study's application of MRI to off-loading cushions (OLC), an alternate form of tissue protection was observed. Instead of incorporating immersion and envelopment, loads were transferred from high-risk areas, such as bony prominences, to lower risk soft tissues. This method shows both shearing and compression of load bearing tissues in seated individuals with the OLC in place. Tissue thickness measurements determined by MRI analysis indicate that the OLC provides greater reduction in tissue deformation than the air cell cushion (ACC). Deformation of tissues loaded by the OLC is not significantly different from the deformations seen with the ACC. This research represents the first reported use of MRI to document the comparative off-loading capabilities of two cushions and the resultant tissue compression and ulceration risk. While MRI analysis may not be incorporated in daily cushion prescription, this paper proposes a methodology in which MRI analysis of tissue deformation on comparative cushions allows the determination of best-case cushion selection for reduction of ischial pressure ulcer (PU) risk.

Lactobacillus wasatchensis sp. nov., a non-starter lactic acid bacteria isolated from aged Cheddar cheese.

A Gram-stain positive, rod-shaped, non-spore-forming strain (WDC04T), which may be associated with late gas production in cheese, was isolated from aged Cheddar cheese following incubation on MRS agar (pH 5.2) at 6 °C for 35 days. Strain WDC04T had 97 % 16S rRNA gene sequence similarity with Lactobacillus hokkaidonensis DSM 26202T, Lactobacillus oligofermentans 533, 'Lactobacillus danicus' 9M3, Lactobacillus suebicus CCUG 32233T and Lactobacillus vaccinostercus DSM 20634T. API 50 CH carbohydrate fermentation panels indicated strain WDC04T could only utilize one of the 50 substrates tested, ribose, although it does slowly utilize galactose. In the API ZYM system, strain WDC04T was positive for leucine arylamidase, valine arylamidase, cysteine arylamidase (weakly), naphthol-AS-BI-phosphohydrolase and ß-galactosidase activities. Total genomic DNA was sequenced from strain WDC04T using a whole-genome shotgun strategy on a 454 GS Titanium pyrosequencer. The sequence was assembled into a 1.90 Mbp draft genome consisting of 105 contigs with preliminary genome annotation performed using the RAST algorithm (rast.nmpdr.org). Genome analysis confirmed the pentose phosphate pathway for ribose metabolism as well as galactose, N-acetylglucosamine, and glycerol fermentation pathways. Genomic analysis places strain WDC04T in the obligately heterofermentative group of lactobacilli and metabolic results confirm this conclusion. The result of genome sequencing, along with 16S rRNA gene sequence analysis, indicates WDC04T represents a novel species of the genus Lactobacillus, for which the name Lactobacillus wasatchensis sp. nov. is proposed. The type strain is WDC04T ( = DSM 29958T = LMG 28678T).

Growth and gas formation by Lactobacillus wasatchensis, a novel obligatory heterofermentative nonstarter lactic acid bacterium, in Cheddar-style cheese made using a Streptococcus thermophilus starter.

A novel slow-growing, obligatory heterofermentative, nonstarter lactic acid bacterium (NSLAB), Lactobacillus wasatchensis WDC04, was studied for growth and gas production in Cheddar-style cheese made using Streptococcus thermophilus as the starter culture. Cheesemaking trials were conducted using S. thermophilus alone or in combination with Lb. wasatchensis deliberately added to cheese milk at a level of ~10(4) cfu/mL. Resulting cheeses were ripened at 6 or 12°C. At d 1, starter streptococcal numbers were similar in both cheeses (~10(9) cfu/g) and fast-growing NSLAB lactobacilli counts were below detectable levels (<10(2) cfu/g). As expected, Lactobacillus wasatchensis counts were 3×10(5) cfu/g in cheeses inoculated with this bacterium and below enumeration limits in the control cheese. Starter streptococci decreased over time at both storage temperatures but declined more rapidly at 12°C, especially in cheese also containing Lb. wasatchensis. Populations of fast-growing NSLAB and the slow-growing Lb. wasatchensis reached 5×10(7) and 2×10(8) cfu/g, respectively, after 16 wk of storage at 12°C. Growth of NSLAB coincided with a reduction in galactose concentration in the cheese from 0.6 to 0.1%. Levels of galactose at 6°C had similar decrease. Gas formation and textural defects were only observed in cheese with added Lb. wasatchensis ripened at 12°C. Use of S. thermophilus as starter culture resulted in galactose accumulation that Lb. wasatchensis can use to produce CO2, which contributes to late gas blowing in Cheddar-style cheeses, especially when the cheese is ripened at elevated temperature.

Late blowing of Cheddar cheese induced by accelerated ripening and ribose and galactose supplementation in presence of a novel obligatory heterofermentative nonstarter Lactobacillus wasatchensis.

Lactobacillus wasatchensis sp. nov. has been studied for growth and gas formation in a control Cheddar cheese and in cheese supplemented with 0.5% ribose, 0.5% galactose, or 0.25% ribose plus 0.25% galactose using regular and accelerated cheese ripening temperatures of 6 and 12°C, respectively. Milk was inoculated with (1) Lactococcus lactis starter culture, or (2) Lc. lactis starter culture plus Lb. wasatchensis (10(4) cfu/mL). In the control cheese with no added Lb. wasatchensis, starter numbers decreased from 10(7) initially to ~10(4) cfu/g over 23 wk of ripening at 6°C. When the cheese was ripened at 12°C, or if Lb. wasatchensis was added, the final starter counts were 1 log lower. In contrast, nonstarter lactic acid bacteria in the cheese increased from <10(2) cfu/g at press to 10(6) to 10(7) cfu/g after 23 wk, with higher numbers being observed with ripening at 12°C. In cheese with no added Lb. wasatchensis, levels of Lb. wasatchensis were initially below the enumeration threshold but counts of up to 10(3) cfu/g were detected after 23 wk. When the cheese was inoculated with Lb. wasatchensis, it could be enumerated throughout ripening, with final levels at 23 wk being dependent on whether ribose had been added to the cheese curd. With added ribose (with or without added galactose), Lb. wasatchensis grew to 10(7) to 10(8) cfu/g after 23 wk, whereas without added ribose it was 1 log lower. In all cheeses with added Lb. wasatchensis, greater gas formation was observed at 12°C, with most gas production occurring after ~16 wk. Very little gas production was detected in cheese without added Lb. wasatchensis ripened at 12°C or in cheese with added Lb. wasatchensis ripened at 6°C. Adding a combination of ribose and galactose caused more gas formation, putatively because of the ability of Lb. wasatchensis to co-utilize both sugars and grow to high numbers, and then produce gas from galactose as ribose levels were depleted. Even without sugar supplementation, gas was observed in cheese with added Lb. wasatchensis after 16 wk. We also observed that Lb. wasatchensis could grow to high cell densities when grown in carbohydrate-restricted broth containing lactococcal cell lysate. This suggests that during cheese ripening, lysis of starter bacteria provides sufficient substrates (such as ribose) to allow growth of Lb. wasatchensis and, if fermentable hexose is available, the cheese will become gassy. We conclude that Lb. wasatchensis is a previously undetected contributor to late gas formation in Cheddar cheese and the defect is more pronounced when elevated ripening temperatures are used.

Growth and gas production of a novel obligatory heterofermentative Cheddar cheese nonstarter lactobacilli species on ribose and galactose.

An obligatory heterofermentative lactic acid bacterium, Lactobacillus wasatchii sp. nov., isolated from gassy Cheddar cheese was studied for growth, gas formation, salt tolerance, and survival against pasteurization treatments at 63°C and 72°C. Initially, Lb. wasatchii was thought to use only ribose as a sugar source and we were interested in whether it could also utilize galactose. We conducted experiments to determine the rate and extent of growth and gas production in carbohydrate-restricted (CR) de Man, Rogosa, and Sharpe (MRS) medium under anaerobic conditions with various combinations of ribose and galactose at 12, 23, and 37°C, with 23°C being the optimum growth temperature of Lb. wasatchii among the 3 temperatures studied. When Lb. wasatchii was grown on ribose (0.1, 0.5, and 1%), maximum specific growth rates (µmax) within each temperature were similar. When galactose was the only sugar, compared with ribose, µmax was 2 to 4 times lower. At all temperatures, the highest final cell densities (optical density at 640 nm) of Lb. wasatchii were achieved in CR-MRS plus 1% ribose, 0.5% ribose and 0.5% galactose, or 1% ribose and 1% galactose. Similar µmax values and final cell densities were achieved when 50% of the ribose in CR-MRS was substituted with galactose. Such enhanced utilization of galactose in the presence of ribose to support bacterial growth has not previously been reported. It appears that Lb. wasatchii co-metabolizes ribose and galactose, utilizing ribose for energy and galactose for other functions such as cell wall biosynthesis. Co-utilization of both sugars could be an adaptation mechanism of Lb. wasatchii to the cheese environment to efficiently ferment available sugars for maximizing metabolism and growth. As expected, gas formation by the heterofermenter was observed only when galactose was present in the medium. Growth experiments with MRS plus 1.5% ribose at pH 5.2 or 6.5 with 0, 1, 2, 3, 4, or 5% NaCl revealed that Lb. wasatchii is able to grow under salt and pH conditions typical of Cheddar cheese (4 to 5% salt-in-moisture, pH ~5.2). Finally, we found that Lb. wasatchii cannot survive low-temperature, long-time pasteurization but survives high-temperature, short-time (HTST) laboratory pasteurization, under which a 4.5 log reduction occurred. The ability of Lb. wasatchii to survive HTST pasteurization and grow under cheese ripening conditions implies that the presence of this nonstarter lactic acid bacterium can be a serious contributor to gas formation and textural defects in Cheddar cheese.