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Employing affordable and uncomplicated sample preparation techniques to recommend the most efficient antibacterial therapy could help reduce antibiotic-resistant bacteria. This study evaluated the suitability of immunoassays and microbiological assays as alternatives for liquid chromatography/mass spectrometry (LC/MS) in determining plasma tylosin concentrations after intramuscular administration at a dose of 20 mg/kg to both healthy and diseased pigs in clinical veterinary practice. The diseased pigs were confirmed using the target genes Actinobacillus pleuropneumoniae (apxIVA) and Pasteurella multocida (kmt1). The methods showed good linearity, precision, and accuracy. In both healthy and diseased pigs, a significant correlation was observed between LC/MS and the microbiological assay (Pearson correlation coefficient: 0.930, p < 0.001 vs. Pearson correlation coefficient: 0.950, p < 0.001) and between LC/MS and the enzyme-linked immunosorbent assay (ELISA) (Pearson correlation coefficient: 0.933; p < 0.001 vs. Pearson correlation coefficient: 0.976, p < 0.001). A strong correlation was observed between the microbiological assay and the ELISA in both healthy and diseased pigs (Pearson correlation coefficient: 0.911; p < 0.001 vs. Pearson correlation coefficient: 0.908, p < 0.001). A Bland-Altman analysis revealed good agreement between the methods, i.e., 95% of the differences were within the limits of agreement. Therefore, the microbiological assay and the ELISA, which demonstrated sufficient precision and accuracy, can be viable alternatives to LC/MS when it is unavailable.

Durian seeds are waste products from durian fruit that have not been utilized properly. Durian seedflour contain high starch as well as tapioca flour, therefore, durian seed flour can be combined withtapioca flour as a filler for sausage dough, in order to reduce production costs of sausage products. Theaim of this study was to analyze the physicochemical, microbiological and organoleptic properties of beefsausage with the addition of 0%, 25% and 50% durian seed flour from a maximum 30% of the flour usedin making sausages. The results of physical characteristics showed that the addition of durian seed flourhad a significant effect (P 0.05) onother physical characteristics of sausages. The microbiological assay showed that durian seed flour didnot affect the total plate count (TPC) of beef sausages. However, the total plate count of the sausages withthe addition of 25% durian seed flour still met the SNI (Indonesian National Standard) for sausages,instead of sausages with 50% durian seed flour addition. The results of chemical analysis showed that theaddition of durian seed flour had no affect on moisture content, ash content, fat content, protein content,and carbohydrate content of the sausages. The hedonic test values on the parameters of aroma, color,texture and general appearance also showed no significant effect (P> 0.05), except for taste parameters.The hedonic quality test value also showed that the addition of durian seed flour had no different effect(P> 0.05) on the parameters of aroma, color, texture, and general appearance, but had different effect (P

Aminoglycosides (AGs) are chemical substances that exist in the bacteriologic category of traditional antibiotic (AB) therapy. The importance of the determination of AG as has been described in many situations by researchers. Because of the narrow therapeutic ranges of AGs, considerable efforts have been attributed to the analysis of AGs in pharmaceutical preparations, serum, and urine specimens for therapeutic drug monitoring purposes. Residues of ABs in many different cases like environment and human food, causes a major concern, as prolonged exposure to ABs is a serious health hazard, related to both side effects of prolonged use and the risk of developing bacterial resistance to various ABs. The major challenge is finding a sensitive and reliable method to determine AGs in a complex matrix. The microbiological assay was a simple and old method for the determination of AGs. Chromatography and spectroscopy methods are the main instrumental methods for analysis that have been employed for these purposes. Biosensor based instrumental systems have been recently used to determine the AG residues in many cases. Each of these methods has its advantages and disadvantages. This review summarizes different ways (microbiological methods, instrumental methods, and biosensor) for the determination of AGs in all cases. Different databases including PubMed, Scopus, and Web of Science with the words of AGs determination and related words for antimicrobial keywords searched without time limitation.

Ciprofloxacin is a fluoroquinolone antibiotic employed to treat infections. There are over three hundred registered generic brands of ciprofloxacin in Nigeria. There has been an observed marked variation in therapeutic or clinical outcome with change in brands by health professionals and patients. In a previous study of six (6) brands we had reported a high degree of inequivalence. In this study, twenty-nine (29) brands of ciprofloxacin were extensively evaluated against compendia requirements. Microbial sensitivity against two test organisms was conducted. In-vitro drug release and in-vivo simulation were established. Analysis of the data generated indicated that all brands passed qualitative test using TLC and disintegration test, six failed hardness test, one failed friability, nine failed antimicrobial assay and six failed percentage content assay. Conclusions drawn from the study for evidence-based clinical decisionwould include the fact that three brands only were found to be bioequivalent to the innovator brand, Ciproxin; while eight brands were bio-inequivalent using the three models of similarity factor (f2), and difference factor (f1) in two media and dissolution efficiency (DE) in pH 4.5. The three brands (Cipro-All, Cifran And Ciprogem) may confidently be used interchangeably with the innovator brand Keywords: Bioequivalence; Ciprofloxacin Drug quality; Generic substitution; Post-marketing surveillance

Old layers sold for human consumption in six open markets in Abeokuta and Ibadan metropolis of Ogun and Oyo States of Nigeria were analyzed for the presence of residues of streptomycin antibiotic. One hundred and twenty samples comprising of muscle (thigh and breast), liver and kidney tissues were subjected to microbiological assay technique for the detection of residues of the antibiotic. The assay was done on antibiotic medium 5 agar with Bacillus subtilis (ATCC 6633) organism. Results showed that 34.17% of the sampled birds were positive for the presence of streptomycin antibiotic. The concentration of the residues ranged between 0.18μg/g and 0.91μg/g. The mean residue levels recorded among the organs are not significantly different (P>0.05). Some of the muscle samples recorded residue concentration levels higher than the W.H.O recommended maximum residue level for streptomycin antibiotic in muscle tissue of food animals. In view of the importance of antibiotics in the treatment of bacterial diseases in both human and animals, it is advocated that the use of these drugs in food animals be done judiciously. Keywords: Streptomycin, layers

Concerns about the nutritional and sensory qualities of gluten-free (GF) products has generated interest in the evaluation of novel gluten-free ingredients. Folate content is of particular interest due to limited sources of enriched folic acid in a GF diet as well as lack of nutrient composition data in novel flours. The aim of this study was to determine the total folate content and chemical composition of GF flours commercially available in Canada. A tri-enzyme method was used to extract folate from the flour samples, and a microbiological assay was used to measure the total folate contents. The chemical compositions of the GF flours were determined according to standard Association of Official Agricultural Chemists (AOAC) International methods. Compared to all-purpose flour, 265 ± 6.9 μg/100 (dry-weight basis), chickpea flour registered the highest folate content 451 ± 10.8 μg/100 (dry-weight basis) followed by quinoa flour, 174 ± 12.4 μg/ 100 g folate (dry-weight basis). Fonio, had a total starch content of ~77% but was not a source of folate. Flaxseed, chickpea, chia and coconut flours had the highest reported protein contents (mean value: 21.3 ± 1.3%) whereas flaxseed (~42%), and chia (~35%), had the highest lipid content. These findings may inform the selection of gluten-replacement flours with acceptable nutritional properties.

Background Ceftriaxone sodium, an antimicrobial used in parenteral form, does not have a microbiological method by turbidimetry described in the literature. For drugs from antimicrobial class, the existence of a microbiological method for assessing their potency is essential. Not only are the results from the physical-chemical methods enough, but microbiological analyzes are also necessary. Objective and Methods Thus, this paper reports the development and validation of an efficient, accurate, reproducible, fast, and low-cost microbiological assay by turbidimetry to quantify ceftriaxone sodium in powder for injection. Water was used as the diluent to prepare the ceftriaxone solutions. BHI broth as used as culture media for the growth of the S. aureus ATCC 6538 at 9%. Results The method was linear in the range of 100–196 µg/mL, selective against the sample adjuvants and the forced degradation test, precise (intraday RSD 4.53%, interday RSD 3.85% and between analysts tcalculated 0.14 Conclusions and Highlights The turbidimetric method developed in this paper is a convenient and valuable alternative to the routine quality control of ceftriaxone sodium in powder for injection, since it allows a reliable quantification and can be used to complement the physical-chemical analysis.

Microbiologists dread investigating results that are outside of the specifications. However, the investigation must be performed. A decision to reject the batch does not remove the requirement to investigate the failure. Unfortunately, microbiological assay samples are often consumed during the test or the results of the data are obtained several days after the sample is tested. This delay from testing to results often renders the original sample dilution not valid for further testing. Thus, this delay can hinder finding a root cause. Knowing the root cause can aid with microbial control, defining corrective actions, and defining preventative actions that are required to minimize or eliminate the potential for reoccurrence. The root cause is an essential component in understanding the patient impact, status of the product, and regulatory reporting requirements. This article discusses out of specification results obtained from the microbial examination of nonsterile product assays and the subsequent root cause investigation. The focus of this article is on aberrant results obtained from a validated assay with established acceptance criteria.

The detection and assessment of potency of antibiotics are crucial for the pharmaceutics. The valid methods for microbiological assays in pharmacopoeias are mainly based on statistical comparison of the data obtained by measuring the cidal activity resulting from the treatment of the antibiotic active ingredient in the composition of the pharmaceutics with the target microorganism. However, it was seen that there is no validated microbiological method for some active ingredients. Due to microbiological assays are indispensable methods for determining the potency of some active ingredient groups, the calculation of the potency is performed logarithmically. In either turbidimetric or chromatographic methods, the statistical evaluation of the sample is compared with the standard reference material. Analysis data obtained by chromatographic and chemical methods are linear peak areas and spectrophotometer readings. In microbiological methods, the data obtained from the analyzes performed to determine the potency of antibiotics are the inhibition zone diameters or turbidimetric turbidity data. In this study, above-mentioned microbiological assays are compared in the context of the main pharmacopoeias EP, USP, CP, IP and BP, and evaluated in terms of the chromatographic method and classical microbiological method. It has been observed that chromatographic and chemical methods are not available to determine the potency of some pharmaceutical products containing antibiotics. The examinations made reveal the difficulty of analyzing some active ingredient groups according to chemical and chromatographic methods. For this reason, the importance of method validation studies is increasing in order to analyze active substances that do not have alternative analysis methods with microbiological and chemical methods. In this study, all validated microbiological methods were investigated, and it was aimed to determine alternative methods to chromatographic and chemical methods. It was concluded that the realization of new microbiological methods to be validated by evaluating the methods in all differences would facilitate the study. Peer Review History: Received: 18 May 2021; Revised: 15 June; Accepted: 28 June, Available online: 15 July 2021 Academic Editor: Dr. Asia Selman Abdullah, Al-Razi university, Department of Pharmacy, Yemen, asia_abdullah65@yahoo.com UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency. Received file: Reviewer's Comments: Average Peer review marks at initial stage: 6.5/10 Average Peer review marks at publication stage: 7.5/10 Reviewer(s) detail: Dr. Iman Muhammad Higazy, National Research Center, Egypt, imane.higazy@hotmail.com Dr. George Zhu, Tehran University of Medical Sciences, Tehran, Iran, sansan4240732@163.com Similar Articles: AN OVERVIEW ON PHARMACOPOEIAS IN THE WORLD AND MONOGRAPH ELABORATION TECHNIQUES

The purposes of this study were to compare the pharmacokinetic profile of marbofloxacin and antibiotic penetration from blood to milk after intramuscular administration in goats at the beginning, at the peak milk yield and at the end of lactation and to apply the PK/PD modelling approach, so that effective dosage recommendations for this specie could be made. Five healthy lactating goats at 4–9; 25–30 and 150–155 days postpartum were used. At each experience 2 mg/kg intramuscular marbofloxacin was administered. Plasma and milk samples were obtained. Marbofloxacin concentrations were determined by microbiological assay using Klebsiella pneumoniae ATCC 10031 as test microorganism. Plasma pharmacokinetic parameters were calculated from the individual disposition curves, and significant differences between the different moments of lactation were found for AUC (significantly larger for the end of lactation vs the others moments). Milk concentrations were detected for 12 hs postadministration. Our results show that the lactation stage modified the pharmacokinetic profile of marbofloxacin, however these differences are of little consequence from a practical point of view. Volume of milk produced modified milk concentrations of marbofloxacin. A 2 mg/Kg dose level of marbofloxacin would be effective for treating microorganisms with MIC lower than 0.04 μg/mL as E. coli or P. multocida.

Antimicrobial activity of drug products containing antibiotics is often measured using microbiological assays. However, the high values of measurement uncertainty associated with the analytical results obtained from microbiological assays may be an issue to conformity decisions. The aim of this work was to estimate the risk of false decisions in conformity assessment due to measurement uncertainty for the potency of apramycin in pharmaceutical drug products. Monte Carlo method (MCM) simulations were performed in order to estimate global consumers’ (Rc) and producers’ (Rp) risks using a Bayesian approach and specific consumers’ (R′c) and producers’ (R′p) risks using a frequentist approach. Despite of the high value of measurement uncertainty, Rc and Rp were found to be 0.0% and 0.3%, respectively. However, R′c and R′p were found to be high when the analytical result is close to the specification limits. Risk estimation using Bayesian approach is recommended to be applied by manufacturers, while frequentist approach may be an alternative to regulatory and third-party laboratories.

Purpose To investigate the long-term chemical and physical stability of 5-mg/mL acyclovir solution in polypropylene bags stored at 5°C ± 3°C for 2 months in order to determine the feasibility of batch production by a centralized intravenous additive service. Methods Eight empty 100-mL polypropylene bags (bags A) and 8 empty 250-mL bags (bags B) were respectively filled with 60 mL and 200 mL of 5-mg/mL acyclovir and 0.9% sodium chloride injection (NaCl) under aseptic conditions through a semiautomated manufacturing process and vacuum packed before storage at 5°C ± 3°C. Four bags A and 4 bags B were tested for chemical stability via a stability-indicating high-performance liquid chromatography (HPLC) method immediately after preparation (time 0) and after 7, 14, 21, 28, 35, 42, and 63 days. Samples for microbiological assay were collected on days 0 and 63 from 4 bags A and 4 bags B immediately after breaking the vacuum. Osmolality, pH, and physical stability were assessed by visual examination, Subvisible particle counting was performed on 6 additional bags (3 each of bags A and B). Results Mean percentage loss of acyclovir relative to the mean experimental concentration at time 0 was below 5% over the 63-day study period.. No significant differences of pH, no change in color and no precipitate were observed during the study. Subvisible particle counts were compliant with European Pharmacopoeia requirements. Acyclovir solutions remained sterile over the 63 days of the study. Conclusion Extemporaneously prepared acyclovir 5 mg/mL solutions in 0.9% NaCl stored in polypropylene bags were chemically and physically stable over 63 days when stored at 5°C ± 3°C.

Vancomycin is the drug of choice for methicillin-resistant Staphylococcus aureus keratitis and other ocular infections. Vancomycin ophthalmic drops are not commercially available and require compounding. The present study was designed to investigate the stability of vancomycin ophthalmic drops in normal saline, phosphate-buffered saline (PBS), and balanced salt solution (BSS) while stored at room temperature or under refrigeration. Vancomycin ophthalmic drops (50 mg/mL) were aseptically prepared from commercially available intravenous powder using PBS, BSS, and saline. Solutions were stored at room temperature and in a refrigerator for 28 days. The vancomycin stability was tested by a microbiology assay and high-performance liquid chromatography HPLC analysis immediately after formulation and at days 7, 14, and 28 after storage at room temperature or under refrigeration. The pH, turbidity was also tested. Vancomycin formulations in PBS, BSS and normal saline had initial pH of 5, 5.5, 3 respectively. The formulation in PBS developed turbidity and a slight decrease in pH upon storage. Microbiological assay did not show any change in zone of inhibition with any of the formulation upon storage either at room temperature or under refrigeration. HPLC analysis did not detect any decrease in vancomycin concentration or the accumulation of degraded products in any of the formulations upon storage either at room temperature or under refrigeration. Vancomycin ophthalmic drops prepared using PBS, BSS, and normal saline were stable up to the tested time point of 28 days, irrespective of their storage temperature.

Associated with the use of removable prostheses, the development of candidiasis, called prosthetic stomatitis, is frequently observed. In view of the application of silver nanoparticles (AgNP) in dentistry that may offer antimicrobial effect, the aim of this study was to evaluate the effect of adding AgNP with different concentrations during thermopolymerization or immersion of acrylic in this substance in the properties antifungal, mechanical and cytotoxic. The groups were divided: addition of 1% silver nanoparticle solution (G1), addition of 2.5% silver nanoparticle solution (G2), addition of 5% silver nanoparticle solution (G3), immersed for 10 min in aqueous silver nanoparticle solution (G4), immersed for 24 hours in aqueous silver nanoparticle solution (G5). In the cytotoxicity assay, at all evaluation times, all groups showed cytotoxic effect (p

BACKGROUND: Vitamin B6 insufficiency has been linked to increased risk of cancer and other chronic diseases. The circulating concentration of pyridoxal 5'-phosphate (PLP) is a commonly used measure of vitamin B6 status. Ratios of substrates indicating PLP coenzymatic function and metabolism may be useful complementary measures to further explore the role of vitamin B6 in health. OBJECTIVES: We explored the sensitivity of 5 outcomes, namely PLP concentration, homocysteine:cysteine (Hcy:Cys), cystathionine:cysteine (Cysta:Cys), the 3´-hydroxykynurenine ratio (HKr), and the 4-pyridoxic acid ratio (PAr) to vitamin B6 intake as well as personal and lifestyle characteristics. MEDTHODS: Dietary intake and biomarker data were collected from participants from 3 nested case-control studies within the European Prospective Investigation into Cancer and Nutrition (EPIC). Bayesian regression models assessed the associations of the 5 biomarker outcomes with vitamin B6 intake and personal and lifestyle covariates. Analogous models examined the relations of Hcy:Cys, Cysta:Cys, and HKr with PLP. RESULTS: In total, 4608 participants were included in the analyses. Vitamin B6 intake was most strongly associated with PLP, moderately associated with Hcy:Cys, Cysta:Cys, and HKr, and not associated with PAr (fold change in marker given a doubling of vitamin B6 intake: PLP 1.60 [95% credible interval (CrI): 1.50, 1.71]; Hcy:Cys 0.87 [95% CrI: 0.84, 0.90]; Cysta:Cys 0.89 [95% CrI: 0.84, 0.94]; HKr 0.88 [95% CrI: 0.85, 0.91]; PAr 1.00 [95% CrI: 0.95, 1.05]). PAr was most sensitive to age, and HKr was least sensitive to BMI and alcohol intake. Sex and menopause status were strongly associated with all 5 markers. CONCLUSIONS: We found that 5 different markers, capturing different aspects of vitamin B6-related biological processes, varied in their associations with vitamin B6 intake and personal and lifestyle predictors. ispartof: AMERICAN JOURNAL OF CLINICAL NUTRITION vol:114 issue:1 pages:338-347 ispartof: location:United States status: published

Danofloxacin is a veterinary fluoroquinolone used to treat respiratory and gastrointestinal diseases of birds, pigs and cattle. The literature reviewed shows some analytical methods to quantify this fluoroquinolone, but microbiological and biological safety studies are limited. The analytical methods were validated by the Official Codes. The LC-DAD method was developed and validated using an RP-18 column, mobile phase containing a mixture of 0.3% triethylamine (pH 3.0) and acetonitrile (85:15, v/v). The microbiological assay was performed by agar diffusion method (3 x 3) and Staphylococcus epidermidis as a microorganism test. Forced degradation studies were performed in both methods. The minimum inhibitory concentration (MIC) was performed by test microdilution and toxicity studies were evaluated using in silico study, cell proliferation, cell viability test, micronuclei and comet assay. LC and a microbiological assay proved linear, accurate, precise, and robust to quantify danofloxacin, but only the LC method showed selectivity to quantify the drug in the presence of its degradation products. These results demonstrate that the LC method is suitable for stability studies of danofloxacin, but a microbiological assay cannot be used to quantify the drug due to the biological activity of the photoproducts. Ex-vivo cytotoxicity and theoretical and experimental genotoxicity were also observed.

Foodborne diseases caused by pathogens and toxins are a serious threat to food safety and human health; thus, they are major concern to society. Existing conventional foodborne pathogen or toxin detection methods, including microbiological assay, nucleic acid-based assays, immunological assays, and instrumental analytical method, are time-consuming, labor-intensive and expensive. Because of the fast response and high sensitivity, cell-based biosensors are promising novel tools for food safety risk assessment and monitoring. This review focuses on the properties of mammalian cell-based biosensors and applications in the detection of foodborne pathogens (bacteria and viruses) and toxins (bacterial toxins, mycotoxins and marine toxins). We discuss mammalian cell adhesion and how it is involved in the establishment of 3D cell culture models for mammalian cell-based biosensors, as well as evaluate their limitations for commercialization and further development prospects.

Experiment was performed on six healthy Marathwadi buffalo calves of either sex of age above 6 months and weighing between 80 to 120 kg to study the different pharmacokinetic parameters after single intravenous administration @ 2 mg/kg body weight by microbiological assay technique. After intravenous administration of the drug, blood samples (4 ml each) of buffalo calves were collected from external jugular vein using disposable needles in clot activator tubes at different time intervals. The schedule of blood collection for pharmacokinetic studies after intravenous administration was at 0, 5, 10, 15, 30 min and 1, 2, 4, 6, 8, 10, 12 and 24 hrs. The serum levels of cefquinome were estimated by microbiological assay technique using large glass plate. Different Pharmacokinetic parameters were calculated as described by different scientists. The peak serum concentration, elimination half life, volume of distribution, total body clearance, absorption half life and area under curve values found were 1.74 ± 0.151 mcg/ml at 2.5 min of sampling time, 1.97±0.14 h, 3.97 ± 0.83 L/kg (Vd(B)) and 2.86 ± 0.34L/kg (Vdss), 1.11 ± 0.13 L/kg.h−1, 0.10 ± 0.02 h, and 1.93 ± 0.23 μg/ml/hr respectively. The bioavailability of cefquinome in buffalo calves was found to be 100 %. It may be concluded that the elimination half life of cefquinome was 2.54 h in Marathwadi buffalo calves indicating the repeating of doses at 12 to 15 h intervals in Marathwadi buffalo calves and the loading dose would be double than the maintenance dose of cefquinome after intravenous administration.

A simple, inexpensive and efficient agar diffusion bioassay, applying cylinder plate method, for determination of josamycin (JOS) in pure and pharmaceutical dosage forms was studied. Using a strain of Micrococcus luteus ATCC 9341 as the test organism, JOS at concentrations ranging from 0.1 to 0.5 µg/mL could be measured. The calibration graph was linear with good correlation coefficient (r = 0.999), and the method validation showed that it was specific and robust. The interference effects of degradation products and excipients were investigated in order to assess the method selectivity. The proposed method was successfully applied to estimate JOS content in tablets and suspension with good precision (intra-day RSD=0.87%; inter-day RSD=1.25%) and accuracy; the recovery values were between 97.74 and 101.13%. Bioassay was correlated with UV spectrophotometric and high performance liquid chromatographic techniques. The results obtained by these three methods were statistically analyzed by analysis of variance (ANOVA), which indicated that there was no significant difference among these methods. Therefore, we recommend this microbiological assay for drug analysis in routine quality control of josamycin in pharmaceuticals as an alternative to the physicochemical methods reported in the literature.

O Brasil é um dos mais importantes produtores de carne bovina do mundo, a produção em 2019 foi de 1,84 milhões de toneladas, o que gerou em receita o valor de US$ 7,59 bilhões, isso representa cerca de 3% das exportações do país. Há uma demanda crescente para o comércio externo de carnes resfriadas, devido a manutenção das características principalmente de maciez e suculência, porém, hoje, o prazo usual da vida de prateleira desses cortes é em torno de 90 dias. Dessa forma conhecer a dinâmica microbiana e físico química em cortes cárneos embalados a vácuo visando o aumento da vida de prateleira, viabiliza a redução de perdas comerciais por deterioração e pode ser uma ferramenta para embasar tecnicamente empresas interessadas ao comércio de carne bovina resfriada para exportação. Assim, o objetivo desse trabalho foi realizar o estudo da vida de prateleira de cortes resfriados bovinos durante 120 dias, através da avaliação de análises físico químicas e microbiológicas. Os cortes avaliados foram o coxão mole e o acém. Foram encaminhados a laboratório externo (credenciado pelo MAPA) 6 peças com peso médio de 1kg, de cada um dos cortes (12 amostras no total) nos períodos: 3, 45, 70, 90, 110 e 120 dias. As características físico-químicas avaliadas foram acidez titulável, pH e índice de peróxidos; e as microbiológicas: coliformes termotolerantes, Clostridium Sulfito-redutor; pesquisa de Salmonella spp; Contagem Padrão de Microorganismos Aeróbios Mesófilos e contagem de Staphylococcus. As análises físicoquímicas realizadas indicaram que todas as amostras apresentaram resultados dentro dos padrões de acordo com a literatura disponível e dentro dos padrões para pH conforme legislação nacional. Não há especificação descrita em normativas oficiais para os demais parâmetros físico-químicos analisados. As análises microbiológicas apresentaram resultados dentro dos padrões estabelecidos pela legislação quando aplicável e os micro-organismos termotolerantes apresentaram resultados dentro dos padrões pela legislação anterior vigente, pois a atual RDC nº 331 de 23 de dezembro de 2019 não determina valores específicos para este grupo de micro-organismos, bem como para a contagem de Staphylococcus coagulase positiva que também apresentou resultados dentro dos parâmetros da legislação anteriormente vigente. A legislação nacional é deficiente em relação às especificações de tolerâncias aceitáveis ou não aceitáveis para muitos micro-organismos presentes em alimentos. Brazil is one of the most important beef producers in the world, production in 2019 was 1.84 million tons, which generated revenue of US $ 7.59 billion, this represents about 3% of exports from the country. There is a growing demand for the external trade of chilled meat, due to its freshness and maintenance of the characteristics mainly of tenderness and juiciness, however, today, the usual shelf life for these cuts is around 90 days. For beef chilled vacuum-packed this period is considered short, considering slaughter, processing, transport to the buyer country until exposure in the consumer market. In this way, knowing the microbial dynamics and chemical physics in vacuumpacked meat cuts in order to increasing shelf life, enables the reduction of commercial losses due to deterioration and can be a way to technically support companies willing to trade chilled beef for export. Thus, the aim of this work was to carry out the study of the shelf life of chilled cuts of beef in the period of 120 days, through the evaluation of physical chemical and microbiological analyzes. The evaluated cuts were chuck and topside. Six pieces with an average weight of 1 kg were sent to the external laboratory, these were sent in pairs to the accredited lab by MAPA in the period’s: 3, 45, 70, 90, 110 and 120 days. The physicochemical characteristics evaluated were titratable acidity, pH and peroxide index; and microbiological ones: count of thermotolerant coliforms, Clostridium Sulphite reducer; Salmonella spp. research, Standard Count of Aerobic mesophiles Microorganisms and count of coagulase-positive Staphylococcus. The physical-chemical analyzes performed indicated that all samples presented results within the standards according to the available literature and within the pH standards according to national legislation. There is no specification described in official regulations for the others analyzed physical-chemical parameters analyzed. The microbiological analyzes showed results within the standards established by the legislation when applicable and the thermotolerant microorganisms showed results within the standards by the previous legislation in force, since the current RDC No. 331 of December 23, 2019 does not determine specific values for this group of micro - organisms, as well as for the counting of positive coagulase Staphylococcus, which also showed results within the parameters of the legislation previously in force. National legislation is deficient in terms of acceptable or unacceptable tolerance specifications for many microorganisms in food.

Background: Hospital-acquired infections (HAIs) remain a common problem, which suggests that standard decontamination procedures are insufficient. Thus, new methods of decontamination are needed in hospitals. Methods: We assessed the effectiveness of a no-touch automated disinfection (NTD) system in the decontamination of 50 surfaces in 10 hospital rooms. Contamination of surfaces was assessed with a microbiological assay and an ATP bioluminescence assay. Unacceptable contamination was defined as &gt, 100 colony forming units/100 cm2 in the microbiological assay, and as &ge, 250 relative light units in the ATP assay. Results: When measured with the microbiological assay, 11 of 50 surfaces had unacceptable contamination before NTD, and none of the surfaces had unacceptable contamination after NTD (p &lt, 0.001). On the ATP bioluminescence assay, NTD decreased the number of surfaces with unacceptable contamination from 28 to 13, but this effect was non-significant (p = 0.176). On the microbiological assay taken before NTD, the greatest contamination exceeded the acceptable level by more than 11-fold (lamp holder, 1150 CFU/100 cm2). On the ATP bioluminescence assay taken before NTD, the greatest contamination exceeded the acceptable level by more than 43-fold (Ambu bag, 10,874 RLU). Conclusion: NTD effectively reduced microbiological contamination in all hospital rooms. However, when measured with the ATP bioluminescence assay, the reduction of contamination was not significant.

The information required for an adequate analytical method validation for either in vitro or in vivo biopharmaceutical studies is discussed in this article. The most commonly applied assay parameters accuracy, precision, detection limit, specificity, selectivity, calibration linearity or nonlinearity, stability, recovery, reproducibility and repeatability are carefully defined along with some recommendations required for assessing data acceptability arrived at the conference on Analytical Methods Validation. Though more specific analytical issues concerning microbiological assay radioimmunoassay and stereoisomer assay were not examined, this article should provide good guidance to pharmaceutical chemists to properly perform analytical method validation and carry out routine analysis.

In 2009, the Australian government mandated the addition of folic acid to bread flour to reduce the incidence of neural tube defects (NTD)-affected pregnancies. In 2011–2012, the Australian Health Measures Survey (AHMS) reported a mean red blood cell (RBC) folate in women of reproductive age (16–44 y) of 1647 nmol/L. Over 99% of women had an RBC folate ≥ 906 nmol/L, a concentration consistent with a very low risk of NTDs if a woman became pregnant. However, RBC folate was measured using an immunoassay, which is not a recommended method due to questionable accuracy. The microbiological assay is the preferred method for RBC folate measurement. To determine whether the immunoassay method may have led to spurious conclusions about the folate status of Australian women, we collected fasting blood samples from 74 healthy non-pregnant, non-lactating women (18–44 y) and measured RBC folate using both the immunoassay and microbiological methods. Mean RBC folate (95% confidence interval) concentration measured with the immunoassay method was 1735 (1666, 1804) nmol/L compared with 942 (887, 1012) nmol/L using the microbiological method. No woman had an RBC folate < 906 nmol/L using the immunoassay method, whereas 46% of women had an RBC folate < 906 nmol/L using the microbiological method. The NTD risk was estimated to be 0.06% using the immunoassay method and 0.14% using the microbiological method. RBC folate using AHMS survey may have underestimated NTD risk in Australian women.

Background Indonesian Ministry of Health advocate doctors, especially in government-owned healthcare facility, to prescribe generic drugs including amoxicillin. Although BPOM (the National Agency of Drug and Food Control) already guarantees that the generic amoxicillin and the branded one were interchangeable, lack of confidence in generic drugs still remains among patients, pharmacists, and doctors. This issue supported by lack of publication confirmed the therapeutic equivalence of branded and generic drugs. This study aims to evaluate and compare the in vitro microbiological assay of different generic and branded amoxicillin that are available in Indonesian market, especially those used in government-owned healthcare facilities. Methods Microbiological assays for five samples of amoxicillin tablet containing 500 mg amoxicillin available in Indonesia were determined using a method from Indonesia Pharmacopeia. Samples were coded as Products A to E. The assay was carried out by measuring the diameter of the inhibition zones in the plate agar incubated with Escherichia coli and Staphylococcus aureus. The obtained data were evaluated to determine the sample potency and compared with the amoxicillin reference standard. Results Minor and insignificant differences (p > 0.05) were found in the diameters of the inhibition zones. Potency ratio measured both in E. coli and S. aureus were all between 95% and 105%. The lowest of the tested samples were from Product C, which resulted to ratio potencies of 96.3% and 95.5% in E. coli and S. aureus, respectively. Conclusions All five samples were in the range of the acceptance criteria. Therefore, from the view of the microbiological assay, these products are in equivalence in quality and are interchangeable.

*Bilican, İsmail ( Aksaray, Yazar ) *Bahadır, Tolga ( Aksaray, Yazar ), Pseudomonas aeruginosa is a pathogenic bacterium in fresh water supplies that creates a risk for public health. Microbiological analysis of drinking water samples is time consuming and requires qualified personnel. Here we offer a screening system for rapid analysis of spring water that has the potential to be turned into a point-of-need system by means of simple mechanism. The test, which takes 1 h to complete, electrically interrogates the particles through a microfluidic chip suspended in the water sample. We tested the platform using water samples with micro beads and water samples spiked with P. aeruginosa at various concentrations. The mono disperse micro beads were used to evaluate the performance of the system. The results were verified by the gold standard membrane filtration method, which yielded a positive test result only for the P. aeruginosa spiked samples. Detection of 0–11 k bacteria in 30 μL samples was successfully completed in 1 h and compared with a conventional microbiological method. The presented method is a good candidate for a rapid, on-site, screening test that can result in a significant reduction in cost and analysis time compared to microbiological analyses routinely used in practice.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) O Brasil é o segundo maior produtor de celulose do mundo, tornando o setor de árvores plantadas de grande importância para a economia nacional. Porém essas indústrias ocupam o sexto lugar em termos de poluição ambiental, sendo responsáveis por descartar efluentes com alto potencial poluidor se não tratados adequadamente. A maioria das indústrias utilizam tratamentos biológicos. O reator biológico de leito móvel (MBBR) vem ganhando popularidade devido sua estabilidade, tamanho compacto, eficiência na remoção de matéria orgânica, além da baixa produção de lodo. Esse sistema permite o crescimento de biomassa aderida (biofilme) em suportes que, com auxílio da aeração, movimentam-se por todo volume do reator. Nesse estudo foi avaliado o desempenho do meio suporte esponjoso Aquaporousgel (APG), desenvolvido pela empresa japonesa Nisshinbo Chemical Inc., adicionado em MBBR empregado no tratamento de efluente de indústria de celulose. O conjunto MBBR-APG foi construído em escala de bancada com volume útil de 1 L e operado por 240 dias com cargas orgânicas volumétricas (COV) de 0,6 e 1,2 kgDQO m-3 d-1, resultando em tempos de detenção hidráulica (TDH) variando de 1,0 a 2,7 dias. A eficiência do tratamento foi determinada pela remoção de demanda química de oxigênio (DQO), demanda bioquímica de oxigênio (DBO5), carbono orgânico total (COT), cor, turbidez, compostos fenólicos totais (CFT), compostos específicos derivados de lignina e também foi avaliada a matriz de excitação e emissão de fluorescência (MEEF). O desenvolvimento do biofilme foi avaliado com a quantificação dos sólidos aderidos, obtenção de imagens em microscopia eletrônica de varredura (MEV) e análises microbiológicas. O MBBR-APG apresentou melhor desempenho na COV 1,2 kgDQO m-3 d-1, com remoções médias de matéria orgânica de 62,7, 48,4 e 55,7% para DBO5, DQO e COT, respectivamente. Mas o tratamento biológico gerou incrementos de outros compostos fluorogênicos. Discute-se a hipótese desse fenômeno ser resultante da ação de microrganismos anaeróbios presentes em microzonas anóxicas formadas no interior do meio suporte esponjoso devido à dificuldade na difusão de oxigênio. As análises em MEV indicaram que o material esponjoso estudado apresenta estrutura funcional superior às tradicionais esponjas de poliuretano, sendo possível observar o desenvolvimento de uma matriz de biofilme composta por resíduos remanescentes do efluente, substâncias poliméricas extracelulares (EPS) e a presença de fungos e bactérias. As análises microbiológicas confirmaram a presença de fungos do gênero Aspergillus e Trichoderma e bactérias Bacillus sp., Serratia sp. E Lysibacillus sp. O trabalho apresentou importantes contribuições sobre o uso de meios suportes esponjosos, biotransformação de compostos em tratamentos biológicos de efluentes de celulose e o desenvolvimento da matriz de biofilme nesses sistemas. Brazil is the second-largest pulp producer in the world and, therefore, the tree sector is of great importance to the national economy. However, these industries are in sixth place in terms of environmental pollution, being responsible for disposing of effluents with high polluting potential if not properly treated. Most pulp mill industries use biological treatments. The moving bed biofilm reactor (MBBR) has been gaining popularity due to its stability, compact size, removal efficiencies, and low sludge production. This system allows the growth of adhered biomass (biofilm) in supports. The supports, also called carriers, move throughout the reactor volume with the aid of aeration. This study evaluated the performance of the Aquaporousgel (APG), a functional sponge developed by the Japanese company Nisshinbo Chemical Inc., used in an MBBR to treat pulp mill effluent. An MBBR system in a laboratory-scale with utile volume of 1 L was filled by this innovating sponge carrier to treat effluent from a pulp industry. The MBBR-APG operated continuously for 240 d with an organic load rate (OLR) of 0.6 and 1.2 kgDQO m-3 d-1, resulting in hydraulic retention times (HRT) ranging from 1.0 to 2.7 days. The treatment efficiency was determined by the removal of chemical oxygen demand (COD), biochemical oxygen demand (BOD5), total organic carbon (TOC), color, turbidity, total phenolic compounds, lignin derivatives, and fluorescence excitation-emission matrix. Biofilm development was evaluated by quantifying adhered solids, scanning electron microscopy (SEM), and microbiological analyses. The MBBR-APG presented a better performance at OLR of 1.2 kgDQO m-3 d1, achieving average organic matter removals of 62.7, 48.4 and 55.7% for BOD5, COD and TOC, respectively. Despite that, the biological treatment generated increments of other fluorogenic compounds. The hypothesis of this phenomenon is the result of the action of anaerobic microorganisms present in anoxic microzones formed in the inner of the sponge carrier due to the difficulty in oxygen diffusion. The SEM analysis indicated that the APG sponge has a functional structure better than the traditional polyurethane sponges. It is possible to observe the development of a biofilm matrix composed of extracellular polymeric substances (EPS) and the presence of fungi and bacteria. Microbiological analyses confirmed the presence of fungi genus Aspergillus and Trichoderma, and bacterial genus Bacillus sp., Serratia sp. And Lysibacillus sp. This study presented important contributions to the use of sponge carrier, compounds biotransformation in biological treatments of pulp effluents, and the development of biofilm matrix in these systems.

BACKGROUND: Harmonizing critical reagents for the folate microbiological assay (MBA) may improve among-laboratory comparability. OBJECTIVE: We assessed the comparability of the MBA for serum folate (S-FOL) and whole-blood folate (WB-FOL) in an international comparison study. METHODS: Eight laboratories obtained a kit containing CDC microorganism inoculum (chloramphenicol-resistant Lactobacillus rhamnosus), CDC calibrator (5-methyltetrahydrofolate), and 23 serum and WB hemolysate samples each. Laboratories analyzed the samples in single measurement over 2 d using 4 conditions: in-house microorganism and in-house calibrator (IH-MO & IH-CAL), in-house microorganism and CDC calibrator (IH-MO & CDC-CAL), CDC microorganism and in-house calibrator (CDC-MO & IH-CAL), and CDC microorganism and CDC calibrator (CDC-MO & CDC-CAL). We calculated geometric mean concentrations for each laboratory and condition and compared data to the CDC MBA (target). RESULTS: The among-laboratory arithmetic mean S-FOL concentrations for the 4 conditions were 30.2, 28.1, 30.0, and 29.9 (group1, 5-methyltetrahydrofolate IH-CAL) compared with 35.3, 33.3, 33.6, and 30.7 nmol/L (group 2, folic acid IH-CAL), respectively; and 428, 405, 398, and 393 (group 1) compared with 469, 423, 477, and 418 nmol/L (group 2), respectively, for WB-FOL. Differences to the CDC MBA target values were smaller for group 1 (range across conditions; S-FOL: 9.9–21%; WB-FOL: 9.0–18%) compared with group 2 laboratories (S-FOL: 13–30%; WB-FOL: 16–32%) and smaller when CDC reagents were used compared with in-house reagents (S-FOL: 12% compared with 22%; WB-FOL: 13% compared with 25%). A linear mixed model estimated a small microorganism effect (S-FOL: 2.3%; WB-FOL: 2.3%) and a larger mean calibrator effect; folic acid compared with 5-methyltetrahydrofolate calibrator produced 12% higher S-FOL and 15% higher WB-FOL results. When laboratories used CDC reagents, the estimated among-laboratory variability was ~10% for S-FOL and WB-FOL. CONCLUSION: Harmonizing the calibrator and microorganism for the folate MBA has the potential to improve the among-laboratory comparability in future surveys.

The ability of folate-producer strains of Streptococcus thermophilus to accumulate folates and the expression of two target genes (folK and folP), involved in the folate biosynthesis, were studied during milk fermentation. An over-expression of folK took place only in the early phase of growth, whereas folP was mainly expressed in the mid log-phase of growth and declined thereafter. The accumulation of total folates, which was quantified by a microbiological assay, was strain-dependent. Two major forms of folates, i.e. tetrahydrofolate (THF) and 5-methyl-tetrahydrofolate (5-Met-THF), were identified and quantified by HPLC. With respect to the level accumulated by a weak folate producer (St 383), used as calibrator in the expression experiments and as control in folate quantification in milk, the strains St 563 and St 399 produced 5-Met-THF in amounts significantly higher than THF. The possibility of using selected folate-producer S. thermophilus strains as functional cultures for a bio-fortification of dairy products is discussed.

Soy protein (tofu) is the most inexpensive source of high nutritional quality protein. The biological value of soy protein is higher than that of casein. The study aimed to prepare croissant with spreadable tofu blends to enhance nutritional value and function. Spreadable tofu blends, with different flavors, were added to the croissant as filling materials. Seven samples were prepared. Samples included; control without addition, and with spreadable tofu only, tofu with chopped green olives, tofu with ground black pepper and chopped green peppers, tofu with sugar; tofu with guava pulp, and tofu with peach pulp. The croissant samples were subjected to chemical, physiological, microbiological and sensory assessment. Results indicated slight differences among treatments within the range of moisture. The protein, fat and ash contents in all the croissant samples stuffed with tofu were higher than the control sample. In contrast, control croissant showed higher carbohydrates content compared to the other samples. Water activity was decreased with extending storage time and addition of tofu mixtures. The sensory test of the croissant samples confirmed that the croissant samples with the tofu blends were more acceptable than the control sample and proved the superiority of the green olives sample and the peach sample among the formulated ones. The microbiological assay assured the safety of all samples. Key words: croissant, spreadable tofu, chemical composition, water activity, sensory evaluation, microbiological examination.

fruit and vegetable wastes and their by-products are formed in great amounts during industrial processing which presents a serious problem to environment. So they need to be managed or prepared to be utilized. The aim of the present investigation is to study the possibility of white watermelon rind powder (WMRP) utilization in manufacturing cake by partial substitution of wheat flour at different levels (10, 20, 30 and 40%).The effect of white watermelon rind powder substitution on the physicochemical, sensory properties and shelf life of the cakes was investigated. Results showed that fiber content of the produced cakes was increased by increasing the substitution levels of WMRP. Minerals content particularly Ca, K and Mg, content was increased in WMRP cake samples. Substitution with WMRP significantly improved the cake volume. Results of textural analysis showed that watermelon rind powder decreased all textural parameters. Regarding to color character, cakes became darker as the level of substitution increased. The sensory evaluation results showed that cakes prepared with WMRP had high mean scores for overall acceptability except in case of 40% substitution level. All WMRP cake samples exhibited a lower microbiological count than control at zero time and during storage periods. So, the substitution with WMRP in cakes processing led to prolong the shelf life of cake compared with control sample with respect to microbiological assay.

Vitamin B12 plays a key role in human biological functions and is vital in the neurological development of infants. The assessment of the vitamin B12 intake in exclusively breastfed babies depends on the reliability of its determination in milk. In this report, we present a new accurate and robust method for quantification of vitamin B12 in human milk. A highly specific sample preparation is applied, associated with chromatographic separation and detection by ICP-MS. Excellent sensitivity and accuracy are reported, with recovery values well within acceptability limits (80-120 %), within- and between-day variability are lower than 10 % and 15 % respectively. Strong correlation with a microbiological assay was observed (r2=0.9) within the validation range (40-1000 pmol/L corresponding to 54 to 1355 ng/L). The method can be used to routinely monitor vitamin B12 in clinical or population observational studies, determine infant’s intake or assess efficacy of mother’s supplementation.

The study objectives were to measure folate content in commonly consumed Korean vegetable dishes prepared outside the home (VDOHs) and to compare the analyzed values with the values calculated using the recipes of a nutrient analysis program. A total of 56 VDOHs were collected from restaurants and catering companies between 2013 and 2016. Folate content was measured by microbiological assay after trienzyme extraction and calculated using the Computer-Aided Nutritional Analysis Program developed by the Korean Nutrition Society. Folate content ranged from 3.1 μg/serving (seasoned bracken, pickled cucumber) to 84.8 μg/serving (curled mallow soybean paste soup) by analysis and from 0.2 μg/serving (pickled cucumber) to 164.3 μg/serving (spinach soybean paste soup) by calculation. The calculated folate values per serving as well as per 100 g were significantly higher than the analyzed values, and the Bland-Altman plots showed poor agreement between the calculated and analyzed values. Our results indicate that folate intake from VDOHs or cooked vegetables could be overestimated by calculation. Therefore, a folate database should be constructed to reflect the weight changes and folate retention of food employing various cooking methods to accurately estimate folate intake in the South Korean population.

For this study, 12 goats of either sex were selected. A group of six goats was administered with cefquinome @ 2 mg/kg by intramuscular route and another group of six goats was administered with cefquinome @ 2 mg/kg by intravenous route. The serum cefquinome concentrations in adult goats were determined up to 24 hrs after drug administration by using microbiological assay method. In the present study the therapeutic concentrations of cefquinome were maintained for more than 8 hours (0.08 ± 0.011mcg/ml) after intravenous administration and intramuscular administration and at the time of 10th hr sampling the cefquinome was not detected in serum after either intramuscular or intravenous administration.

Background and aim Folate is involved in a number of metabolic pathways. Red blood cell (RBC) folate is a well-established indicator of folate intake. However, studies focused on the association between RBC folate and coronary heart disease (CHD) are limited. The aim of the current study was to investigate the effect of RBC folate concentrations on the presence of CHD in a nationally representative sample of American adults. Methods and results In the 1999–2012 National Health and Nutrition Examination Survey (NHANES), 22,499 subjects aged 30–74 years with RBC folate concentrations, CHD status and responses to co-variates questions were included; 822 (3.65%) participants were identified as having CHD. Bio-Rad Quanta Phase II radioassay and microbiological assay were used to measure RBC folate concentrations. Firstly, we treated RBC folate as a categorical variable, based on RBC folate tertiles, and used logistic regression analysis to display the RBC folate and CHD relationship. Secondly, we explored associations using a combination of restricted cubic spline and logistic regression models, stratified by sex. After adjusting for several well-established traditional CHD risk factors, RBC folate was positively related to CHD presence in the total population and the association was more pronounced among males than females. A J-shaped pattern was observed in RBC folate concentrations for females. Conclusion Elevated RBC folate concentrations were associated with higher CHD risk. Further investigation is needed to test the association in large-scale follow-up studies.

A wide range of Indian foods (cereals, pulses, vegetables and milk based preparations) were analysed for five folate vitamers naturally present in the foods (n = 44). A liquid chromatography–mass spectrometry (LC–MS/MS) method using reversed phase chromatography and tandem mass spectrometry, coupled via positive mode electrospray ionization was used for the detection and quantification of the vitamers. The optimized LC–MS/MS method was capable of analysing the five most commonly-occurring folates (folic acid, 5-methyl tetrahydrofolic acid, tetrahydrofolic acid, 10-formyl folic acid and 5-formyl tetrahydrofolic acid) in 20 min. Quantification of folates was performed using 13C labelled internal standards. 5-methyl tetrahydrofolate was predominant in cereals, pulses and vegetable preparations. Fermented cereal preparations, beverages (coffee and tea) and green leafy vegetables were the main sources contributing to 5-formyl THF. Folic acid was identified in home-made yoghurt. All the values obtained in the present study using LC–MS/MS were compared to the total folate analysed using the microbiological assay in 2010 to generate data on the same foods. Findings suggest that the data obtained using both techniques showed agreement in the values (total folate calculated by adding the individual vitamers in the case of the LC–MS/MS values) particularly when foods were predominant in 5 methyl tetrahydrofolate.

Subinhibitory levels of antibiotics in reservoirs highly affected by anthropogenic activity, e.g., wastewater treatment plants, have a profound impact on the development and spread of resistant bacteria in the biosphere. As an aid, advanced oxidation processes (AOPs) have been suggested to eliminate the antibacterial activity of several antibiotics, and this activity has been followed using conventional antibiotic susceptibility tests. While the antibacterial activity can hardly be monitored below the minimum inhibitory concentration (MIC) with these assays, the selective pressure on a bacterial population might remain in this concentration range. To assess the applicability of an AOP for eliminating the subinhibitory effects of antibiotics on selection of resistant bacteria, a microbiological assay is introduced. The test is based on the dynamics of a mixed bacterial population in response to the presence of antibiotics in a concentration range well below the MIC in a synthetic wastewater matrix. Sensiti...

To investigate the often reported disagreement in food folate quantitation between the microbiological assay and high-performance liquid chromatography methods, different foods were analyzed both by a microbiological assay and by a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. For the LC-MS/MS analysis we emphasize the need for complete deconjugation of polyglutamic folate forms. Moreover, our results revealed no need for an additional enzyme treatment except in the deconjugation step. To check the efficiency of deconjugation without additional sample preparations, the amount of diglutamates was quantified and samples were screened for additional polyglutamates. A thorough investigation of a substance with a polyglutamate chain deconjugated like the folates revealed that it was an oxidation product of 5-methyltetrahydrofolate, a pyrazino-s-triazine called MeFox in previous reports. The latter is not microbiologically active and, therefore, does not contribute to the amount of total folates. But we found it is commonly present in foods, especially in those low in ascorbic acid. The microbiological assay showed different responses to the single vitamers. Therefore, it was necessary to perform calibration with the folate that had the highest portion in the folate distribution. The investigations showed that both methods can provide similar results when they both include a deconjugation step. This is particularly important for LC-MS/MS but probably also for the microbiological assay. Additionally, consideration of the folate distribution was found to be crucial for the accurate calibration of the microbiological assay.

This paper proposes a microbiological method in microtitre plates for the detection of residues of quinolones in milk. The method uses spores of Bacillus licheniformis in culture medium with a redox combination of indicators and gives a response time of 5.5 h. This method detects 92 μg L−1 of ciprofloxacin, 63 μg L−1 of danofloxacin, 109 μg L−1 of enrofloxacin, 101 μg L−1 of marbofloxacin and 109 μg L−1 of sarafloxacin in milk. Therefore, the assay is easy to develop and to use in laboratory, allowing analysis of large numbers of samples at low cost. Due to its good sensitivity to quinolones, this assay can be used as a complementary test of commercial microbiological methods and thereby improve food security.

Background: Ensuring the quality of infant and pediatric formulas and adult nutritionals is of utmost importance for the health and safety of rapidly urbanizing Indian population. B12 is an important water-soluble vitamin, which is fortified externally in such nutritional formulations. The Bureau of Indian Standards (BIS) has a recommended microbiological assay–based method for determination of vitamin B12 that is not precise and accurate enough to meet the label claim requirements of infant, adult, and/or pediatric nutritionals. The AOAC Official Method 2011.10 was originally developed under the AOAC Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN) for the determination of vitamin B12 in infant and pediatric formulas and adult nutritionals. However, those SPIFAN matrixes did not contain malt and other indigenous cereal and legume flour (with or without cocoa powder), which are commonly found in Indian formulations. Thus, there is a need to replace this method with a more precise and accurate method. Objective: This study was undertaken to validate the AOAC Official Method 2011.10 on vitamin B12 in ‘Indian’ infant and pediatric formulas and adult nutritionals. Methods: The single-laboratory validation (SLV) of AOAC Method 2011.10 was carried out as per the AOAC Guidelines in six Indian pediatric and adult nutritional formulas to verify its fitness for purpose. Cobalamin in the sample was converted to cyanocobalamin on treatment with potassium cyanide. The sample was then subjected to clean up through a C18 cartridge. Vitamin B12 in the eluted extract was separated from other components using size-exclusion column chromatography followed by a C18 column. The HPLC analysis was carried out at 550 nm. Results: Diastase treatment and C18 solid-phase extraction cleanup satisfactorily removed the matrix interference. The relative standard deviation of the determined values in 30 samples each from 6 selected Indian products and NIST SRM 1849a was

The aim of this work was to evaluate the effect of the addition of citrus fibers, from blood orange and lemon peels to produce a functional durum wheat bread. Breads fortified in fiber were packaged under a modified atmosphere (MAP) and stored at 25°C up to 120 days. No significant differences were observed with respect to the specific volume and weight, internal structure, pH and titratable acidity among the bread samples obtained using different types and percentages of fibers. Storage time, at 30 up to 90 days, affected significantly the bread firmness and caused significant differences in 5-hydroxymethylfurfural (HMF) levels in all bread samples. In fortified breads with citrus fibers the yeast and mold counts showed values of approximately 1 log10 cfu/g for the first 30 days and 3.5 log10 cfu/g at the end of storage. The results of the sensory evaluation highlight that loaves enriched with blood orange and lemon fibers showed a citrus flavor but had a similar overall evaluation respect to control bread produced without addition of citrus fiber. The results of this study showed that the addition up to 2% of blood orange and lemon fibers in wheat whole durum flour is a possible strategy to produce "high fibre" bread.

Bull’s eye rot (BER) is a major economic postharvest disease of apple and pear that can be caused by four Neofabraea species i.e., N. perennans, N. alba, N. malicorticis and N. kienholzii. In the growing regions of Central Washington, BER is predominantly caused by N. perennans. The fungus infects fruit preharvest, and because of the dry growing season in the PNW, infections remain latent with symptoms expressed only after 3-4 months of storage, when incidences as high as 20% can been seen in rainy seasons, especially on susceptible cultivars. To detect the infections more efficiently and quickly before symptoms become visible at point-of-care locations, a portable diagnostic tool termed LAMP, Loop mediated isothermal amplification, was developed for early detection of N. perennans. The LAMP technique was optimized and tested for specificity and sensitivity using DNA extracted from pure culture and was used to analyze apple fruits inoculated with N. perennans at different concentrations and sampling times pre- and postharvest. Results indicate that LAMP detected DNA of N. perennans at concentrations as low as 0.001 ng/[mu]l after only 10 minutes of running the reaction. Further validation of the LAMP assay using a portable thermocycler (Genie II®), indicated that LAMP can detect the fungus on apples inoculated with spore suspensions at concentrations of 103 spores/ml or higher prior to commercial maturity without the need for DNA extraction, as well as detecting variability between samples. The assay was further validated using commercial samples of Pinova apples from organic and conventional orchards, demonstrating the ability of this technique to amplify N. perennans from asymptomatic fruit in a commercial setting. LAMP has potential to be used at point of care facilities to better inform the management of BER as fruit goes from harvest into storage, and it has potential to be utilized to better understand the life-cycle of the pathogen.

The aim of this study was to determine the prevalence of quality herbal products sold in Thailand. The quality of herbal products focuses on three specific issues: i) the legality of the labelling on herbal products, ii) microbial contamination and iii) corticosteroid adulteration in herbal products. This cross-sectional study randomly selected 431 herbal products bought from online stores, markets, grocery stores, department stores, traditional practitioners, pharmacies, and hospitals across Thailand, in 2016. The names of the selected products were guarded by a double - blind barrier when testing for microbial limitation and corticosteroids; a condition that was extended to the data analysis process. The microbiological assay was based upon total aerobic microbial counts and total combined yeasts/moulds counts. Corticosteroid tests were performed using the DMSc Steroid Test Kit produced by the Department of Medical Sciences, Thailand, and thin layer chromatography. The prevalence of quality herbal products in Thailand was 14.5%. Forty-four percent of the examined products were registered with the Thai FDA. Seventy percent of the products were contaminated with aerobic bacteria and 11% with combined yeasts/moulds. Less than one percent was adulterated with prednisolone. Therefore, it can be concluded that the Thai FDA urgently needs to enforce “Good Manufacturing Practices’ (GMPs), and devise a process of post-marketing surveillance of the quality of herbal products’.

The used of Gentamicin is still regarded for overcoming infectious diseases. Several preparations of the gentamicin are available in the market as injection, eye or ear drops, and topical dosage forms. Based on the standard method, determination of the gentamicin is carried out by microbiological assay. Several instrumental method have been reported. In this research, Thin Layer Chromatography-Densitometry (TLC-Densitometry) has been validated and used for determining gentamicin in cream dosage form. The TLC was carried out on a Silica gel GF254 using KH2PO4 20% and ninhidrin 2% solution (in ethanol 96%) as eluent and spots visualization respectively. Three spots appeared on the chromatogram having Rf value 0.51, 0.47, and 0.36. All the spots gave maximum absorption at 400 nm. The spot with Rf value of 0.51 was the highest intensity. Limit of Detection (LoD) and Limit of quantitation (LoQ) of the major component was 0.019 and 0.064 respectively. The other validation characteristics met the requirement for determination of gentamicin in the cream dosage form.

Antibiotics are the chemotherapeutic agents that kill or inhibit the pathogenic microorganisms. Resistance of microorganism to antibiotics is a growing problem around the world due to indiscriminate and irrational use of antibiotics. In order to overcome the resistance problem and to safely use antibiotics, the correct measurement of potency and bioactivity of antibiotics is essential. Microbiological assay and high performance liquid chromatography (HPLC) method are used to quantify the potency of antibiotics. HPLC method is commonly used for the quantification of potency of antibiotics, but unable to determine the bioactivity; whereas microbiological assay estimates both potency and bioactivity of antibiotics. Additionally, bioassay is used to estimate the effective dose against antibiotic resistant microbes. Simultaneously, microbiological assay addresses the several parameters such as minimal inhibitory concentration (MIC), minimum bactericidal concentration (MBC), mutation prevention concentration (MPC) and critical concentration (Ccr) which are used to describe the potency in a more informative way. Microbiological assay is a simple, sensitive, precise and cost effective method which gives reproducible results similar to HPLC. However, the HPLC cannot be a complete substitute for microbiological assay and both methods have their own significance to obtain more realistic and precise results.

Basil is one of a plant that has antibacterial activity on Staphylococcus aureus , the cause of furuncle . To cure the desease it is require preparations with a good penetrability, i.e gels and ointments . The goal of the research was to obtained the most stable both gel and ointment formulations of basil seed oil ( Ocimum basilicum Linn.) pharmaceutically and effective microbiologically. Physical stability evaluations were performed by the parameters were organoleptic, viscosity measurement, homogeneity, spreadability, and flowing type at 5 o C and 35 o C for 12 hours on each temperature for 10 cycles. Organoleptic assay showed that there was no change in color, odor and concistency of all formula. Based on statistic analyses by Randomized design exhibited a significant change of gel and ointment formulation during storage. Homogeneity test showed that all formula possess even particle distribution. Based on rheogram exhibited thixotrophy-plastic flow type. Microbiological assay carried out by observe the formed inhibitory zone. Physical evaluation obtained formula II of gel was the most stable (12% basil seed oil) and formula III of ointment (18% basil seed oil) and microbiologically effective was formula III. While the most effective microbiologically in inhibit the growth of Staphylococcus aureus was gel preparation in 18% concentration of basil seed oil .

Folic acid (FA) is an important vitamin for human growth and development, especially for pregnant women. A sensitive, rapid, and accurate FA detection method is required to assess the nutritional quality and safety of foods. A monoclonal antibody against FA was prepared for the development of indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) and lateral-flow immunochromatographic assay (ICA) strip. The 50% inhibitory concentration and limit of detection of ic-ELISA were 0.12 and 0.018 ng/ml, respectively. The visual limit of detection and cut-off values of the lateral-flow ICA strip were 0.5 and 2.5 ng/ml, respectively. Using the ICA strip, FA recovery rates were 89–98% from energy drinks and 73–87% for milk samples and were in good agreement with those obtained from the conventional microbiological assay method. Our developed methods are sensitive, convenient, effective, and suitable for on-site detection and rapid mass screening of food samples.

Cefquinome is one of the fourth generations of cephalosporins developed for veterinary use in treatment of respiratory diseases that are considered the second causes of death in calves. Non-steroidal anti-inflammatory drugs such as flunixin (NSAIDs) are widely prescribed with antibacterial agents in multiple drug prescriptions. The present study aimed to investigate the effect of co-administration of flunixin on the disposition kinetics of cefquinome after intramuscular injection in 10 diseased calves (Pasturella heamolytica infected). Cefquinome was injected in a single dose (2 mg/kg BW) in 5 diseased calves alone and coupled with flunixin (1mg/kg BW) in the other 5 diseased calves. Blood samples (5 mL) were collected from the right jugular vein of each calve immediately before treatment and at intervals of 0, 5, 10, 15 and 30 min, 1, 2, 4, 6, 8, 12, and 24 hours, 48, 72, 96 and 120 h (5 days) after cefquinome administration. The obtained samples were assayed with the plate microbiological assay method using Sarcina lutea (ATCC 9341) as test organism. The plasma cefquinome concentration at 5 min after intramuscular injection of cefquinome alone and coupled with flunixin was 0.27 ± 0.05 μg/mL and 0.35 ± 0.12 μg/mL, respectively and reached the highest concentration (1.02 ± 0.12 μg/mL and 1.02 ± 0.08 μg/mL) at 1 h, respectively. The obtained data showed no significant effect of coupled administration of flunixin with cefquinome on either concentration or peak concentration of cefquinome in plasma of diseased calves. It is concluded that flunixin can be used successfully with cefquinome in treatment of bacterial respiratory diseases associated with inflammation in calves.

Ready-to-eat breakfast cereals have been voluntarily fortified with folic acid since 1995, with the purpose of reducing the prevalence of neural tube defects in utero. Using data from the recent Australian Health Survey, this study aimed to estimate folate intake from one serving of breakfast cereals (median amount). Various commercial brands were purchased in 2002 (n = 19) and in 2014 (n = 14); folate was determined by microbiological assay and high-pressure liquid chromatography (HPLC). Total folate (μg/100 g) in 2002 and 2014 selections were 144-633 and 147-564, respectively, and mostly comparable to nutrition labels. Folic acid (2014 selection) using HPLC, ranged from 85 to 411 μg/100 g. Intake of 51 g cereals/serving by individuals ≥ 2 years could contribute 75-288 μg dietary folate equivalent. It seems that folic acid intake among children (2-3 years) exceeds the recommended dietary intake, when certain brands of breakfast cereals are consumed. Accordingly, the benefits and potential detrimental effects of the voluntary fortification need to be further explored.