Colony-forming Units Per Gram Research Articles

Fatty Acid Profile and Escherichia coli and Salmonella sp. Load of Wild-Caught Seaweed Fly Fucellia maritima (Haliday, 1838) (Diptera: Anthomyiidae).

World aquaculture is expected to continue to grow over the next few decades, which amplifies the need for a higher production of sustainable feed ingredients for aquatic animals. Insects are considered good candidates for aquafeed ingredients because of their ability to convert food waste into highly nutritional biomass. However, commercially available terrestrial insect species lack n-3 long-chain polyunsaturated fatty acids (LC-PUFAs), which are essential biomolecules for marine cultured species. Nevertheless, several coastal insect species feature LC-PUFAs in their natural fatty acid (FA) profile. Here, we analysed the lipidic profile of wild-caught seaweed fly Fucellia maritima, with a focus on their FA profile, to evaluate its potential to be used as an aquafeed ingredient, as well as to screen for the presence of pathogenic bacteria. Results showed that the flies had a total lipid content of 13.2% of their total dry weight. The main classes of phospholipids (PLs) recorded were phosphatidylethanolamines (PEs) (60.8%), followed by phosphatidylcholine (PC) (17.1%). The most abundant FA was palmitoleic acid (C16:0) with 34.9% ± 4.3 of total FAs, followed by oleic acid (C18:1) with 30.4% ± 2.3. The FA composition of the flies included essential fatty acids (EFAs) for both freshwater fish, namely linoleic acid (C18:2 n-6) with 3.4% ± 1.3 and alpha-linoleic acid (C18:3 n-3) with 3.4% ± 1.9, and marine fish, namely arachidonic acid (C20:4 n-6) with 1.1% ± 0.3 and eicosapentaenoic acid (C20:5 n-3) with 6.1% ± 1.2. The microbiological analysis found 9.1 colony-forming units per gram (CFU/g) of Enterobacteriaceae and no presence of Salmonella sp. was detected in a sample of 25 g of fresh weight. These findings indicate that Fucellia maritima biomass holds the potential to be used as an additional aquafeed ingredient due to its FA profile and the low count of pathogenic bacteria, which can contribute to the optimal growth of fish and shrimp with a low risk of pathogen transfer during the feed production chain.

Effect of Commercial and Autochthonous Bioprotective Cultures for Controlling Listeria monocytogenes Contamination of Pecorino Sardo Dolce PDO Cheese.

The composition and physicochemical characteristics of short-aged Pecorino Sardo PDO (Protected Designation of Origin) cheese makes it permissive to Listeria monocytogenes growth. The PDO product specification stipulates that this cheese is produced with whole sheep's milk inoculated with cultures from the area of origin. Therefore, the use of bioprotective cultures for the inhibition of pathogens in PDO cheeses is allowed only if autochthonous microorganisms are used. Furthermore, bioprotective cultures are generally used on the cheese surface to prevent the outgrowth of L. monocytogenes, the application of which can be time-consuming and require specialist technical knowledge. In this study, we examine the direct addition of bioprotective cultures to the cheese vat and compare the activity of a commercial bioprotective culture (Lactiplantibacillus plantarum) and an autochthonous lactic acid bacterium with bioprotective properties (Lactobacillus delbruekii sups. sunkii), for the inhibition of L. monocytogenes in Pecorino Sardo PDO cheese. Three types of Pecorino Sardo PDO cheese were made with bioprotective cultures added directly to the cheese milk along with the starter inoculum: PSA, with the commercial bioprotective culture; PSB, with the autochthonous bioprotective culture; and a CTRL cheese with no bioprotective culture. A challenge test was performed on each of these cheeses by artificially contaminating the cheese surface with L. monocytogenes (2 Log10 CFU/g). Three batches of each cheese type were analyzed to enumerate mesophilic and thermophilic lactic acid bacteria and to investigate the growth potential of L. monocytogenes during manufacturing, at the end of ripening, at the end of shelf-life, and after 180 days from cheese production. Both bioprotective cultures tested in this study showed inhibitory action against the pathogen with 0.3-1.8 Log10 CFU/g (colony-forming unit per gram) reduction levels. The autochthonous organism, L. sunkii, was as effective as the commercially supplied culture, and the addition of the bioprotective cultures to the cheese-making procedure offered protection against L. monocytogenes. The direct addition of bioprotective cultures to the making procedure of Pecorino Sardo PDO cheese is a potentially innovative strategy to improve the safety of this product.

Fungal spoilage of cayenne pepper (Capsicum annuum) and scotch bonnet pepper (Capsicum chinense)

The fungal spoilage of scotch bonnet pepper and cayenne pepper was investigated. About 20 Samples of cayenne pepper (Capsicum annuum) and scoth bonnet pepper (Capsicum chinense) were obtained from five differerent sellers (four samples each) including two (2) spoiled and two (2) fresh samples in Wadata market in Makurdi metropolis. The samples were serially diluted and dilution 10-2 and 10-4 dilution factors were used, inoculation was done using pour plate method onto SDA medium and the colonies were enumerated in colony forming unit per gram (CFU/g). Identification of the colonies was done from the colony characteristics such as color, shape and type of hyphae under the microscope using the compendium for fungi. Production of afflatoxin by the isolated fungi was also determined. The occurrences of the isolated organisms was recorded based on frequencies and percentage. The isolated fingi were Trychophyton Sp, Geotrichum Sp, Mucor Sp, and Yeast. Preservation and proper handling of different species of pepper should be taken seriously and hygienically to reduce spoilage organisms and hence bring post-harvest losses of pepper to a minimal level.

Regionalized analysis of gut barrier components in the small intestine of BALB/c mice following chronic immobilization stress.

Microbiota and tight junction proteins (TJPs) are components of the gut barrier, and are considered stress targets that have deleterious effects on intestinal homeostasis. This study aimed to evaluate the effects of chronic immobilization stress on selected small intestine homeostasis parameters. Female BALB/c mice were divided into a stress group that underwent short-term immobilization for 2 h per day for 4 consecutive days, and a non-stressed control group (n = 6 per group). Proximal and distal small intestine samples were excised to assess colony-forming units per gram (CFU/g) of total bifidobacteria in selective agar plates, luminal albumin was assessed using immune-enzymatic assay, pro-inflammatory cytokines were evaluated using reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and TJPs (pore-forming, claudin (Cld)-2; pore-sealing, Cld-4; ambiguous, Cld-7, -12 and -15) were assessed with RT-qPCR and western blotting. Compared with the control group, the stress group had lower body weight and energy intake. In the distal region, the stressed mice had lower bifidobacteria count and messenger ribonucleic acid (mRNA) expression of Cld-2, Cld-4 and Cld-12, though they had more albumin and higher interleukin (IL)-6 mRNA expression. Within the proximal region, the stressed mice had higher mRNA expression of tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), IL-6, Cld-7, Cld-12, and Cld-15, along with lower levels of IL-10 and Cld-4. However, mRNA and protein expression of TJPs were discordant. These findings indicate divergent stress-induced outcomes in the small intestine, evidenced by the elicitation of a pro-inflammatory response and decreased anti-inflammatory response in the duodenum, and by increased albumin transudation and decreased bifidobacterial growth in the distal region.

Comparative Evaluation of the Effects of Different Dietary Fibers as Natural Additives on the Shelf Life of Cooked Sausages

Background: The incorporation of dietary fibers in meat and processed meats has been introduced as an approach to amend the nutritional quality and technological properties of the products. Objectives: This study explores the effects of four dietary fibers, including orange fiber (OF), wheat fiber (WF), bamboo fiber (BF), and carrot fiber (CF), on shelf life of emulsion-type cooked sausages. Methods: Microbiological and sensory analyses were performed to evaluate the shelf life of the treated products during 60 days of storage under refrigeration condition (4°C). Results: The results showed that all tested fibers improved the shelf life of the samples. The microbiological examinations revealed that while all the fibers could hinder the growth of spoilage bacteria, OF sample with the mesophilic, psychrotrophic, and lactic acid bacteria counts of 5.95, 4.78, and 5.27 log10 colony-forming unit per gram (CFU/g), respectively showed the highest microbiological quality at the end of the storage. Taste, odor, and overall acceptability of the samples were not significantly affected by the fibers, and texture was the sole sensory attribute that improved in the dietary fiber incorporated products, especially in the OF sample. Conclusions: According to our results, among the various fibers, OF is recommended to the meat industry to extend the shelf life of cooked sausages.

Effects of extrusion specific mechanical energy and dryer conditions on the survival of Bacillus coagulans GBI-30, 6086 for commercial pet food applications

In companion animal nutrition, probiotics (direct-fed microbials) are considered functional ingredients that benefit the gastrointestinal and immune health of the host. Bacillus coagulans GBI-30, 6086 is a spore-forming bacterial strain that has been reported to survive environmental stresses, heat processing, and extreme-pH conditions. Extrusion cooking is the most widely used method to produce commercial dog and cat foods, however the thermal and mechanical forces exerted during extrusion and drying present a challenge for guaranteeing the viability of live microorganisms after processing. Two experiments were conducted to determine the reduction in viability of the microorganism at graded flour inoculation levels (0, 6.2, 6.7, and 7.3 log10 colony forming units per gram (CFU/g)) subjected to extrusion cooking under varied levels of specific mechanical energy that were achieved by adjusting the extruder water input (10, 12, and 20 kg/h), and extruder screw speed (400, 500, and 600 rpm). A second experiment was conducted to determine the survival of the microorganism subjected to three dryer conditions (49 °C for 10 min; 107 °C for 16 min; and 66 °C for 46 min). Enumeration of bacterial colony forming units was performed on pre- and post-processing samples. Extrusion data were analyzed using a general linear model using the GLIMMIX procedure, and dryer data were analyzed as a completely randomized design with one-way analysis of variance (SAS v. 9.4, SAS Institute, Inc., Cary, NC) with significance accepted at a level of 95% confidence (α = 0.05). The results indicate that the low SME extrusion conditions (in-barrel moisture of 35%, extruder screw speed of 400 rpm, and specific mechanical energy of 129 kJ/kg) resulted in the greatest retention (P < 0.05), with a mean log10 reduction of viable spores of 0.44, 2.15, and 2.67 for the low, moderate, and severe extrusion conditions, respectively. Viability of the spores through three dryer conditions were observed to be similar across all treatments. This study also demonstrated that the greatest losses of viability occurred during extrusion rather than drying, and that in-barrel moisture and extruder screw speed are two operational parameters may be modified for the optimization of Bacillus coagulans survival in extruded foods.

Microbial Quality Assurance of Dried Roots of Chlorophytum borivilianum Sant F (Safed Musli)

Aim: To assess total bacterial load and detect E. coli and Salmonella in Chlorophytum borivilianum (Safed Musli), cultivated and processed in the Bundelkhand region.&#x0D; Study Design: Quality assurance of medicinal plant raw material is essential for the preparation of good quality herbal medicines. The dried roots of Safed Musli were collected from different regions of the Bundelkhand and total aerobic microbial count was measured.&#x0D; Methodology: We collected 10 samples of Safed Musli dried roots from different regions of Bundelkhand. A stock suspension was prepared using sterile peptone water and plated on a nutrient agar medium which was used to calculate total aerobic bacterial load as colony-forming units per gram (CFU/g). Further, biochemical tests were performed to confirm the identification of E. coli and Salmonella.&#x0D; Results: The total aerobic bacteria count in dried roots of Safed Musli was within the permissible limit (105-107CFU/g of dried raw material) as per globally recognized pharmacopoeia and other regulatory agencies such as the Central Council of Research in Ayurvedic Sciences (CCRAS), Ministry of AYUSH, Govt. of India, United States Pharmacopoeia (USP), Brazilian Pharmacopoeia (BP), European Pharmacopoeia (EP), World Health Organization (WHO), American Herbal Products Association (AHPA) and National Science Foundation/American National Standards Institute (NSF/ANSI). Although 2 out of 10 samples were found to have contamination of E. coli and Salmonella within the permissible limit of WHO, EP and NSF/ANSI.&#x0D; Conclusion: The results indicated that the Safed Musli cultivated and processed in the Bundelkhand region is suitable for the preparation of herbal medicines and food supplements.

Are Semi-Quantitative Clinical Cultures Inadequate? Comparison to Quantitative Analysis of 1053 Bacterial Isolates from 350 Wounds.

Early awareness and management of bacterial burden and biofilm is essential to wound healing. Semi-quantitative analysis of swab or biopsy samples is a relatively simple method for measuring wound microbial load. The accuracy of semi-quantitative culture analysis was compared to ‘gold standard’ quantitative culture analysis using 428 tissue biopsies from 350 chronic wounds. Semi-quantitative results, obtained by serial dilution of biopsy homogenates streaked onto culture plates divided into 4 quadrants representing occasional, light, moderate, and heavy growth, were compared to total bacterial load quantified as colony-forming units per gram (CFU/g). Light growth, typically considered an insignificant finding, averaged a clinically significant 2.5 × 105 CFU/g (SE = 6.3 × 104 CFU/g). Occasional growth (range: 102–106 CFU/g) and light growth (103–107 CFU/g) corresponded to quantitative values that spanned a 5-log range; moderate and heavy growth corresponded to a range of 4-log and 6-log, respectively, with a high degree of overlap in range of CFU/g per category. Since tissue biopsy and quantitative culture cannot be widely practiced and semi-quantitative analysis is unreliable, other clinically relevant approaches are required to determine wound bioburden and guide best management practices. Fluorescence imaging is a point-of-care technology that offers great potential in this field.

Prevalence of Salmonella spp. and Staphylococcus aureus in Chicken Meat and Pork from Cambodian Markets.

Salmonella spp. and Staphylococcus aureus are two of the most common foodborne bacteria in animal-source foods (ASF) that cause illness worldwide. This study aimed to determine the prevalence of Salmonella spp. and S. aureus in chicken meat and pork in markets in Cambodia. Sampling was done in 52 traditional markets and 6 supermarkets in 25 provinces of Cambodia between October 2018 and August 2019. In total, 532 samples were obtained: chicken meat and pork (n = 408, 204 of each), chicken and pork cutting board swabs (n = 124, 62 of each). All samples were analyzed for the presence of Salmonella spp. and S. aureus; colony-forming units per gram (CFU/g) of coagulase-positive Staphylococci (CPS) were counted, and a subset of samples was also analyzed for the most probable number (MPN, n = 136) of Salmonella. The overall prevalence of Salmonella spp. and S. aureus were 42.1% (224/532) and 29.1% (155/532), respectively, with 14.7% (78/532) of samples containing both bacteria. The prevalence of Salmonella spp. in chicken meat was 42.6%, on chicken cutting board it was 41.9%, on pork it was 45.1%, and the pork cutting board 30.6%. Chicken meat had a significantly (p-value < 0.05) higher prevalence of S. aureus, 38.2%, compared to the chicken cutting board, 17.7%, pork 28.9%, and pork cutting board 11.3%. Mean MPN-Salmonella was 10.6 MPN/g in chicken and 11.1 MPN/g in pork samples. Average Log CFU/g of CPS in chicken and pork samples were 2.6 and 2.5, respectively. The results indicate that chicken meat and pork in Cambodia were highly contaminated with Salmonella spp. and S. aureus, posing risks to consumers’ health. Urgent interventions are necessary to improve hygiene for safer meat in Cambodian markets.

Colloidal Iron Oxide Formulation for Equine Hoof Disinfection.

Simple SummaryHorse hoof possesses a micro-flora that may promote the development of secondary infections following wounds or surgery. For this reason, in vitro and in vivo experiments were used to verify and compare the bactericidal power of two well-known disinfectants, Povidone-iodine (PI) and Chlorhexidine (CHx), and a new stable colloidal suspension of iron oxide, called Iron Animals (IA). In vitro, IA was able to completely suppress the bacterial growth on all of the tested microorganisms. In vivo, PI alone possessed the lowest efficacy for hoof disinfection; CHx resulted as the best treatment in 18% of cases. CHx + IA emerged as the best disinfection protocol for equine hoof achieving the lowest bacterial load in 55% of cases. The addition of IA, after PI or CHx, improved both disinfectants’ effectiveness, leading to the highest bactericidal activity in 82% of cases. IA would deserve a chance as a possible solution in pre-surgical disinfection of the equine hoof, reducing the risk of infection. The presence of bacteria of various origins on horse hoofs enables the onset of infections following trauma or even post-surgical wounds. Thus, the analysis of new antibacterial substances is of fundamental importance. In this study, the antibacterial efficacy of Iron Animals (IA), a stable colloidal suspension of iron oxide, organic acids, and detergents, was tested in vitro and in vivo. In vitro assays were performed to test the unspecific inhibitory effect of IA on both gram-positive and gram-negative bacteria monitoring the microorganism growth by spectrophotometry (optical density OD600) at 37 °C for 24 h. In vivo test consists on the quantification of the bacterial load in colony forming units per gram (CFU/g) of specimens collected from the frog region of the anterior hooves of 11 horses. Sampling followed the application of four disinfectant protocols consisting of two consecutive 3 min scrubs with 50 mL of 10% Povidone-iodine (PI) or 4% Chlorhexidine (CHx), with or without an additional application for 15 min of 10 mL of Iron Animals (PI+IA and CHx+IA). In vitro, IA completely suppressed the bacterial growth of all the tested microorganisms, resulting in effectiveness also against CHx-resistant bacteria, such as Staphylococcus aureus. In vivo, PI emerged as an ineffective protocol; CHx was effective in 18% of cases, but with the addition of IA (CHx + IA) its use emerged as the best disinfectant protocol for horse hoof, achieving the lowest bacterial load in 55% of cases. The addition of IA, after PI or CHx, improves the effectiveness of both disinfectants leading to the highest bactericidal activity in 82% of cases.

Sources of Fungal Contamination of Fresh and Dried Fish in Kisii County, Kenya

Aflatoxins are toxic secondary metabolites produced by Aspergillus flavus and Aspergillus parasiticus that frequently invade foods and feedstuffs. In humid climatic conditions like those found in Kisii County, growth of fungi on dried fish and fish feeds is accelerated due to absorption of moisture from the environment as a result of poor storage and sometimes improper drying. This can lead to creation of favourable conditions for the growth of the toxigenic fungi and consequently production of aflatoxins. This study was conducted to determine the main sources of fungal contamination of selected fresh and dried fish sold in Kisii county, Kenya. Samples of fish were randomly obtained from four main markets in the county. A structured questionnaire was used to determine the sources of fish, mode and hygiene of transportation, handling of fish, duration of storage, hygiene of packaging materials and display environment of fish. Levels of fish contamination were expressed in form of colony forming units per gram (CFU/g) of fish. One-way analysis of variance was used to determine mean differences in parameters analysed. The results revealed that CFU/g of fish from control samples were significantly less than those from fresh and dried fish samples from the various sources (p&lt;0.05). There was generally no significant difference between CFU/g from both fresh and dried fish from the various sources. The study found out that fresh and dried fish sold in Kisii county are contaminated with fungi. There is therefore need for frequent surveillance on the sources of fish contamination and public sensitization of all those involved in fish supply chain. This will help reduce possible aflatoxin contamination thus promoting consumers’ healthy living.

Spray-coating as a novel strategy to supplement broiler feed pellets with probiotic Lactobacillus salivarius NRRL B-30514

Probiotics are potential alternatives to antibiotics in poultry production. In this work, supplementing probiotics on broiler feed pellets (BFPs) was studied for the first time by directly spraying a Lactobacillus salivarius NRRL B-30514 suspension with 10% w/v milk powder and 0, 1, and 5% w/v sucrose, followed by drying at 60 °C for 0–30 min. For suspensions without sucrose, no visual change of BFPs was noticed after spraying the cell suspension for up to 30 s; the coated BFPs dried for a longer time had a lower water activity and lower bacterial viability after preparation; and the coated BFPs dried for 30 min had about 5 log colony-forming unit per gram (CFU/g) viable L. salivarius after 30-day storage at room temperature. Under the optimized conditions (spraying for 30 s and drying at 60 °C for 30 min), supplementing 1% (w/v) sucrose in coating suspension further improved bacterial viability to 6.71 ± 0.14, 5.53 ± 0.24, and 4.28 ± 0.12 log CFU/g after 0, 30, and 730 days, respectively. Coatings slightly increased the diameter but showed no negative influence on the hardness, moisture sorption properties, and surface morphology of BFPs. This approach preventing thermal deactivation of probiotics during pelleting may be significant to supplementing probiotics to animals.

Formulation of a Bio-Packaging Based on Pure Cellulose Coupled with Cellulose Acetate Treated with Active Coating: Evaluation of Shelf Life of Pasta Ready to Eat.

An active packaging based on pure cellulose coupled with cellulose acetate coated with layered double hydroxide (LDH), hosting 4-hydroxybenzoate (listed in EC-Directive 10/2011) as an antimicrobial agent, was formulated and realized. The release of 4-hydroxybenzoate ionically bonded to the LDH layers was much slower than the molecule freely dispersed into the coating. The capability of the active packaging to inhibit Pseudomonas, Escherichia coli, Salmonella and Lactic Bacteria was evaluated, as well as the global migration with three different food simulant (i.e., acetic acid at 3% (v/v), ethanol at 50% (v/v) and vegetable oil) that demonstrated, in compliance with the migration limits of the EU regulation, the suitability of the prepared packaging to be employed as food contact material. Ready to east cooked tomato pasta was packaged into the active trays and in uncoated, as control, up to 30 days at 4 °C. Organoleptic characteristics, mold evolution, total mesophilic aerobic counts (TBC), Enterobacteriaceae, Lactic Bacteria and Pseudomonas, and in colony forming unit per gram (CFU/g), showed a significant activity of 4-hydroxybenzoate in increasing the shelf life of the pasta ready to eat.

Egg Production in Poultry Farming Is Improved by Probiotic Bacteria.

Antimicrobial resistance (AMR) is one of the most serious threats for human health in the near future. Livestock has played an important role in the appearance of antibiotic-resistant bacteria, intestinal dysbiosis in farming animals, or the spread of AMR among pathogenic bacteria of human concern. The development of alternatives like probiotics is focused on maintaining or improving production levels while diminishing these negative effects of antibiotics. To this end, we supplied the potential probiotic Enterococcus faecalis UGRA10 in the diet of laying hens at a final concentration of 108 Colony Forming Units per gram (CFU/g) of fodder. Its effects have been analyzed by: (i) investigating the response of the ileum and caecum microbiome; and (ii) analyzing the outcome on eggs production. During the second half of the experimental period (40 to 76 days), hens fed E. faecalis UGRA10 maintained egg production, while control animals dropped egg production. Supplementation diet with E. faecalis UGRA10 significantly increased ileum and caecum bacterial diversity (higher bacterial operational taxonomic unit richness and Faith’s diversity index) of laying hens, with animals fed the same diet showing a higher similarity in microbial composition. These results point out to the beneficial effects of E. faecalis UGRA10 in egg production. Future experiments are necessary to unveil the underlying mechanisms that mediate the positive response of animals to this treatment.

Red cabbage washing with acidic electrolysed water: effects on microbial quality and physicochemical properties

The effects of acidic electrolysed water (AEW) as ‘green’ technology on the microbiological and physicochemical properties of fresh-cut red cabbages were studied. Fresh-cut red cabbages and artificially inoculated red cabbages with Salmonella typhimurium DT104 were washed with distilled water (DW) and different available chlorine concentrations (ACC) of AEW for different times. AEW treatments significantly reduced the populations of native aerobic bacteria, molds, and yeasts, and artificially inoculated S. typhimurium DT104 compared with the DW-treated and untreated red cabbage samples. The effectiveness of AEW treatments was greatly enhanced with increasing ACC and treatment times. S. typhimurium DT104 were not detected in the washing water that were collected after the red cabbages treated by AEW. The surface colour, pH, and total phenolic contents did not significantly change when the red cabbages were washed with DW and AEW containing 100 mg/L available chlorine for 3 min. The anthocyanin contents and antioxidant activities of red cabbage were significantly reduced by 18.5 per cent for cyanidin, 22.1 per cent for pelargonidin, and 11.2 per cent for 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, however, the impacts on the nutritional benefits of red cabbage were considered as limited and acceptable. The optimal process condition of AEW for washing red cabbage was 100 mg/L ACC for 3 min. In these conditions, most of the native microflora were inactivated, and artificially inoculated S. typhimurium DT104 on the red cabbage were reduced by 40.2 per cent [3.67 log CFU/g (log10 colony-forming units per gram)] and with minimal losses of nutrients and antioxidant activity, as well as no requirement of decontamination treatment on the washing water after AEW treatment.

A TaqMan real-time polymerase chain reaction assay for accurate detection and quantification of Fusarium solani in strawberry plants and soil

Abstract A new TaqMan real-time polymerase chain reaction (qPCR) was developed to accurately detect and quantify the soil-borne fungus Fusarium solani, pathogen to strawberry, in plant and soil samples. The assay was designed using sequences from the translation elongation factor 1 alpha (EF-1α) gene. The assay is highly specific: it specifically detected F. solani isolates when tested against 95 fungal isolates from 27 different species. The detection limit of the assay was 50 fg for F. solani genomic DNA and 102 conidia/g soil. A significant correlation (P = 0.0002) was observed between the amount of genomic DNA of F. solani detected by qPCR and the number of fungal propagules present in artificially inoculated soils. The effectiveness of the assay was validated by comparing it to traditional methods for the detection of F. solani in diseased strawberry plants and pre-planting soils from strawberry fields. A substantial and moderate correlation was found between the qPCR-based and traditional detection methods in diseased plants and soils. The amount of F. solani DNA estimated in roots and crowns of symptomatic strawberry plants ranged from 16 to 190 pg/mg fresh tissue. Inoculum densities in pre-planting soils varied between 3.1 × 102 and 1.3 × 105 colony-forming units per gram (CFU/g) of soil. Effectiveness was also evaluated by assessing the ability of the assay to detect decreasing levels of F. solani populations during biosolarization treatment. Taken together, this novel qPCR assay represents a useful tool for rapid assessment of pre-planting soils and nursery plants to prevent F. solani infection and production losses.

Comparison of extrusion and co-extrusion encapsulation techniques to protect Lactobacillus acidophilus LA3 in simulated gastrointestinal fluids

This study aimed to compare two processes for the encapsulation of Lactobacillus acidophilus LA3 using alginate or a blend of alginate–shellac as the wall material and sunflower oil as the core. The particles were dried and stored for up to 60 days at 25 °C for stability studies, and the survival of the encapsulated probiotics in simulated gastrointestinal fluids was evaluated. The viability of the LA3 encapsulated by co-extrusion using alginate or a blend of alginate–shellac was, respectively, 6.2 and 7.2 log CFU/g (colony-forming units per gram), after 60 days of storage, while the viability of the LA3 encapsulated by extrusion was, respectively, 5.3 and 6.2 log CFU/g. The particles produced with the blend of alginate–shellac presented additional protection to the probiotics, as revealed by the increased survival rates in simulated gastrointestinal fluids. Taken together, these data provide a promising alternative for the incorporation of the probiotic L. acidophilus LA3 into new functional foods.

Efficacy of a Fixed Combination of Tetracycline, Chloramphenicol, and Colistimethate Sodium for Treatment of Candida albicans Keratitis

To evaluate the antifungal activity of a fixed antibiotic combination (AC) containing tetracycline (TET), chloramphenicol (CAF), and colistimethate sodium (CS). In vitro: Candida ATCC and clinical strains were used. The minimum inhibitory concentrations (MICs) of AC and of each antibiotic were determined. Fluconazole (FLC) was tested for comparison. Time-killing curves of selected strains were performed. Ex vivo keratitis: corneas were injected intrastromally with the selected strains. After the injection, corneas were divided into groups of treatments: AC, FLC, or saline. Then, the tissues were analyzed for colony-forming units per gram (CFU/g). Propidium iodide (PI) and MitoTracker (MTR) staining were used to investigate the mode of action. Values of MIC required to inhibit the growth of 90% of organisms for the antibiotics alone were higher than FLC. However, their activity was enhanced when used in combination against Candida yeasts. Time-killing curves showed that at 24 hours, AC reduced the load of both strains of approximately 1 Log10 CFU/g compared with the initial inoculum (P < 0.0001). This effect was also significant versus FLC. In ex vivo, AC was effective in decreasing the loads of both strains by 4 Log10 CFU/g with respect to the control. Moreover, it showed higher activity than FLC against Candida albicans ATCC 10231 (1 Log10 CFU/g, P < 0.01 versus control). PI staining demonstrated that CS changed the membrane's permeability, whereas MTR staining demonstrated that TET or CAF altered mitochondrial function. The cells treated with AC and stained showed both effects. In this study, AC showed antifungal efficacy versus Candida spp.; this activity can be due to the synergistic effects of antibiotics in it.

Microbiological quality of commercially available poultry feeds sold in Bangladesh

The study was conducted aiming at the isolation and identification of pathogens from poultry feed manufactured by four different brands namely A (broiler starter), B (broiler finisher), C (layer starter) and D (layer finisher) sold in retail stores of Rangpur city of Bangladesh. All these samples were collected from four randomly chosen outlets and analyzed by culturing in different culture media such as Nutrient broth (NB), Nutrient agar (NA), Salmonella-Shigella (SS) agar, Eosin methylene blue (EMB) agar, MacConkey agar, Triple sugar iron (TSI) agar slant, Motility, Indole, Urease (MIU) and Saboraud Dextrose agar (SDA) media. The bacterial agents were isolated and examined under light microscope for their gross morphological and conventional biochemical characteristics. The bacteriological analyses were done at the Microbiology Laboratory of Hajee Mohammad Danesh Science and Technology University, Dinajpur during the period of January to June, 2014. Total bacterial colonies of all the samples were counted separately according to the American Public Health Association, using nutrient agar medium for total viable count (TVC), Eosine methylene blue (EMB) agar media for total E. coli count (TEC) and Salmonella-Shigella agar for TSC (total salmonella count). Saboraud Dextrose agar (SDA) media was used for detection of fungus. The virulence effect of the organism present in feed were observed by inoculating the organism in poultry. Recorded result showed that average TVC of feed sample A, B, C and D were 5.45x106, 3.28x105, 5.14x106 and 4.53x105 CFU/gm (colony forming unit per gram) respectively. TEC of feed sample A, B, C and D were recorded 6.25x105, 8.26x103, 5.52x105 and 5.65x104 CFU/gm respectively. TSC of feed sample A, B, C and D were recorded 3.15x104, 2.68x103, 4.46x103 and 1.19x104 CFU/gm respectively. The highest TVC, TEC and TSC were found in broiler starter (feed sample A) and lowest TVC, TEC and TSC were found in broiler finisher (feed sample B). Fungal count was 1.85x105 CFU/ gm in layer finisher (feed sample D) could be as a result of their high pathogenecity as reported by researchers elsewhere. These organisms can cause several poultry and farm animal infections specially mycotoxicosis having public health significance to both human and poultry. The presence of high numbers of E. coli and Salmonella spp. in poultry feed were indicative of poor hygienic practices during manufacture, post process contamination and unsatisfactory transportation and reservation. Therefore reinforce the need for preventive control measures, hygienic handling and processing of feeds to reduce the risk of potential human health hazards.Asian J. Med. Biol. Res. March 2017, 3(1): 52-60

Dietary Effect of Probiotic Bacteria, Bacillus amyloliquefaciens-JFP2 on Growth and Innate Immune Response in Rock Bream Oplegnathus fasciatus, Challenged with Streptococcus iniae

The present study was conducted to evaluate the use of Bacillus amyloliquefaciens as a probiotic source in the diet of rock bream, Oplegnathus fasciatus. A total of 210 fingerlings (25.4 ± 0.13 g) were divided into two groups, three replicates each; control (C) and probiotic (P) group (35×3=105 fish per group). C group was fed a basal diet without probiotic while P group was fed with B. amyloliquefaciens spores at a concentration of 1.4 ×106 colony-forming units per gram (CFU/g) of feed. After 90 days, P group fish showed significant improvements in body weight (BW), weight gain (WG), specific growth rate (SGR) and food conversion ratio (FCR) when compared with C group fish. Also, in bi-weekly assessments, serum protein and glucose level showed a significant increase (p&lt;0.05) in probiotic fed fish compared with the control group. Similar increase in serum antioxidant and lysozyme activity was found in the probiotic fed fish group. However, there was a significant decrease in triglyceride and total cholesterol along with increased alanine aminotransferase (ALT) and aspartate amonotransferase (AST) levels in probiotic diet fed fish compared to the control. After seventy days the fish were challenged with Streptococcus iniae and mortality of the probiotic fed fish was significantly lower (p&lt;0.05) when compared with the control. These results demonstrate the benefit of incorporation of B. amyloliquefaciens as a feed supplement to improve the health status of Oplegnathus fasciatus challenged with Streptococcus iniae.