Presence Of Pathogenic Bacteria Research Articles

Study on Presence of Pathogenic Bacteria in Serving Cutleries from Food Outlets in and Around a Medical College

Study on Presence of Pathogenic Bacteria in Serving Cutleries from Food Outlets in and Around a Medical College

QUALIDADE HIGIÊNICO-SANITÁRIA DE TAMBAQUI (Colossoma macropomum) COMERCIALIZADO NA CIDADE DE SÃO LUÍS - MA

Resumo A identificação das condições higiênico-sanitárias do pescado é de suma importância para a promoção da saúde e a qualidade de vida da população. Dessa forma, o presente trabalho teve como objetivo avaliar a qualidade microbiológica e verificar o perfil de susceptibilidade a antimicrobianos das cepas isoladas de amostras de tambaqui (C. macropomum) comercializadas na cidade de São Luís, Maranhão. As 60 amostras de tambaqui, obtidas de supermercados e feiras livres, foram analisadas quanto à determinação do Número Mais Provável (NMP) de coliformes totais e termotolerantes; pesquisa de Escherichia coli; pesquisa de Aeromonas spp., e perfil de resistência antimicrobiana. Foi detectada uma elevada contagem de coliformes totais e termotolerantes, além de bactérias patogênicas, tais como: Aeromonas spp. em 56 (93,3%) e E. coli, em 13 (43,3%) das amostras analisadas. Em relação ao perfil de resistência antimicrobiana, os isolados de E. coli demonstraram baixos percentuais de resistência e as cepas de Aeromonas apresentaram elevados níveis de resistência aos antimicrobianos testados. Pode-se concluir que o tambaqui comercializado em São Luís − MA apresenta condições higiênico-sanitárias inadequadas para o consumo.

Assessment of Microbiological Quality of Air Environment Around Waste Dumpsites Within Keffi Metropolis in Northern Nigeria

This study is to assess the microbiological quality and public health hazard associated with the air environment around waste dumpsites in Keffi, metropolis Nigeria was conducted. A total of 25 samples were collected from five different waste dumpsites around five different locations within the metropolis (Angwan Lambu, Keffi Main market, University Main campus and Nasarawa Road). Preliminary cultural, morphological, biochemical characteristics of the isolates revealed the following isolates, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Proteus vulgaris, Bacillus spp, Micrococcus spp., Aspergillus niger, Alternaria spp., Penicillum spp., Mucor spp., Fusarium spp., Rhizopus stolonifer and Candida spp. The total bacterial counts from the air environments of these waste dumpsites ranged between 2.2-7.4 x 101 cfu/m3, while the total fungal counts ranged between 1.4-4.4 x 101 cfu/m3. The susceptibility of the bacterial isolates against commonly used antibiotics tested showed varying degree of susceptibility and it is in the following decreasing order; Proteus vulgaris (93.3%) > Staphylococcus aureus (55.6%) > Micrococcus spp. (43.3%) > Pseudomonas aeruginosa (42.8%) > Bacillus spp. (41.2%) > Escherichia coli (30.8%). Health risk assessment indicates the presence of pathogenic bacteria and moulds with serious health risk to human within the environment due to the possibility of cross contamination of foods sold in the open. Consequently, there is the need to periodically monitor the number of microorganisms within the air of these environments using the established threshold stipulated for microbial concentrations in air.

Health from the Hive: 5% Propolis Mouth Wash as an Adjunct in the Treatment of Chronic Generalized Gingivitis-A Randomized Controlled Clinical Trial

Background: Gingival diseases affect 80% of the adult population in India and are considered to be plaque initiated inflammatory conditions with the presence of pathogenic bacteria. The term, Gingivitis means inflammation of the gums or gingiva. Gingivitis is a non-destructive type of periodontal disease if left untreated gingivitis can progress to periodontitis. Thus it can eventually lead to loss of teeth. It is a well-known fact that all periodontitis are initiated by gingivitis. Objectives: To assess the effectiveness of 5% propolis mouth wash in chronic generalized gingivitis and also to evaluate and compare the effectiveness of 5% propolis mouth wash with respect to the chlorhexidine mouth wash. Method: A total of 45 randomly selected patients (lottery method) between the age group of 18-70 years were selected for the study and grouped as follows: Group I with 15 patients who were treated with 5% propolis mouth wash. Group II with 15 patients were treated with chlorhexidine mouth wash controlled group and Group III with 15 patients were treated with normal saline (placebo). Result: The results indicated that there was a significant improvement in clinical parameters (p<0.05) in group 1 at the end of the study. Conclusion: Our study data showed that propolis mouthwash is more effective than other mouth washes on plaque accumulation and gingival inflammation. The study suggests that propolis can be used as a natural mouthwash, an alternative to chemical mouthwashes, e.g., chlorhexidine.

PATHOGENS FROM DRINKING WATER

… Objectives: Main objective was to check drinking water for pathogenic bacterialload, their resistance to antimicrobials and to create awareness among the people of Quetta cityabout safe drinking water. Place and Duration of Study: The study was conducted inDepartment of Microbiology Bolan Medical Complex Hospital Quetta during the hot season inQuetta City (June- September 2013). Methodology: One hundred and twenty five (125) tapewater samples were collected aseptically in 200 ml sterile caped glass bottles from differentlocalities of Quetta city. Samples were passed through mille pore assembly containing 0.45 μmpore-sizecellulose nitrate sterile membrane filter (MF).Viable count technique was used forenumeration of water samples having high bacterial burden. Serological tests and analyticalprofile index API-20E (Biomerieux France) were used to identify pathogens according to themanufacturer’s directions. Standardized antibiotic sensitivity test was performed on MuellerHinton agar using disc diffusion Kirby Bauer technique and McFarland Turbidity Standardmethod 0.5 following CLSI protocols. Results: Out of hundred and twenty five (125) tape watersamples 110 (88 %) showed highly pathogenic bacterial load, in which the most prominentorganism was E.coli 36 (28.8 %), followed by Enterobacter 35 (28 %), Klebsiella 24 (19.2 %),Pseudomonas, 10 (08 %), and Salmonella 05 (04 %). All pathogens in this study expressed a highlevel of resistance to antimicrobials that are commonly used in clinical medicine i.e. Tetracycline,Gentamycin, Sulphamethaxazole, Piperacillin, Ampicillin, Augmentin and Imipenam etc. Only 15(12 %) samples were pathogens free. Conclusion: Among drinking water samples the presenceof pathogenic bacteria (88%) is alarming for public health authorities. The emergence ofresistance and decreasing level of susceptibility of pathogens to a wide spectrum ofantimicrobials is a matter of great concern, because it may limit the availability of antimicrobialsfor clinical management of water born outbreaks in future.

Microbial Contamination of Organically and Conventionally Produced Fresh Vegetable Salads and Herbs from Retail Markets in Southwest Germany.

A total of 189 samples of fresh products (leafy salads, ready-to-eat mixed salads, and fresh herbs) bought in retail in Southwest Germany were investigated for their microbiological quality and the presence of pathogenic bacteria, including Salmonella spp., Listeria monocytogenes, and presumptive Bacillus cereus. Total aerobic mesophilic plate counts (TAC) ranged from 5.5 to 9.6 log colony-forming units (CFUs) per gram. Enterobacteria and pseudomonads were the predominant microorganisms and were detected in all samples with counts between 5.0 and 9.2 log CFU/g. Strains of Escherichia coli were detected in 9 salad (7.9%) and 25 herb samples (33.3%). Significant differences in bacterial counts were found between conventionally and organically-grown products: in herbs the counts of moulds were significantly higher in organically-grown products, while E. coli was only detected in conventionally-grown products. In conventionally-grown salad samples, yeast counts were significantly higher. Salmonella Enteritidis was only detected in two conventionally- and in one organically-produced salad samples (2.6%). No coagulase-positive staphylococci were detected in fresh salads as well as in herbs. High levels of B. cereus sensu lato (≥3 log CFU/g) were detected in 19 vegetable salads (16.7%) and even in 55 samples of fresh herbs (73.3%). Listeria monocytogenes could not be detected in fresh herbs; however, three L. monocytogenes strains were isolated from two conventionally-produced salad samples and belonged to PCR serogroup IIa. Although our results indicate a high microbial load in fresh salads and herbs in Southwest Germany in 2015, the incidences of human pathogenic bacteria, that is, L. monocytogenes, Salmonella spp., and coagulase-positive staphylococci strains, were low.

Bacterial Analysis of Different Types of Milk (Pasteurized, Unpasteurized and Raw Milk) Consumed in Kathmandu Valley

Objectives: The presence of pathogenic bacteria in milk is the major public health concern resulting in food borne illness. The aim of this study is to determine the microbial quality of three different types of milk consumed in Kathmandu Valley with respect to the acceptable standard guideline and measure the antibiotic susceptibility pattern of the Escherichia coli and Staphylococcus aureus isolates.Methods: A total of 66 samples (16 pasteurized, 25 unpasteurized and 25 raw milk) were collected from various sites of Kathmandu Valley. Those samples were subjected for total plate count and total coliform count by pour plate method. Furthermore, identification was made for the presence of E. coli and S. aureus with biochemical tests.Results: The mean total plate count (TPC) of pasteurized, unpasteurized and raw milk was 1.2X106 cfu/ml, 2.3 X 107 cfu/ml and 2.0 X 107 cfu/ml respectively. And, the mean total coliform count (TCC) of pasteurized, unpasteurized and raw milk was 2.9 X 104cfu/ml, 6.3 X 105 cfu/ml and 1.6 X 105 cfu/ ml respectively. Coliforms were detected in 50%, 84% and 56% of the pasteurized, unpasteurized and raw milk sample respectively. E. coli and S. aureus were isolated from 18.8% and 12.5% of pasteurized, 40% and 16% of unpasteurized and 20% and 24% of the raw milk samples respectively. Among total E. coli isolates (n=18), 16.7% were susceptible to ampicillin whereas 100% isolates were susceptible to other tested antibiotics. Similarly, 33.3% and 66.7% of the isolated S. aureus were susceptible to penicillin and cefoxitin respectively, whereas all S. aureus isolates were sensitive to all other antibiotics.Conclusion: The mean value of TPC and TCC of pasteurized and raw milk exceed the standard guideline by FDA. Higher total plate count and presence of coliforms (also E. coli) and S. aureus in this study necessitates the close monitoring of the pasteurization process and post pasteurization process (packaging, transportation, storage etc.).

Bacteria that affects coral health with an emphasis on the Gulf of Mexico and the Caribbean Sea

Reefs are the most biologically diverse marine ecosystems; therefore, to conserve the vast array of marine species that depend on them is vital. Many coral reefs around the world are under threat of extinction due to factors related to climate change and anthropogenic action. Studies have shown that specific infections develop at normal ocean temperatures and that warmer temperatures make the disease even more virulent. Also, as temperature increases, the infection in already infected corals becomes lethal. Thermal stress and the presence of pathogenic bacteria from the contribution of organic matter generates pathological affections in coral tissue, such as the White Band, Black Band, Yellow Band, White Plague, and Bacterial Whitening. This stimulates, in the short term, loss of coral cover and in the long term, negative impacts on the faunal composition of marine ecosystems. This review aims to present scientific information generated in the identification of different pathogenic bacteria that affect the health of corals.

Detection and characterization of Lactobacillus spp. in the porcine seminal plasma and their influence on boar semen quality.

The presence of pathogenic bacteria in ejaculates has been a topic in boar semen preservation over the last decades. Since little information is available on commensal bacteria in boar semen, the aim of the present study was to identify commensal lactobacilli in fresh cryopreserved boar semen and to examine their influence on boar semen quality. Therefore, 111 boar ejaculates were investigated for the presence of Lactobacillus species. Thirty samples (27%) contained viable Lactobacillus species (e.g. L. amylovorus, L. animalis, L. reuteri and Weisella minor). L. animalis and L. buchneri DSM 32407 (isolated from the bovine uterus) qualified for further examinations based on their growth rate in six antibiotic-free boar semen extenders. After a 120 min short-term incubation with an antibiotic-free BTS-extender, progressive motility was diminished (P = 0.001) upon addition of 105 and 106 colony forming units (CFU/mL) L. animalis. The supplementation with L. buchneri DSM 32407 had no significant (P > 0.05) influence on sperm quality during short-term co-incubation. After 168 h long-term co-incubation, motility analysis revealed a negative (P = 0.026) impact of 105 CFU/mL L. buchneri DSM 32407. A concentration- and storage-dependent effect is particularly obvious (P < 0.001) using 106 CFU/mL L. buchneri DSM 32407. Most notably, the thermo-resistance (TRT) for 106 CFU/mL L. buchneri DSM 32407 (P = 0.001) was inferior to BTS with and without gentamicin after 72 and 168 h of semen co-incubation. The supplementation of 105 CFU/mL L. buchneri DSM 32407 impaired progressive motility to a lesser extent. The percentage of mitochondrially active spermatozoa after 96 h (P = 0.009) and membrane-intact spermatozoa after 168 h (P < 0.001) was lower when 106 CFU/mL L. buchneri DSM 32407 were suspended compared with all other groups. Finally, the addition of L. buchneri DSM 32407 to BTS-extended boar semen had no competitive effect on the total amount of bacteria 48 h after co-incubation. In summary, the present study demonstrated that there are Lactobacillus species present in the porcine seminal plasma, which can be cultivated using standard procedures. However, long-term co-incubation of lactic acid bacteria with spermatozoa had a negative influence on spermatozoa.

Impact of in vitro inoculation and dietary supplementation with Bacillus subtilis on sperm quality of aged White Leghorn roosters

SUMMARY The probiotic Bacillus subtilis improves broiler performance; however, its effects on rooster reproduction are unknown. Therefore, 2 experiments (EXP) were conducted to evaluate impacts of B. subtilis on poultry semen quality. In EXP 1, B. subtilis was cultured for 48 h to a concentration of 108 CFU/mL. Semen from 72-week-old White Leghorn roosters was pooled and diluted 10-fold with the following treatments: 1) saline control, 2) sterile broth, 3) culture of B. subtilis, 4) supernatant from the culture, and 5) bacterial pellet from the culture. Semen pH and the sperm quality index (SQI) were obtained at 0 and 10 min post dilution to analyze effects of exposure length. The entire experiment was replicated 3 times. Semen pH and SQI were not affected by the B. subtilis pellet as compared to saline control. However, pH and SQI for every treatment containing broth were lower than the saline or pellet treatments. For EXP 2, 42 individually caged White Leghorn roosters, 74 wk old, were fed either 0 or 4.5 × 104 CFU of B. subtilis/g of feed. For each of 4 wk, individual semen samples were analyzed for pH, semen volume, sperm concentration, sperm viability, and the SQI. Additionally, semen concentrations of Na+, Ca2+, K+, Cl−, CO2, and O2 were measured. In the last week, ejaculates were serially diluted and plated to determine Bacillus spp. counts. The dietary addition of B. subtilis did not alter sperm quality characteristics, seminal ion concentrations, or Bacillus spp. counts. In conclusion, neither direct exposure of sperm nor dietary exposure of roosters to B. subtilis alters sperm quality, possibly because this bacterium is indigenous to the rooster's reproductive tract and semen. Therefore, feeding B. subtilis to roosters may not negatively affect fertility and may be an acceptable method to decrease pathogens, because B. subtilis alters intestinal microbiota. Future studies should investigate the effect of this probiotic on semen microbiota, especially regarding the presence of pathogenic bacteria that threaten public health.

Correlation between crestal alveolar bone loss with intracanal bacteria and apical lesion area in necrotic teeth

ObjectiveThis study aimed to analyze the correlation between crestal alveolar bone loss with the presence of some bacterial species in root canals and the apical lesion area of necrotic teeth. DesignData from 20 patients with diagnosis of pulp necrosis and acute apical abscesses, without active periodontal diseases, were evaluated. Patients with history of antibiotic usage three months prior to the study, with exposed pulp cavity, and with probing depth >3 mm were not included. The root size, the distance between the bone crest to the tooth apex in the mesial and distal surfaces, and the apical lesion area were measured from standard periapical radiographies by a calibrated examiner. Root canal samples were collected using sterilized paper points. In multirooted teeth, the largest root canal was sampled. Culture, microbial isolation and identification by phenotypic methods were performed. Spearman correlation and exact Fischer test were calculated between higher/lower existing bone crests, according to the median and the presence of specific bacteria. ResultsNo statistically significant differences were found between occurrence of pathogenic bacteria, such as Porphyromonas gingivalis, Porphyromonas endodontalis, and Prevotella intermedia, and groups with higher/lower degree of bone loss (p > 0.05). A negative significant correlation was found between Parvimonas micra and periodontal bone loss (p = 0.02). Additionally, no statistically significant association was found between crestal bone loss and the apical lesion area. ConclusionsIt was concluded that, in patients without active periodontitis, the presence of pathogenic bacteria in the root canal was not correlated with periodontal bone loss.

Persistencia, internalización y translocación de Escherichia coli O157:H7, O157:H16 y O105ab en plantas y frutos de tomate (Solanum lycopersicum L.)

The presence of pathogenic bacteria, such as Escherichia coli affects the quality and safety of vegetables that are consumed fresh and is associated with serious health problems. The objective of this study was to determine if three different strains of E. coli can penetrate and remain in plants and tomato fruits. A completely randomized experimental design was followed for which a tomato crop (“Cid” variety) was established under greenhouse conditions and three treatments were evaluated, T1 (E. coli O157:H7), T2 (E. coli from tomato cultivation [EcT] O157:H16), T3 (E. coli from spinach cultivation [EcH] O105ab) and a T4 control, with 100 plants each and four forms of inoculation: in the substrate, steam, petiole and the peduncle. Samples were carried out in vegetative stage, flowering, fruiting and physiological maturity to quantify in petri dish CFU/g and know if the bacteria managed to move around and recover in root, stem, flower and fruit. The phylogenetic groups that corresponded to the bacteria recovered were confirmed by biochemical tests, serotyping and PCR. At 120 days the recovery of bacteria in the plant was 23% (E. coli O157:H7), 28% (EcT O157:H16) and 55% (EcH O105ab) whit inoculation to the substrate while the inoculation by puncture the recovery was (in the same order) of 5%, 3%, and 4% at 30 days; 37%, 35% and 30% at 90 days; and 42%, 39% and 13% at 65 days. The strains submit the ability to enter the tomato plant and to stay in it and transported to the fruit, without producing that indicate their presence.

Assessing the health status of farmed mussels (Mytilus galloprovincialis) through histological, microbiological and biomarker analyses.

The Gulf of La Spezia (northern Tyrrhenian Sea, Italy) is a commercially important area both as a shipping port and for mussel farming. Recently, there has been increased concern over environmental disturbances caused by anthropogenic activities such as ship traffic and dredging and the effects they have on the health of farmed mussels. This paper reports the results of microbiological and histological analyses, as well as of measurement of several biomarkers which were performed to assess the health status of mussels (Mytilus galloprovincialis) from four rearing sites in the Gulf of La Spezia. Mussels were collected between October 2015 and September 2016 and histological analyses (including gonadal maturation stage), as well as the presence of pathogenic bacteria (Vibrio splendidus clade, V. aestuarianus and V. harveyi), viruses (Herpes virus and ostreid Herpes virus 1) and protozoa (Marteilia spp., in the summer season only) were carried out on a monthly basis. Conversely, biomarker responses in haemocyte/haemolymph (total haemocyte count, haemocyte diameter and volume, lysozyme and lactate dehydrogenase activities in cell-free haemolymph, and micronuclei frequency) and in gills and digestive gland (cortisol-like steroids and lipid peroxidation levels), were evaluated bimonthly. Microbiological data indicated that mussels contain a reservoir of potentially pathogenic bacteria, viruses and protozoa that in certain environmental conditions may cause a weakening of the immune system of animals leading to mortality episodes. The percentage of parasites detected in the mussels was generally low (9.6% for Steinhausia mytilovum, that is 17 samples out of 177 examined females; 3.4% for Proctoeces maculatus; 0.9% for Mytilicola intestinalis and 2% for ciliated protozoa), while symbiont loads were higher (31% for Eugymnanthea inquilina and Urastoma cyprinae). Interestingly, a previously undescribed haplosporidian was detected in a single mussel sample (0.2%) and was confirmed by in situ hybridization. Cells morphologically similar to Perkinsus sp. trophozoites were observed in 0.7% of the mussels analysed; however, infection with Perkinsus spp. could neither be confirmed by ISH nor by PCR. Different pathological aspects, such as host defence responses and regressive/progressive changes were detected in the gills, digestive glands, gonads and mantle. Only one single case of disseminated neoplasia (0.2%) was observed. As for the biomarker evaluation, the MANOVA analysis revealed the statistically significant effect that the variable "sampling site" had on the biological parameter measured, thus suggesting that the multibiomarker approach was able to differentiate the rearing sites.

To Quest Common Dangerous Bacterial Attack in Drinking Water

Buried ground water is very indispensable because it is major sources for the uses of humans, animals, industry and domestic purposes. The presence of pathogenic bacteria in drinking water is one of the great issues now-adays. The present article summarizes the dangerous effects, common methods of detection, recent outbreaks and possible plan to control the contamination of various common bacteria in drinking water to alert general public to be safe and healthy.

Comparision of the Microbial Count in Supragingival Plaque, Gingival Crevicular Blood and Saliva Samples Immediately after Diode Laser (970 ± 15 nm) Application in Chronic Periodontitis Patients: A Randomized Controlled Split Mouth Clinical Trial

Background: Periodontal disease affects 80% of the adult population in the United States. Periodontal disease are biofilm initiated inflammatory conditions with presence of pathogenic bacteria. Studies shown that laser decontamination has effect on the bacteria within the sulcus, reducing the risk of bacteremia caused from instrumentation, and to lower the microcount in aerosols created during ultrasonic instrumentation.Aim of the study: To evaluate immediate effects of a diode laser (970 ± 15 nm) on the microbial count in Supra gingival plaque, crevicular blood and saliva samples of patients suffering from chronic periodontitis.Materials and methods: A total of 15 subjects were recruited for the study. Each patient’s mouth was divided equally into two halves which were allotted randomly to a group based on coin toss method. Quadrants in group I (test group) were subjected to diode laser debridement while those of group II (control group) were subjected to saline irrigation. Supra gingival plaque, saliva and crevicular blood samples were collected before and immediately after laser debridement and saline irrigation and subsequently subjected to microbial analysis.Results: Clinical observations showed a significant reduction in microbial count i.e. reduction in mean CFUs(CFU/ ml) in both the groups while statistically-significant reduction is seen in the test group. Test group showed significant reduction in both Supra gingival plaque samples and crevicular blood in comparison to that of saliva samples.Conclusion: Present study concludes that diode laser (970 ± 15 nm) application has immediate effects in reducing the microbial load in supra gingival plaque, crevicular blood and saliva samples in patients with chronic periodontitis compared to control group.

Presence of pathogenic bacteria in ice cubes and evaluation of their survival in different systems

In this study, 60 samples of ice cubes produced at different levels (domestic, restaurant and industrial facilities), within a restricted geographical area, were investigated for their general microbiological characteristics through the analysis of populations other than enteric bacteria. Total mesophilic bacteria were in the range 1.01 × 102–9.55 × 103, 3.12 × 102–6.31 × 103 and 1.30 × 102–3.99 × 103 CFU/100 mL of thawed ice from domestic freezer (DF), stock boxes (SB) for self-production performed with ice machines in bars and pubs, and from sales packages (SP) of industrial productions, respectively. Some DF and SP samples were negative for the presence of total psychrotrophic bacteria, showing that there are no specific microbial groups associated with ice. Pseudomonads were found in the majority of ice samples analyzed. The levels of contamination of the ice samples were significantly different among the three ice cube production levels. The samples produced at domestic level and those collected from bars and pubs were characterised by the highest cell densities. The colonies representative for the different bacterial morphologies were randomly picked up from plates, purified to homogeneity and subjected to the phenotypic and genotypic characterisation. Fifty-two strains representing 31 species of eight bacterial genera were identified, with the most numerous groups included in Pseudomonas, Staphylococcus, Bacillus and Acinetobacter. A consistent percentage of the microorganisms identified from ice are known agents of human infections, and their presence indicate an environmental contamination. In order to evaluate the effectiveness of the ice cubes to transfer pathogenic agents to consumers, a bar consumption was simulated with different drink systems added with ice cubes artificially contaminated with the strains found at dominant levels (Acinetobacter lwoffii ICE100, Bacillus cereus ICE170, Pseudomonas putida ICE224 and Staphylococcus haemolyticus ICE182), and the results showed a consistent reduction of bacterial risk due to alcohol, CO2, pH and antibacterial ingredients of vodka, whisky, Martini, peach tea, tonic water and coke.

Antimicrobial peptides secreted by equine mesenchymal stromal cells inhibit the growth of bacteria commonly found in skin wounds

BackgroundThe prevalence of chronic skin wounds in humans is high, and treatment is often complicated by the presence of pathogenic bacteria. Therefore, safe and innovative treatments to reduce the bacterial load in cutaneous wounds are needed. Mesenchymal stromal cells (MSC) are known to provide paracrine signals that act on resident skin cells to promote wound healing, but their potential antibacterial activities are not well described. The present study was designed to examine the antibacterial properties of MSC from horses, as this animal model offers a readily translatable model for MSC therapies in humans. Specifically, we aimed to (i) evaluate the in vitro effects of equine MSC on the growth of representative gram-negative and gram-positive bacterial species commonly found in skin wounds and (ii) define the mechanisms by which MSC inhibit bacterial growth.MethodsMSC were isolated from the peripheral blood of healthy horses. Gram-negative E. coli and gram-positive S. aureus were cultured in the presence of MSC and MSC conditioned medium (CM), containing all factors secreted by MSC. Bacterial growth was measured by plating bacteria and counting viable colonies or by reading the absorbance of bacterial cultures. Bacterial membrane damage was detected by incorporation of N-phenyl-1-naphthylamine (NPN). Antimicrobial peptide (AMP) gene and protein expression by equine MSC were determined by RT-PCR and Western blot analysis, respectively. Blocking of AMP activity of MSC CM was achieved using AMP-specific antibodies.ResultsWe found that equine MSC and MSC CM inhibit the growth of E. coli and S. aureus, and that MSC CM depolarizes the cell membranes of these bacteria. In addition, we found that equine MSC CM contains AMPs, and blocking these AMPs with antibodies reduces the effects of MSC CM on bacteria.ConclusionsOur results demonstrate that equine MSC inhibit bacterial growth and secrete factors that compromise the membrane integrity of bacteria commonly found in skin wounds. We also identified four specific AMPs produced by equine MSC. The secretion of AMPs may contribute to the value of MSC as a therapy for cutaneous wounds in both horses and humans.

Critical points and the presence of pathogenic bacteria in iced beverage processing lines

Ice can be contaminated by pathogenic bacteria. This study aimed to identify critical points in iced beverage production and distribution lines to examine the presence of pathogenic bacteria in a beverage and its processing environment, as well as when water and ice used as main ingredients. The critical points were determined using the principles of Hazard Analytical Critical Control Point (HACCP) to analyze each processing and distribution step from the survey. Samples collected from the points of concern based on the critical points that were found were tested for pathogens by conventional method and molecular method using primers and polymerase chain reaction (PCR). Escherichia coli was found in 6.34% of samples, and 0.7% of them were confirmed as enterotoxigenic Escherichia coli (ETEC) by PCR. Vibrio cholerae was found in in 0.7% of water samples used to make iced beverages and in ice production, as well as in 2.12% of distribution and production tools. Salmonella Typhimurium was found in 1.4% of water samples used to make ice and ice products. Staphylococcus aureus was found in 2.02% of the surfaces of ice distribution and production tools and in 5.05% of production and distribution workers' hands. S. aureus counts ranged from 2.4×102 - 3.5×102 colonies/100 cm² surface area and 1.9×10¹ - 3.7×102 colonies/workers' hands. Control on many critical points in iced beverage processing and distribution is required so that the beverages are safe for consumption.

Importance of feed structure (particle size) and feed form (mash vs. pellets) in pig nutrition – A review

Pigs are monogastric animals with simple, single-chambered stomach and require easily digestible, high quality feed. One of the most important factors that determine feed utilization by pigs is the particle size distribution. The reduction of particle size of feed improves pigs’ performance due to increased specific surface of feed particles allowing better contact with digestive enzymes. In this respect, fine grinding could be recommended in production of pig feed. Additionally, in modern pig production dry feed is predominantly used in pelleted form, which is mainly due to improved (i.e. decreased) feed conversion ratio (FCR) of pigs fed pelleted feed, but also due to other advantages of pelleted over mash feed.Size of feed particles is strongly reduced during pelleting process. Consequently, digestibility of nutrients in pig feed could be improved. On the other hand, presence of high quantities of fine particles in pig feed (both mash and pelleted) negatively affects the health of gastro-intestinal tract (GIT) leading to higher incidence of stomach ulceration and other negative alterations of gastric mucosa (keratization, erosions). Gastric ulcers are one of the most important causes of sudden death on farm that can result in large economic losses in pig production. Concerning that the animal therapy is expensive, labor-intensive, and mostly non-effective due to late recognition of ulceration, prophylactic recommendations are required. Thus, according to literature data, decreasing the quantity of fine particles in pig feed is strongly recommended.Particle size distribution of the pigs’ feed has a strong influence on presence of pathogen bacteria in GIT of pigs. Feeding pigs with coarse mash feed decreases pH value of stomach content compared to pigs fed finely ground mash feed and compared to pigs fed pelleted feed. This can be explained by slower passage rate, increased dry matter, and more dense consistency of stomach content in pigs fed coarse mash diets. Consequently, feed acidification in stomach is better, number of lactic acid bacteria and concentration of organic acids is higher, and pH of stomach content is lower. These conditions create additional “barrier” against pathogen bacteria.According to available data, optimal particle size of diets for pigs is in the range between 500 and 1600μm, while particles smaller than 400μm are considered as undesirable with high ulcerogenic capacity. Optimal particle size could be designed in the grinding process, and it was shown that the most convenient grinding method is to combine roller mill and hammer mill. Concerning that nowadays pigs are mainly fed pelleted feed, and that pelleting causes strong additional grinding of feed particles, particle size distribution (PSD) obtained within the grinding process would be dramatically changed during pelleting. The possibilities to decrease the intensity of grinding of particles during pelleting, by variation of parameters of pelleting process, are very limited. Modified extrusion process (i.e. processing using expander) followed by shaping element, is suggested in the literature as an alternative for pelleting in order to obtain agglomerated pig feed with preserved PSD, but this process is not extensively studied so far.

Evidence for altered levels of IgD in the nasal airway mucosa of patients with chronic rhinosinusitis

IgD is an enigmatic antibody isotype best known when coexpressed with IgM on naive B cells. However, increased soluble IgD (sIgD) levels and increased IgD+IgM- B-cell populations have been described in the human upper respiratory mucosa. We assessed whether levels of sIgD and IgD+ B cell counts are altered in nasal tissue from patients with chronic rhinosinusitis (CRS). We further characterized IgD+ B-cell populations and explored clinical and local inflammatory factors associated with tissue sIgD levels. sIgD levels were measured by means of ELISA in nasal tissues, nasal lavage fluid, sera, and supernatants of dissociated nasal tissues. IgD+ cells were identified by using immunofluorescence and flow cytometry. Inflammatory mediator levels in tissues were assessed by using real-time PCR and multiplex immunoassays. Bacterial cultures from the middle meatus were performed. Underlying medical history and medicine use were obtained from medical records. sIgD levels and numbers of IgD+ cells were significantly increased in uncinate tissue (UT) of patients with chronic rhinosinusitis without nasal polyps (CRSsNP) compared with that of control subjects (4-fold, P<.05). IgD+ cells were densely scattered in the periglandular regions of UT from patients with CRSsNP. We also found that IgD+CD19+CD38bright plasmablast numbers were significantly increased in tissues from patients with CRSsNP compared with control tissues (P<.05). Among numerous factors tested, IL-2 levels were increased in UT from patients with CRSsNP and were positively correlated with tissue IgD levels. Additionally, supernatants of IL-2-stimulated dissociated tissue from patients with CRSsNP had significantly increased sIgD levels compared with those in IL-2-stimulated dissociated control tissue exvivo (P<.05). Tissue from patients with CRS with preoperative antibiotic use or those with pathogenic bacteria showed higher IgD levels compared with tissue from patients without these variables (P<.05). sIgD levels and IgD+CD19+CD38bright plasmablast counts were increased in nasal tissue of patients with CRSsNP. IgD levels were associated with increased IL-2 levels and the presence of pathogenic bacteria. These findings suggest that IgD might contribute to enhancement mucosal immunity or inflammation or respond to bacterial infections in patients with CRS, especially CRSsNP.