Participants in the Trans-Antarctic Winter Traverse Expedition Showed Increased Bacterial Load and Diversity in Saliva but Maintained Individual Differences within Stool Microbiota and Across Metabolite Fingerprints.

Understanding the impact of long-term physiological and environmental stress on the human microbiota and metabolome may be important for the success of space flight. This work is logistically difficult and has a limited number of available participants. Terrestrial analogies present important opportunities to understand changes in the microbiota and metabolome and how this may impact participant health and fitness. Here, we present work from one such analogy: the Transarctic Winter Traverse expedition, which we believe is the first assessment of the microbiota and metabolome from different bodily locations during prolonged environmental and physiological stress. Bacterial load and diversity were significantly higher during the expedition when compared with baseline levels (p < 0.001) in saliva but not stool, and only a single operational taxonomic unit assigned to the Ruminococcaceae family shows significantly altered levels in stool (p < 0.001). Metabolite fingerprints show the maintenance of individual differences across saliva, stool, and plasma samples when analysed using flow infusion electrospray mass spectrometry and Fourier transform infrared spectroscopy. Significant activity-associated changes in terms of both bacterial diversity and load are seen in saliva but not in stool, and participant differences in metabolite fingerprints persist across all three sample types.

Effectiveness of pediatric rotary, rotary and reciprocating instrumentations on bacterial load reduction in primary molars: an ex vivo comparative study.

This study aimed to evaluate the efficacy of intracanal Enterococcus faecalis reduction using pediatric rotary (EndoArt Pedo Kit Blue, EasyInSmile X-Baby and Denco Kids), rotary (ProTaper Next) and reciprocating (WaveOne Gold) file systems through microbiological analyses in primary molars. Seventy-five mandibular primary second molars were selected and divided into five instrumentation groups and a negative control group. After incubation, five roots were used to confirm biofilm formation on the root canals. Before and after instrumentation, bacterial samples were collected. The bacterial load reduction was statistically analyzed by using Kruskall-Wallis and Dunn post hoc tests at a significance level of 0.05. Denco Kids and EndoArt Pedo Kit Blue promoted higher bacterial reduction than EasyInSmile X-Baby systems. There was no difference in bacterial reduction between ProTaper Next rotary file systems and other groups. Among the single-file techniques, instrumentation with the Denco Kids rotary system showed a more significant bacterial load reduction than WaveOne Gold (p < 0.05). All systems used in the study reduced bacterial counts from root canals in primary teeth. Further studies are required to generate more information about the use of pediatric rotary file systems in clinics.

Performance of Tuberculosis Molecular Bacterial Load Assay Compared to Alere TB-LAM in Urine of Pulmonary Tuberculosis Patients with HIV Co-Infections.

Alternative tools are needed to improve the detection of M. tuberculosis (M. tb) in HIV co-infections. We evaluated the utility of Tuberculosis Molecular Bacterial Load Assay (TB-MBLA) compared to lipoarabinomannan (LAM) to detect M. tb in urine. Sputum Xpert MTB/RIF-positive patients were consented to provide urine at baseline, weeks 2, 8, 16, and 24 of treatment for TB-MBLA, culture, and LAM. Results were compared with sputum cultures and microscopy. Initial M. tb. H37Rv spiking experiments were performed to validate the tests. A total of 63 urine samples from 47 patients were analyzed. The median age (IQR) was 38 (30-41) years; 25 (53.2%) were male, 3 (6.5%) had urine for all visits, 45 (95.7%) were HIV positive, of whom 18 (40%) had CD4 cell counts below 200 cells/µL, and 33 (73.3%) were on ART at enrollment. Overall urine LAM positivity was 14.3% compared to 4.8% with TB-MBLA. Culture and microscopy of their sputum counterparts were positive in 20.6% and 12.7% of patients, respectively. Of the three patients with urine and sputum at baseline, one (33.33%) had urine TB-MBLA and LAM positive compared to 100% with sputum MGIT culture positive. Spearman's rank correction coefficient (r) between TB-MBLA and MGIT was -0.85 and 0.89 with a solid culture, p > 0.05. TB-MBLA has the promising potential to improve M. tb detection in urine of HIV-co-infected patients and complement current TB diagnostics.

Efficacy of lyophilised bacteria-rich faecal sediment and supernatant with reduced bacterial count for treating patients with Clostridioides difficile Infection - A novel method for capsule faecal microbiota transfer.

Background and aims: Faecal microbiota transfer (FMT) has managed to earn its place in the Clostridioides difficile infection (CDI) guidelines by having comparable efficacy and recurrence rate of fidaxomicin. After more than 100 successful FMT administration through nasogastric tube, we started using hard gelatine capsules filled with lyophilised faecal sediment and supernatant. Our main question was whether uncoated capsules (containing faecal sediment or supernatant) are comparable to the widely used nasogastric tubes in CDI. We also investigated the effect of storage and time on the survival rate of bacteria in the samples. Methods: We compared the efficacy of our capsules to other treatment options of CDI at the Department of Infectology at the University of Pécs (Hungary). For our study, stool was collected from a single donor. We treated 10 patients with relapsing CDI, 5 of them received supernatant, 5 received sediment. Donor samples were stored on 4 different temperatures and tested to determine the survival rates of bacteria. As pilot projects, we also assessed the changes of bacterial taxa, protein- and lipid compositions. Moreover, we selected 4 patients to compare their samples prior and after FMT by using microbiome (16S amplicon sequencing), protein, and lipid analyses. Results: 4 out of the 5 patients who received supernatant became symptomless within 2 days after FMT. In the sediment group 3 out of 5 patients were cured from CDI. Comparing the supernatant to the sediment, we found significantly lower number of colony-forming units in the supernatant. We found that -80°C is the most suitable temperature to store the samples. The stool lipid profiles of recipients showed a more diverse composition after FMT, and changes in the stool protein profiles were observed as well. In the microbiome analysis, we observed an increase in the alpha diversity after FMT. Conclusions: Our study of 10 patients showed good efficacy of lyophilised faecal supernatant using capsules. The single donor approach proved to be effective in our investigation. A significantly lower CFU number was sufficient for the effect, the separation can be achieved by widely available instruments. For storage temperature, -20°C was sufficient in our clinical practice.

Chlamydia trachomatis Coinfection Does Not Influence Mycoplasma genitalium Bacterial Load in Urogenital Samples.

BACKGROUND: Mycoplasma genitalium (MG) is associated with urethritis in men and weakly associated with pelvic inflammatory disease in women. Mycoplasma genitalium coinfections with Neisseria gonorrhoeae (NG) and Chlamydia trachomatis (CT) are commonly reported; however, little is known about their interaction. One study suggested that MG/NG coinfections might increase the bacterial load of NG, which has been shown to have a higher transmission potential. As even less is known about the impact of a simultaneous MG/CT infection, we assessed whether patients with urogenital MG/CT coinfections have a higher bacterial load than patients with a single infection. METHODS: There were 1673 urogenital samples from patients from a population-based chlamydia study, and our sexually transmitted infection clinic tested for both CT and MG. When positive, the load was quantified. Nonparametric tests compared the CT and MG load, and linear regression analyses tested the association of the CT and MG load within a patient. RESULTS: In 60 MG-positive patients, MG load ranged from 1.7 to 6.0 log10 copies/ml, similar to the CT load distribution. Only 6 patients were MG-positive and CT-negative, but the MG load distribution was similar to that of CT-positive patients (n.s.). The MG and CT load was unrelated in coinfected persons (n.s.). CONCLUSIONS: We found no correlation between the CT and MG load in urogenital samples, and the MG load distribution was similar in CT-positive and CT-negative patients. These results could have implications for the transmission risk of these infections.

Sputum bacterial load and bacterial composition correlate with lung function and are altered by long-term azithromycin treatment in children with HIV-associated chronic lung disease.

BACKGROUND: Long-term azithromycin (AZM) treatment reduces the frequency of acute respiratory exacerbation in children and adolescents with HIV-associated chronic lung disease (HCLD). However, the impact of this treatment on the respiratory bacteriome is unknown. METHOD: African children with HCLD (defined as forced expiratory volume in 1 s z-score (FEV1z) less than - 1.0 with no reversibility) were enrolled in a placebo-controlled trial of once-weekly AZM given for 48-weeks (BREATHE trial). Sputum samples were collected at baseline, 48 weeks (end of treatment) and 72 weeks (6 months post-intervention in participants who reached this timepoint before trial conclusion). Sputum bacterial load and bacteriome profiles were determined using 16S rRNA gene qPCR and V4 region amplicon sequencing, respectively. The primary outcomes were within-participant and within-arm (AZM vs placebo) changes in the sputum bacteriome measured across baseline, 48 weeks and 72 weeks. Associations between clinical or socio-demographic factors and bacteriome profiles were also assessed using linear regression. RESULTS: In total, 347 participants (median age: 15.3 years, interquartile range [12.7-17.7]) were enrolled and randomised to AZM (173) or placebo (174). After 48 weeks, participants in the AZM arm had reduced sputum bacterial load vs placebo arm (16S rRNA copies/µl in log10, mean difference and 95% confidence interval [CI] of AZM vs placebo - 0.54 [- 0.71; - 0.36]). Shannon alpha diversity remained stable in the AZM arm but declined in the placebo arm between baseline and 48 weeks (3.03 vs. 2.80, p = 0.04, Wilcoxon paired test). Bacterial community structure changed in the AZM arm at 48 weeks compared with baseline (PERMANOVA test p = 0.003) but resolved at 72 weeks. The relative abundances of genera previously associated with HCLD decreased in the AZM arm at 48 weeks compared with baseline, including Haemophilus (17.9% vs. 25.8%, p < 0.05, ANCOM ω = 32) and Moraxella (1% vs. 1.9%, p < 0.05, ANCOM ω = 47). This reduction was sustained at 72 weeks relative to baseline. Lung function (FEV1z) was negatively associated with bacterial load (coefficient, [CI]: - 0.09 [- 0.16; - 0.02]) and positively associated with Shannon diversity (0.19 [0.12; 0.27]). The relative abundance of Neisseria (coefficient, [standard error]: (2.85, [0.7], q = 0.01), and Haemophilus (- 6.1, [1.2], q < 0.001) were positively and negatively associated with FEV1z, respectively. An increase in the relative abundance of Streptococcus from baseline to 48 weeks was associated with improvement in FEV1z (3.2 [1.11], q = 0.01) whilst an increase in Moraxella was associated with decline in FEV1z (-2.74 [0.74], q = 0.002). CONCLUSIONS: AZM treatment preserved sputum bacterial diversity and reduced the relative abundances of the HCLD-associated genera Haemophilus and Moraxella. These bacteriological effects were associated with improvement in lung function and may account for reduced respiratory exacerbations associated with AZM treatment of children with HCLD. Video Abstract.

[Profiles and differences of submucosal microbial in peri-implantitis and health implants: A cross-sectional study].

OBJECTIVE: To describe the submucosal microbial profiles of peri-implantitis and healthy implants, and to explore bacteria that might be correlated with clinical parameters. METHODS: In the present cross-sectional study, 49 patients were recruited. Each patient contributed with one implant, submucosal biofilms were collected from 20 healthy implants and 29 implants with peri-implantitis. DNA was extracted and bacterial 16S ribosomal RNA (16S rRNA) genes were amplified. Submucosal biofilms were analyzed using 16S rRNA sequencing at Illumina MiSeq platform. Differences between the groups were determined by analyzing α diversity, microbial component and microbial structure. The potential correlation between the bacteria with pocket probing depth (PPD) of peri-implant calculated by Spearman correlation analysis. RESULTS: The α diversity of submucosal microbial of health group was significantly lower than that in peri-implantitis group (Chao1 index: 236.85±66.13 vs. 150.54±57.43, P < 0.001; Shannon index: 3.42±0.48 vs. 3.02±0.65, P=0.032). Principal coordinated analysis showed that the submucosal microbial structure had significant difference between healthy and peri-implantitis groups [R2=0.243, P=0.001, analysis of similarities (ANOSIM)]. Compared with healthy implants, relative abundance of periodontal pathogens were higher in peri-implantitis, including members of the red complex (Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola) and some members of orange complex (Precotella intermedia, Eubacterium nodatum, Parvimonas micra), as well as some new periodontal pathogens, such as Fillifactor alocis, Fretibacterium fastidiosum, Desulfobulbus sp._HMT_041, and Porphyromonas endodontalis. Spearman correlation analysis revealed that the relative abundance of Treponema denticola (r=0.686, P < 0.001), Tannerella forsythia (r=0.675, P < 0.001), Fretibacterium sp. (r=0.671, P < 0.001), Desulfobulbus sp._HMT_041 (r=0.664, P < 0.001), Filifactor alocis (r=0.642, P < 0.001), Fretibacterium fastidiosum (r=0.604, P < 0.001), Porphyromonas gingivalis (r=0.597, P < 0.001), Porphyromonas endodontalis (r=0.573, P < 0.001) were positive correlated with PPD. While the relative abundance of Rothia aeria (r=-0.615, P < 0.001) showed negatively correlation with PPD. CONCLUSION: Marked differences were observed in the microbial profiles of healthy implants and peri-implantitis. The members of red and orange complex as well as some new periodontal pathogens seem to play an important role in peri-implant disease. Compared with healthy implants, the submucosal microbial of peri-implantitis were characterized by high species richness and diversity.

Effectiveness of a Phage Cocktail as a Potential Biocontrol Agent against Saprophytic Bacteria in Ready-To-Eat Plant-Based Food.

This study aimed to evaluate the effectiveness of the phage cocktail to improve the microbiological quality of five different mixed-leaf salads: rucola, mixed-leaf salad with carrot, mixed-leaf salad with beetroot, washed and unwashed spinach, during storage in refrigerated conditions. Enterobacterales rods constituted a significant group of bacteria in the tested products. Selected bacteria were tested for antibiotic resistance profiles and then used to search for specific bacteriophages. Forty-three phages targeting bacteria dominant in mixed-leaf salads were isolated from sewage. Their titer was determined, and lytic activity was assessed using the Bioscreen C Pro automated growth analyzer. Two methods of phage cocktail application including spraying, and an absorption pad were effective for rucola, mixed leaf salad with carrot, and mixed leaf salad with beetroot. The maximum reduction level after 48 h of incubation reached 99.9% compared to the control sample. In washed and unwashed spinach, attempts to reduce the number of microorganisms did not bring the desired effect. The decrease in bacteria count in the lettuce mixes depended on the composition of the autochthonous saprophytic bacteria species. Both phage cocktail application methods effectively improved the microbiological quality of minimally processed products. Whole-spectral phage cocktail application may constitute an alternative food microbiological quality improvement method without affecting food properties.

Point-of-care fluorescence imaging reveals extent of bacterial load in diabetic foot ulcers.

Elevated levels of bacteria, including biofilm, increase the risk of chronic wound infection and inhibit healing. Addressing asymptomatic high bacterial loads is challenged by a lack of clinical terminology and diagnostic tools. This post-hoc multicenter clinical trial analysis of 138 diabetic foot ulcers investigates fluorescence (FL)-imaging role in detecting biofilm-encased and planktonic bacteria in wounds at high loads. The sensitivity and specificity of clinical assessment and FL-imaging were compared across bacterial loads of concern (104 -109 CFU/g). Quantitative tissue culture confirmed the total loads. Bacterial presence was confirmed in 131/138 ulcers. Of these, 93.9% had loads >104 CFU/g. In those wounds, symptoms of infection were largely absent and did not correlate with, or increase proportionately with, bacterial loads at any threshold. FL-imaging increased sensitivity for the detection of bacteria across loads 104 -109 (P < .0001), peaking at 92.6% for >108 CFU/g. Imaging further showed that 84.2% of ulcers contained high loads in the periwound region. New terminology, chronic inhibitory bacterial load (CIBL), describes frequently asymptomatic, high bacterial loads in diabetic ulcers and periwound tissues, which require clinical intervention to prevent sequelae of infection. We anticipate this will spark a paradigm shift in assessment and management, enabling earlier intervention along the bacterial-infection continuum and supporting improved wound outcomes.

Agent-based model indicates chemoattractant signaling caused by Mycobacterium avium biofilms in the lung airway increases bacterial loads by spatially diverting macrophages.

Incidence and prevalence of MAC infections are increasing globally, and reinfection is common. Thus, MAC infections present a significant public health challenge. We quantify the impact of MAC biofilms and repeated exposure on infection progression using a computational model of MAC infection in lung airways. MAC biofilms aid epithelial cell invasion, cause premature macrophage apoptosis, and limit antibiotic efficacy. In this computational work we develop an agent-based model that incorporates the interactions between bacteria, biofilm, and immune cells. In this computational model, we perform virtual knockouts to quantify the effects of the biofilm sources (deposited with bacteria vs. formed in the airway), and their impacts on macrophages (inducing apoptosis and slowing phagocytosis). We also quantify the effects of repeated bacterial exposures to assess their impact on infection progression. Our simulations show that chemoattractants released by biofilm-induced apoptosis bias macrophage chemotaxis towards pockets of infected and apoptosed macrophages. This bias results in fewer macrophages finding extracellular bacteria, allowing the extracellular planktonic bacteria to replicate freely. These spatial macrophage trends are further exacerbated with repeated deposition of bacteria. Our model indicates that interventions to abrogate macrophages' apoptotic responses to bacterial biofilms and/or reduce frequency of patient exposure to bacteria will lower bacterial load, and likely overall risk of infection.

Invited review: Redefining raw milk quality-Evaluation of raw milk microbiological parameters to ensure high-quality processed dairy products.

Raw milk typically has little bacterial contamination as it leaves the udder of the animal; however, through a variety of pathways, it can become contaminated with bacteria originating from environmental sources, the cow herself, and contact with contaminated equipment. Although the types of bacteria found in raw milk are very diverse, select groups are particularly important from the perspective of finished product quality. In particular, psychrophilic and psychrotolerant bacteria that grow quickly at low temperatures (e.g., species in the genus Pseudomonas and the family Enterobacteriaceae) and produce heat-stable enzymes, and sporeforming bacteria that survive processing hurdles in spore form, are the 2 primary groups of bacteria related to effects on processed dairy products. Understanding factors leading to the presence of these important bacterial groups in raw milk is key to reducing their influence on processed dairy product quality. Here we examine the raw milk microbiological parameters used in the contemporary dairy industry for their utility in identifying raw milk supplies that will perform well in processed dairy products. We further recommend the use of a single microbiological indicator of raw milk quality, namely the total bacteria count, and call for the development of a whole-farm approach to raw milk quality that will use data-driven, risk-based tools integrated across the continuum from production to processing and shelf-life to ensure continuous improvement in dairy product quality.

Analysis of very low bacterial counts in small sample volumes using angle-resolved light scattering.

Because of its high sensitivity to even small objects and the quick measurement principle, angle-resolved scattering (ARS) measurements exhibit a promising potential as a rapid analysis tool for bacterial cells at small sample sizes and very low numbers of cells. In this study, investigations on scattered light from various bacterial cell samples revealed applicability down to single cell levels, which is a huge benefit compared to conventional methods that depend on time-consuming cellular growth over several hours or even days. With the proposed setup and data analysis method, it is possible to detect scatter differences among cell types, together with the cell concentration.

Comparison of Microorganism Detection, Time to Positivity, and Time-Dependent Shift in Viable Bacterial Count from VersaTREK and BacT/ALERT 3D Blood Culture Systems.

Rapid positive blood culture reporting allows early and appropriate treatment of severe infections to improve patient prognosis. This study evaluated performance of the VersaTREK system with gas pressure detection and tornado stirring method and the conventional BacT/ALERT 3D system. Time to positivity (TTP) of simulated blood cultures without whole blood using 17 ATCC strains was faster with VersaTREK than BacT/ALERT 3D, averaging 6.3 h in aerobic bottles and 12.7 hours in anaerobic bottles. In simulated blood cultures with whole blood using 53 clinical isolates, on average, VersaTREK was faster in aerobic bottles by 6.5 h but slower in anaerobic bottles by 3.8 h. Fifty of 53 simulated blood cultures with whole blood (94%) showed fastest TTP with VersaTREK. TTP of VersaTREK for anaerobic bacteria Bacteroides fragilis and Clostridium perfringens, Helicobacter cinaedi, and Candida glabrata was fast, and viable bacteria numbers in bottles using the Miles and Misra method increased quickly.

Immunization with a tri-antigen syphilis vaccine significantly attenuates chancre development, reduces bacterial load, and inhibits dissemination of Treponema pallidum.

Syphilis continues to be a significant public health concern worldwide. The disease is endemic in many low- and middle-income countries, and rates have risen sharply in high-income countries over the last decade. The continued prevalence of infectious and congenital syphilis worldwide highlights the need for the development of an effective syphilis vaccine to complement public health measures for syphilis control. The complex, multi-stage course of syphilis infection necessitates a holistic approach to the development of an effective vaccine, in which immunization prevents both the localized stage of infection (typified by the highly infectious chancre) and the disseminated stages of infection (typified by the secondary rash, neurosyphilis, and destructive tertiary lesions, as well as congenital syphilis). Inhibiting development of the infectious chancre would reduce transmission thus providing community- level protection, while preventing dissemination would provide individual-level protection by reducing serious sequelae and may also provide community level protection by reducing shedding during secondary syphilis. In the current study we build upon prior investigations which demonstrated that immunizations with individual, well characterized T. pallidum TprK, TprC, and Tp0751 peptides elicits partial protection against infection in the animal model. Specifically, we show here that immunization with a TprC/TprK/Tp0751 tri-antigen cocktail protects animals from progressive syphilis lesions and substantially inhibits dissemination of the infection.

Growth Prediction of the Total Bacterial Count in Freshly Squeezed Strawberry Juice during Cold Storage Using Electronic Nose and Electronic Tongue.

The growth models of total bacterial count in freshly squeezed strawberry juice were established by gas and taste sensors in this paper. By selecting the optimal sensors and fusing the response values, the Modified Gompertz, Logistic, Huang and Baranyi models were used to predict and simulate the growth of bacteria. The results showed that the R2 values for fitting the growth model of total bacterial count of the sensor S7 (an electronic nose sensor), of sweetness and of the principal components scores were 0.890-0.944, 0.861-0.885 and 0.954-0.964, respectively. The correlation coefficients, or R-values, between models fitted by the response values and total bacterial count ranged from 0.815 to 0.999. A single system of electronic nose (E-nose) or electronic tongue (E-tongue) sensors could be used to predict the total bacterial count in freshly squeezed strawberry juice during cold storage, while the higher rate was gained by the combination of these two systems. The fusion of E-nose and E-tongue had the best fitting-precision in predicting the total bacterial count in freshly squeezed strawberry juice during cold storage. This study proved that it was feasible to predict the growth of bacteria in freshly squeezed strawberry juice using E-nose and E-tongue sensors.

Efficacy of Cold Atmospheric Plasma Therapy on Chronic Wounds: An Updated Systematic Review and Meta-Analysis of RCTs.

Objective: A previous meta-analysis has revealed that cold atmospheric plasma (CAP) might not be clinically beneficial to chronic wounds. However, several new randomized controlled trials (RCTs) reported that CAP was an effective treatment option for accelerating wound healing in chronic wounds. The purpose of this review is to incorporate these new results and evaluate the efficacy of CAP in chronic wounds. Methods: The major databases, including PubMed, Embase, Cochrane Library, and Web of Science, were searched for articles related to CAP treatment in chronic wounds until March 21, 2022. The literature retrieval and evaluation were carried out by two independent researchers. Result: A total of 13 randomized clinical trials published between 2010 and 2022 were finally included. CAP therapy showed to be more effective in reducing the area of wounds (mean difference (MD): -1.74, 95%; confidence interval (CI): [-3.14, -0.33], p = 0.02), compared with non-CAP treatments. The immediate reduction of the bacterial load was higher in the CAP group than in the control group. (MD: -0.37, 95%; CI: [-0.7, -0.05], p = 0.02). Conclusion: No significant changes were found in long-term antibacterial efficacy and pain perception between the two groups. However, more RCTs of excellent methodological quality are required to confirm technical details of the source of AP and the appropriate duration of the treatment with plasma.

Disinfection of needleless connectors to reduce Staphylococcus aureus bacterial load.

HIGHLIGHTS: Compare effectiveness of chemical disinfectants in reducing S. aureus. Five disinfectants reduced the bacterial load, especially chlorhexidine solutions. Focus on Brazilian clinical practice of needleless connector disinfection. PURPOSE: This study aimed to gain further knowledge about the comparative effectiveness of chemical disinfectants in reducing the bacterial load of NCs inoculated with S. aureus. METHODS: Disinfection of needleless connectors was undertaken in vitro against S. aureus comparing 70% isopropyl alcohol (IPA), 70% ethanol, 0.5% and 2% chlorhexidine in 70% IPA applied with gauze, and 70% IPA single-use cap (Site-Scrub®). RESULTS: All disinfectants reduced the bacterial load (P<0.001), especially the chlorhexidine solutions. Mechanical friction should follow guidelines. CONCLUSION: This study found that all tested disinfectants effectively reduced the bacterial load and more clinical studies must be developed with a focus on the Brazilian clinical practice of needleless connector disinfection.

Bacterial Milk Quality along the Value Chain in Smallholder Dairy Production.

This study aimed to evaluate the microbial quality of raw milk along the milk value chain at Africa University (AU). Eighteen Holstein-Friesian cows were used in this experiment. A total of 270 milk samples were collected for laboratory analysis at three different stages, during milking (DM), from the bulk tank (BT), and at the dining hall (DH), to determine the total bacterial count (TBC), Escherichia coli, and Salmonella enterica. Samples were cultured in Petri dishes using an appropriate medium for 48 hours. The plate count method was used to determine the quantity of bacteria. Data were analyzed using GLM SPSS. The results indicated that TBC increased (P < 0.05) from one site to the next along the value chain, yet it undulates when measured over time. Escherichia. coli and S. enterica counts increased (P < 0.05) at the last site of collection and the highest counts were recorded in week two. In conclusion, the current study indicates the hygiene of the dairy parlor with very low TBC, E. coli, and S. enterica counts during milking and bulk tank storage and that the relationship between TBC and E. coli is nonlinear with respect to time.

A novel approach to screening and managing the urinary tract infections suspected sample in the general human population.

Background: Automated urine technology providing standard urinalysis data can be used to support clinicians in screening and managing a UTI-suspected sample. Fully automated urinalysis systems have expanded in laboratory practice. Commonly used were devices based on digital imaging with automatic particle recognition, which expresses urinary sediment results on an ordinal scale. There were introduced fluorescent flow cytometry analyzers reporting all parameters quantitatively. There is a need to harmonize the result and support comparing bacteria and WBC qualitative versus semiquantitative results. Methods: A total of 1,131 urine samples were analyzed on both automated urinalysis systems. The chemical components of urinalysis (leukocyte esterase and nitrate reductase) and the sediment results (leukocytes and bacteria) were investigated as potential UTI indicators. Additionally, 106 specimens were analyzed on UF-5000 and compared with culture plating to establish cut-offs that can be suitable for standard urinalysis requirements and help to guide on how to interpret urinalysis results in the context of cultivation reflex. Results: The medians of bacteria counts varies from 16.2 (absence), 43.0 (trace), 443.5 (few), 5,389.2 (moderate), 19,356.6 (many) to 32,545.2 (massive) for particular digital microscopic bacteriuria thresholds. For pyuria thresholds, the medians of WBC counts varies from 0.8 (absence), 2.0 (0-1), 7.7 (2-3), 21.3 (4-6), 38.9 (7-10), 61.3 (11-15) to 242.2 (>30). Comparing the culture and FFC data (bacterial and/or WBC counts) was performed. Satisfactory sensitivity (100%), specificity (83.7%), negative predictive value (100%), and positive predictive value (75%) were obtained using indicators with the following cut-off values: leukocytes ≥40/µl or bacteria ≥300/µl. Conclusions: Accurate urinalysis gives information about the count of bacteria and leukocytes as useful indicators in UTIs, in general practice it can be a future tool to cross-link clinical and microbiology laboratories. However, the cut-off adjustments require individual optimization.

The microbial condition of Scottish wild deer carcasses collected for human consumption and the hygiene risk factors associated with Escherichia coli and total coliforms contamination.

Wild deer hunting is necessary in Scotland to control deer population density, with most carcasses being processed for human consumption. As limited information is available on the microbial condition of Scottish venison, we studied the variation of total coliforms and Escherichia coli (E. coli) on 214 wild deer carcasses collected from six approved establishments. Samples were collected from the hide, body cavity and external surface of each carcass and mean values were determined following bacterial plate counts. The mean log10/cm2 coliforms were 5.78 (hide), 6.80 (body cavity) and 6.36 (external surface). The mean log10/cm2E. coli were 1.82 (hide), 2.27 (body cavity) and 2.17 (external carcass). Significantly higher coliforms counts were associated with storage-to-dressing times above 6 days and with longer transport distances. Risk factors that increased E. coli were red deer species, ambient temperature above 7 °C during hunting, dirty hides, faecal contamination and moisture or slimy film on the carcass. Although the bacterial counts obtained in this study indicated some hygienic processing, for around half of the carcasses, the E. coli counts were above 2 log10/cm2. Therefore, the above risk factors suggest a few handling hygiene practices that should be further improved to enhance quality and safety.