Yeast Identification

In the last century, the exponential increase of industrial food production led to the disappearance of "Italian traditional niche products". However, national regulations allowed the preservation of several of these products, including the burrata cheese. Twenty-one samples from three different batches of "Burrata di Andria" Protected Geographical Indication (PGI) were purchased from dairy factories of the PGI consortium. Moisture value of PGI Burrata cheese was significantly higher than that before the PGI release. Moreover, a significantly lower NaCl value was detected in PGI raw milk Burrata cheeses with respect to non-PGI ones, while an opposite situation was detected in pasteurized milk Burrata cheeses. As for pH, in all PGI products lower values were observed with respect to non-PGI products, which resulted significant only in pasteurized ones. No Salmonella spp., Listeria monocytogenes, and Bacillus cereus were detected, while nine samples were positive for a nonpathogenic strain of Yersinia enterocolitica. Total viable count (TVC) and Escherichia coli resulted significantly lower in pasteurized than in raw milk PGI Burrata cheese samples. Although samples analyzed can be considered microbiologically safe, these were borderline and/or unsatisfactory for E. Coli and coagulase-positive staphylococci (CPS) according to process hygiene criteria established by European regulation. Therefore, different strategies should be adopted to improve products hygiene in the considered dairy factories.

In the last century, the exponential increase of industrial food production led to the disappearance of “Italian traditional niche products”. However, national regulations allowed the preservation of several of these products, including the burrata cheese. Twenty-one samples from three different batches of “Burrata di Andria” Protected Geographical Indication (PGI) were purchased from dairy factories of the PGI consortium. Moisture value of PGI Burrata cheese was significantly higher than that before the PGI release. Moreover, a significantly lower NaCl value was detected in PGI raw milk Burrata cheeses with respect to non-PGI ones, while an opposite situation was detected in pasteurized milk Burrata cheeses. As for pH, in all PGI products lower values were observed with respect to non-PGI products, which resulted significant only in pasteurized ones. No Salmonella spp., Listeria monocytogenes, and Bacillus cereus were detected, while nine samples were positive for a nonpathogenic s...

The quality evaluation of white soft cheese in the Egyptian markets is an urgent need to create awareness among population about the existing situation and to protect the consumer's health and rights. Thus, the aim of this work was to determine AFM levels and evaluate microbiological properties in 1 commercially available white soft cheese collected from local markets as well as to evaluate the manufactured control samples using the Hazard Analysis Critical Control Points (HACCP) system. Results revealed that AFM1 concentration was detected in almost half of the samples (46.66%) with an overall mean of 14.39 ng/kg and was detected in 24 samples at levels below 1.0 ng/kg and in 15 samples at levels ranging from 10 to 50 ng/kg, whereas about 3 samples were higher than 50 ng/kg. All positive samples of white cheeses exceeded the Egyptian regulations (free from AFM ). The microbiological analysis revealed that all samples contained high 1

A case study of the indigenous technology for processing white soft cheese was conducted in the town of Lumban in Laguna, Philippines. Information was gathered on the demographic profile of two cheese makers; their indigenous knowledge and practices; and the nature, volume, and marketing system of the kesong puti business. The receptivity of the cheese makers to the DTRI technology on white soft cheese processing and the chemical and microbiological qualities of the kesong puti produced in the town were determined. The cheese making business of the two cheese makers' families dates back to 1900 and 1928, respectively. Cheese making is their main occupation. Both cheese makers use bahay asim (abomasum) to coagulate the milk.The average income from the business ranged from PhP15,000-40,000/month. Only one cheese maker expressed interest in the demonstration of the DTRI method for making white soft cheese. A major constraint to the adoption of the DTRI technology according to the cheese maker was the longer processing time and lower cheese yield obtained. The other cheese maker had confidence in her own traditional cheese making technology and expressed no interest in the DTRI method. Lumban cheese had 65.96-67.27% moisture, 2.20-2.60% ash, 11.49-13.14% total protein, 10.17-10.33% fat and 2.03-2.15% salt. The pH ranged from 6.18 to 6.34. The total bacterial counts and yeast and mold counts ranged from 1.4-8.4 x 10 6 colony forming units (cfu)/g and 1.7-3.4 x 10 4 cfu/g, respectively. The coliform colonies were too numerous to count at 10 -1 dilution, which is beyond the 10 cfu/g coliform count limit set by the Philippine Bureau of Food and Drugs.

This research was conducted to study the morphological characteristics of the main yeast strains isolated from table olives (colour, shape of colonies, production of compounds around the colonies) and the genetic ones (molecular characterization, two types of phylogenetic tree).
In particular, smears were made on Petri dishes, using two different types of agar, differential, and non-differential, belonging in total to 23 yeast strains (identified/unidentified). The microbial cultures of origin from which the smears were made, belong to the species: Wickerhamomyces anomalous, Candida diddensiae, Metschnikowia pulcherrima, Pichia membranifaciens.
After making smears on Petri dishes of the twenty tree strains of the olive’s yeasts, the colonies were grown at an incubation temperature of 25°C for 48 hours. The morphological characteristics of the yeasts were observed, such as the size of the colonies (microns), the shape, the colour, the formation of particular compounds produced accumulated around the same colonies and visible at a first visual examination.
These species of yeasts belonging to 23 strains, were subsequently identified with three molecular identification techniques, one less specific and the other two more accurate. In particular, a RAPD-PCR was used, which had the aim of identifying the cultivated yeast strains that are not present in the database, therefore of which there was no certainty of the genus and species. A phylogenetic tree, built with the RAPD-PCR profiles that allowed to group the yeast strains into six groups and to identify three strains of C. diddensiae species with a very high percentage of similarity of the 95,3%, therefore very similar to each other at the genetic level. Furthermore, the phylogenetic tree validated the results of the RAPD-PCR for three strains of C. diddensiae.
Finally, the last molecular identification technique, more accurate than the previous ones, was the phylogenetic tree with blast sequences. Through the use of these technique, ‘specific DNA sequences’ called ‘blast sequences ‘, that precisely indicate the genus and the species of certain yeast strains (high percentage of genetic similarity) were compared with the twenty-three yeast strains of the study (identified and unidentified).
In this way, it was possible to identify yeast strains that are not present in the database with certainty of the genus and species and to confirm and strengthen the initial results of molecular analysis (RAPD-PCR) with a higher percentage of genetic similarity for the genus and species of the study strains already present in the database and also validate the initial results of molecular identification (RAPD-PCR) for the study strains do not present in the database.

The quality evaluation of white soft cheese in the Egyptian markets is an urgent need to create awareness among population about the existing situation and to protect the consumer's health and rights. Thus, the aim of this work was to determine AFM levels and evaluate microbiological properties in 1 commercially available white soft cheese collected from local markets as well as to evaluate the manufactured control samples using the Hazard Analysis Critical Control Points (HACCP) system. Results revealed that AFM1 concentration was detected in almost half of the samples (46.66%) with an overall mean of 14.39 ng/kg and was detected in 24 samples at levels below 1.0 ng/kg and in 15 samples at levels ranging from 10 to 50 ng/kg, whereas about 3 samples were higher than 50 ng/kg. All positive samples of white cheeses exceeded the Egyptian regulations (free from AFM ). The microbiological analysis revealed that all samples contained high 1

The yeasts present during the ripening process of ewes' and goats' cheeses produced in a small traditional dairy in Mediterranean Spain were isolated and identified. Five hundred and thirty strains pertaining to eleven yeast species representing eight genera were identified using molecular methods. Debaryomyces hansenii was the yeast species most frequently isolated in all cheeses. Other yeast species commonly found in dairy products were present at the first maturing weeks. Two yeast species Trichosporon coremiiforme and Trichosporon domesticum have been reported in cheeses for the first time, and Meyerozyma guilliermondii has been newly isolated from goats' cheeses. Yeast species composition changed greatly along the process; although, D. hansenii dominated at the end of ripening in all cheeses. Most yeast isolates were able to hydrolyze casein and fatty acid esters. One hundred and eighty seven D. hansenii isolates were genotyped by PCR amplification of M13 satellites and a UPGMA dendrogram was constructed. The majority of isolates were grouped in 5 clusters while 7 profiles were represented by 1 to 3 isolates. These results demonstrate the genetic heterogeneity present in D. hansenii strains isolated from raw milk cheeses.

Six types of unripened raw milk fresh (Robiola, Crescenza, Primo sale and Formaggella) and "pasta filata" cheeses (Mozzarella and Burrata) were evaluated for microbiological parameters. No Listeria monocytogenes or Salmonella spp. were detected, but high microbial counts were revealed. Significantly higher Total Viable Counts (TVC) (P=0.002) and Enterobacteriaceae counts (P<0.001) were observed in "fresh cheese" than in "pasta filata" samples. Values > 6 Log CFU/g were found in 81.3% of fresh vs 50% in pasta filata for TVC and 65.6% vs 12.5% for Enterobacteriaceae, respectively. An evident contamination by Escherichia coli, Coagulasepositive Staphylococci and Pseudomonas spp. was detected in all the cheeses: the causes could be the improper hygiene of the artisanal production practices and the permanence of the cheeses on the refrigerated shelves. A careful attention to the respect of the good manufacturing practices is suggested to avoid the presence of initial high bacterial loads.

Ten strains of a new endophytic ascosporeforming, methanol-assimilating yeast were isolated from the galls induced by sawflies on the leaves of willows in the Losiny Ostrov National Park (Moscow region). Standard phenotypical tests and phylogenetic analyses of 18S rRNA gene, 5.8S-ITS gene region and 26S rRNA gene (D1/D2 domains) sequences showed that the species belongs to the genus Ogataea. We describe it as Ogataea cecidiorum and designate type culture KBP Y-3846 (= CBS 11522 T = VKM Y-2982 T = VKPM Y-3482 T = MUCL 52544 T = NCAIM Y.01965 T) as the type strain. The new species was registered in MycoBank under MB 515233.

Scolytine bark beetles are the most destructive pests of conifers; they sometimes aggregate in such large numbers that they actually kill their hosts. They maintain close relationships with yeasts and fungi, in particular those that are assumed to aid in digestive, detoxification processes and pheromone production. In this study, 403 yeast strains were isolated from the guts, ovaries, eggs and frass of nine bark beetle species in the genus Dendroctonus Erichson. The beetles were collected from 10 conifer species at 34 locations in Mexico, Guatemala and the USA. Yeast identification was based on partial DNA sequences from 18S rDNA, 26S rDNA and internal transcribed spacer (ITS1), as well as morphological and physiological characteristics. A combined phylogenetic analysis delimited 11 clades with sequences similar to Candida arabinofermentans, C. ernobii, C. membranifaciens (including C. lessepsii, Pichia mexicana and P. scolyti), C. oregonensis, C. piceae, Kuraishia capsulata (including K. capsulata and K. cf. molischiana), Pichia americana, P. canadensis, P. glucozyma, P. guilliermondii and an undescribed species of Candida. Nucleotide divergences between the major clades were at least 5% while, with the exception of 30 isolates, yeasts within clades differed from named reference species at fewer than 1% of the nucleotide sites. There do not appear to be obligate relationships between particular yeasts and specific anatomical partitions, nor between particular yeasts and bark beetle species. Some yeasts do appear to be preferentially associated with bark beetles feeding on different conifer genera and therefore host plant defences may limit yeast community diversity in Dendroctonus.

The aim of this study was to assess the microbiological quality and safety of some cheeses produced in Apulia region, Southern Italy, and classified as "traditional agri-food products." A total of 120 samples were collected and analyzed. From results, 82.5% of cheeses analyzed, had a satisfactory microbiological quality according to the European legislation. The coagulase positive Staphylococci (CPS) count was above the limits set by the European legislation in 17.5% of the samples analyzed. Within the 20 strains of Staphylococcus aureus (S. aureus) isolated, 12 were positive for the detection of the enterotoxin genes. Furthermore, nine strains of S. aureus were positive to reverse passive latex agglutination (SET-RPLA) assay. One of the two samples of Cacioricotta di Vacca Fresco, with a CPS load of 3.02 × 10 5 cfu/g, showed unsatisfactory microbiological safety because was positive to SET-RPLA. Based on the findings, it is essential to apply and maintain good hygiene practices throughout the food chain, especially for traditional agri-food products, which are often produced in small family processing plants using production protocols based on the experience handed down from father to son. Practical applications The traditional agri-food products are foodstuffs much appreciated by consumers because of their organoleptic qualities and taste that brings back to the genuine flavors of the past. However, the hygienic quality and safety of these products is linked to the personal experience of food business operator and not to the regular checking of microbiological parameters that validate the acceptability of foodstuffs and their manufacturing, handling and distribution processes. The results of this study can contribute to the risk analysis for these particular products, and underline the need to apply and maintain good hygiene practices throughout the production chain.

A total number of 60 samples from traditional fermented milk “Rayeb” was randomly collected from different localities in Sfax governorate, Tunisia and subjected to determination of pH, sensory evaluation and determination of total yeast and mold count, Coliforms count, total staphylococcal count, Staphylococcus aureus count and detection of Salmonella. The obtained results revealed that pH was ranged from 3.8 to 4.7, 40 out of 60 samples (66.67 %) had the grade good in sensory evaluation, the mean yeast and mold count was 1.34×107± 2262054 CFU/mL, the mean coliforms count was 3.707×103 ± 1510 CFU/mL. the mean count of total staphylococci was 5.10×103 ± 1257 CFU/mL, the mean S. aureus count was 68.4 ± 13 CFU/ mL. The incidence of pathogens in fermented foods, as highlighted in this study suggests that measures that minimize the risk of food-borne illnesses should be taken. The application of Hazard Analysis and Critical Control Point (HACCP) as advocated by the WHO should be applied ...

Multi Fragment Melting Analysis System (MFMAS) is a novel approach that was developed for the species-level identification of microorganisms. It is a software-assisted system that performs concurrent melting analysis of 8 different DNA fragments to obtain a fingerprint of each strain analyzed. The identification is performed according to the comparison of these fingerprints with the fingerprints of known yeast species recorded in a database to obtain the best possible match. In this study, applicability of the yeast version of the MFMAS (MFMAS-yeast) was evaluated for the identification of food-associated yeast species. For this purpose, in this study, a total of 145 yeast strains originated from foods and beverages and 19 standard yeast strains were tested. The DNAs isolated from these yeast strains were analyzed by the MFMAS, and their species were successfully identified with a similarity rate of 95% or higher. It was shown that the strains belonged to 43 different yeast species ...

Multi Fragment Melting Analysis System (MFMAS) is a novel approach that was developed for the species-level identification of microorganisms. It is a software-assisted system that performs concurrent melting analysis of 8 different DNA fragments to obtain a fingerprint of each strain analyzed. The identification is performed according to the comparison of these fingerprints with the fingerprints of known yeast species recorded in a database to obtain the best possible match. In this study, applicability of the yeast version of the MFMAS (MFMAS-yeast) was evaluated for the identification of foodassociated yeast species. For this purpose, in this study, a total of 145 yeast strains originated from foods and beverages and 19 standard yeast strains were tested. The DNAs isolated from these yeast strains were analyzed by the MFMAS, and their species were successfully identified with a similarity rate of 95% or higher. It was shown that the strains belonged to 43 different yeast species that are widely found in the foods. A clear discrimination was also observed in the phylogenetically related species. In conclusion, it might be suggested that the MFMAS-yeast seems to be a highly promising approach for a rapid, accurate, and one-step identification of the yeasts isolated from food products and/or their processing environments.

Background and Aim: Milk is a food of high nutritional value, which occupies an undeniable place in the human food ration, but is an ideal medium for microbial growth. This study aims to assess the hygienic quality of local raw and fermented milk from the Liptako-Gourma region in Niger. Materials and Methods: We performed physical and bacteriological analyses on 330 samples of bovine milk from local breeds, including 110 individual milk samples (per cow), 110 fermented milk samples, and 110 blended milk samples. The microbiological parameters were determined using standard methods. Results: The physical analysis revealed temperatures during sample collection for all milk types between 35.2°C and 37.8°C. The average pH of fermented milk varied between 3.16 and 4.92 and those of individual and blended raw milks between 5.42 and 6.98. The titratable acidity varied from 15° to 18.1°D for raw milk and between 59° and 122°D for fermented milk. The average density of individual and blended...

The cheese microbial ecosystem is complex, and the presence of nonstarter adventitious microorganisms in milk may have an influence on the organoleptic characteristics of cheese. The aim of this study was to analyze the composition and diversity of the fungal flora of raw milk destined for cheesemaking from 19 dairy farms in Quebec and to monitor their evolution throughout ripening. Six hundred ten yeast and mold isolates were collected from raw milk and raw milk cheeses over a 9-month period. Based on the sequences of the rDNA ITS1-5.8S-ITS2 region, 67% of the raw milk isolates were yeasts, which were assigned to 37 species across 11 genera, while 33% were molds, which were assigned to 33 species across 25 genera. A semi-quantitative analysis of the yeasts and molds in the raw milk from four farms was performed over a 5-month period. The composition and diversity of the fungal microflora were totally different for each farm, each of which had a unique species profile. To determine whether adventitious yeast strains from the milk could develop in raw milk cheese, a multilocus-sequence-typing (MLST) analysis was performed on 13 Issatchenkia orientalis (syn. Pichia kudriavzevii, anamorph: Candida krusei) isolates. The same MLST genotypes were identified for strains independently isolated from raw milk and raw milk cheese from a farm processing its own milk. This study contributes to

Background and Aims: Non-steroidal anti-inflammatory drugs (NSAIDs) are consumed in treatment of inflammation and pain in many conditions. New derivatives of Aryliliden-2 (-3 chloro anilino) nicotinic acid hydrazide, were synthesized in order to be evaluated for anti-inflammatory and analgesic effects in vivo. New types synthesized based upon modulation the NSAIDs side effects as well as improving their safety profile. Methods: Anti-nociceptive activity was assessed by acetic acid induced writhing response. The suspention test compounds administration (i.p.) intraperitoneally (1% v/v; 10 mg/kg), then 30 min after aqueous acetic acid solution (1% v/v; 10 mg/kg) was administered intraperitoneally, analgesic activity was recorded via counting the number of writhes during 30 min after acetic acid injection in mice,then analgesic activity was calculated using the following formula, % inhibition = [(control mean-test mean) / control mean ] ×100 Anti-inflammatory activity was determined using carrageenan-induced edema test in the hindpaws of rats,carrageenan (0.1 mL of 1% w/v in saline) was injected subcutaneously in the rat footpads 1 h after i.p. administration of compounds. The increase in paw volume (0.5, 1, 2, 3 and 4 hours after carageenan) is calculated as percentage compared with the basal volume. Results: Among all of analogues, 4-pyr, 3-F and 3-(Ome) with (90%, 57% and 60% inhibition) had the most potent analgesic activity and the compounds 4-pyr, 3-Cl, 2,4(Ome)2, 3-F and 3-(Ome) exhibited the most potent anti-inflammatory activity, (investigations showed no ulcerogenic effects). Conclusions: We demonstrated that most of The derivatives synthesized of arylidene-2-(-3chloro anilino) nicotinic acid hydrazides were able to reduce the AcOH induced constrictions in the mice and also exhibited anti-inflammatory activity ranging from moderate to good in comparison with Carboxymethyl cellulose (CMC) and Niflumic acid as the reference drugs.

All the reports to date on the anti-inflammatory activity of chitooligosaccharides (COS) are mostly based on in vitro methods. In this work, the anti-inflammatory activity of two COS mixtures is characterized in vivo (using balb/c mice), following the carrageenaninduced paw edema method. This is a widely accepted animal model of acute inflammation to evaluate the anti-inflammatory effect of drugs. Our data suggest that COS possess anti-inflammatory activity, which is dependent on dose and, at higher doses, also on the molecular weight. A single dose of 500 mg/kg b.w. weight may be suitable to treat acute inflammation cases; however, further studies are needed to ascertain the effect upon longer inflammation periods as well as studies upon the bioavailability of these compounds.

Food contamination, either from microbiological or chemical, is the highest concern for public health. The dairy products such as cheese have an essential role in human nutrition but have been linked with outbreaks of foodborne illness. The cheese production process contains the steps that might be the reasons of bacterial pathogens contamination. The primarily purpose of this study was to detect the microbiological quality and Lactic Acid flora of cheeses at retail sale in bazaars of Bayramiç, Ezine and Kepez (towns of Çanakkale) which were produced at home or small dairies. The presence of Total Mesophilic Aerobic Bacteria (TMAB), Total Psychrophilic Aerobic Bacteria (TPAB), yeast and moulds, coliform bacteria, coagulase positive Staphylococcus spp., and Escherichia coli O157:H7 (EHEC) were investigated. The mean TMAB counts were assessed as 3.63 × 10 8 cfu/g (2.00 × 10 6-≥ 1.00 × 10 9 cfu/g), 2.19 × 10 8 cfu/g (1.00 × 10 4-≥ 1.00 × 10 9 cfu/g), and 1.55 × 10 8 cfu/g (2.00 × 10 6-≥ 1.00 × 10 9 cfu/g) in cheese samples from Bayramic, Ezine and Kepez, respectively. TPAB was not determinated. The mean counts of coliforms were determinated as 1.44 × 10 5 cfu/g (ND-≥ 1.00 × 10 6 cfu/g), 1.04 × 10 5 cfu/g (ND-≥ 8.00 × 10 5 cfu/g) and 5.71 × 10 1 cfu/g (ND-≥ 8.00 × 10 2 cfu/g) in cheese samples from Bayramiç, Ezine and Kepez respectively. E.coli 0157:H7 and coagulase positive Staphylococcus aureus were not detected. The average colony counts of Lactococcus spp., Enterococcus spp. and Lactobacillus spp. were detected as 2.07 × 10 8 cfu/g (ND-≥ 1.00 × 10 9 cfu/g), 2.43 × 10 6 cfu/g (ND-≥ 3.30 × 10 7 cfu/g) and 1.54 × 10 8 cfu/g (ND-≥ 1.00 × 10 9 cfu/g) in town Bayramiç; 1.86 × 10 8 cfu/g (ND-1.00 × 10 9 cfu/g), 1.46 × 10 5 cfu/g (ND-6.1 × 10 6 cfu/g) and 1.31 × 10 8 cfu/g (ND-≥ 1.00 × 10 9 cfu/g) in town Ezine; 9.57 × 10 7 cfu/g (ND-≥ 1.00 × 10 9 cfu/g), 1.23 × 10 5 cfu/g (ND-1.00 × 10 6 cfu/g) and 8.24 × 10 7 cfu/g (ND-≥ 1.00 × 10 9 cfu/g) in town Kepez, respectively. According to this study results, cheese samples produced without uncontrolled conditions such as in small dairy farms or home and sold in bazaars of Çanakkale have high microorganism loads and do not meet the microbiological criteria in Turkish Food Codex regulations.

A yeast strain was isolated during a study on vineyard-associated yeast strains from Apulia in Southern Italy. ITS and LSU D1/D2 rDNA sequences showed this strain not to belong to any known species and was described as the type strain of , a close relative of . Several secondary peaks appeared in the sequences, suggesting internal heterogeneity among the copies of the rDNA. This hypothesis was tested by sequencing single clones of the marker region. The analyses showed different levels of variability throughout the operon with differences between the rRNA encoding genes and the internally transcribed regions. and share high frequency variants, i.e., variants frequently found in many clones, whereas there is a large variability of the low frequency polymorphisms, suggesting that the mechanism of homogenization is more active with the former than with the latter type of variation. These findings indicate that low frequency variants are detected in Sanger sequencing as secondary peaks ...

Multi Fragment Melting Analysis System (MFMAS) is a novel approach that was developed for the species-level identification of microorganisms. It is a software-assisted system that performs concurrent melting analysis of 8 different DNA fragments to obtain a fingerprint of each strain analyzed. The identification is performed according to the comparison of these fingerprints with the fingerprints of known yeast species recorded in a database to obtain the best possible match. In this study, applicability of the yeast version of the MFMAS (MFMAS-yeast) was evaluated for the identification of food-associated yeast species. For this purpose, in this study, a total of 145 yeast strains originated from foods and beverages and 19 standard yeast strains were tested. The DNAs isolated from these yeast strains were analyzed by the MFMAS, and their species were successfully identified with a similarity rate of 95% or higher. It was shown that the strains belonged to 43 different yeast species ...

The aim of this study was to assess the microbiological quality and safety of some cheeses produced in Apulia region, Southern Italy, and classified as "traditional agri-food products." A total of 120 samples were collected and analyzed. From results, 82.5% of cheeses analyzed, had a satisfactory microbiological quality according to the European legislation. The coagulase positive Staphylococci (CPS) count was above the limits set by the European legislation in 17.5% of the samples analyzed. Within the 20 strains of Staphylococcus aureus (S. aureus) isolated, 12 were positive for the detection of the enterotoxin genes. Furthermore, nine strains of S. aureus were positive to reverse passive latex agglutination (SET-RPLA) assay. One of the two samples of Cacioricotta di Vacca Fresco, with a CPS load of 3.02 × 10 5 cfu/g, showed unsatisfactory microbiological safety because was positive to SET-RPLA. Based on the findings, it is essential to apply and maintain good hygiene practices throughout the food chain, especially for traditional agri-food products, which are often produced in small family processing plants using production protocols based on the experience handed down from father to son. Practical applications The traditional agri-food products are foodstuffs much appreciated by consumers because of their organoleptic qualities and taste that brings back to the genuine flavors of the past. However, the hygienic quality and safety of these products is linked to the personal experience of food business operator and not to the regular checking of microbiological parameters that validate the acceptability of foodstuffs and their manufacturing, handling and distribution processes. The results of this study can contribute to the risk analysis for these particular products, and underline the need to apply and maintain good hygiene practices throughout the production chain.

Scolytine bark beetles are the most destructive pests of conifers; they sometimes aggregate in such large numbers that they actually kill their hosts. They maintain close relationships with yeasts and fungi, in particular those that are assumed to aid in digestive, detoxification processes and pheromone production. In this study, 403 yeast strains were isolated from the guts, ovaries, eggs and frass of nine bark beetle species in the genus Dendroctonus Erichson. The beetles were collected from 10 conifer species at 34 locations in Mexico, Guatemala and the USA. Yeast identification was based on partial DNA sequences from 18S rDNA, 26S rDNA and internal transcribed spacer (ITS1), as well as morphological and physiological characteristics. A combined phylogenetic analysis delimited 11 clades with sequences similar to Candida arabinofermentans, C. ernobii, C. membranifaciens (including C. lessepsii, Pichia mexicana and P. scolyti), C. oregonensis, C. piceae, Kuraishia capsulata (including K. capsulata and K. cf. molischiana), Pichia americana, P. canadensis, P. glucozyma, P. guilliermondii and an undescribed species of Candida. Nucleotide divergences between the major clades were at least 5% while, with the exception of 30 isolates, yeasts within clades differed from named reference species at fewer than 1% of the nucleotide sites. There do not appear to be obligate relationships between particular yeasts and specific anatomical partitions, nor between particular yeasts and bark beetle species. Some yeasts do appear to be preferentially associated with bark beetles feeding on different conifer genera and therefore host plant defences may limit yeast community diversity in Dendroctonus.

2-Pyrazolines 7-12 bearing a carboxylic acid ester or a carboxamide side-chain have been prepared by treatment of the appropriate chalcone derivatives 2-6 with hydrazine hydrate or phenylhydrazine in hot acetic acid. 1-(2-Carboxyphenyl)-2-pyrazolines 24-30 and 1-(4carboxyphenyl)-2-pyrazolines 31-41 were synthesized by the reaction of chalcones with (2carboxyphenyl)hydrazine and (4-carboxyphenyl)hydrazine in hot acetic acid. Structures of all new compounds have been elucidated by microanalyses, 1 H-, 13 C-NMR and IR spectroscopic measurements.

Teleomorphic species of the genus Zygozyma and anamorphic species of the genus Myxozyma were examined phylogenetically. In phylogenetic trees based on 18S rRNA-, 26S rRNA-, mitochondrial small subunit rRNA-and EF-1α-gene sequences and concatenated sequences of the latter four regions, derived from the neighbor-joining method, the four species of the genus Zygozyma constituted four clusters, respectively, with low bootstrap values, indicating that all four species can be distinguished from one another at the generic level. The name of Limtongia was newly suggested for Zygozyma smithiae, and Limtongia smithiae was proposed as new combination.

In this paper, we described the pharmacological and toxicological studies of three pyrazolone derivatives namely PYZ1: 4-[4-N dimethylamino benzylidine]-3-methyl pyrazolin-5(4H)-one, PYZ2: 4-[2-chlorobenzylidine]-3-methylpyrazolin-5(4H)-one and PYZ3: 4-[benzylidine]-3-methylpyrazolin-5(4H)-one derivatives. Analgesic, anti-inflammatory and antipyretic studies of 3-methyl pyrazolone derivatives at 400 mg/kg, p.o. have shown significant activity as compared to control. Amongst three pyrazolone derivatives, PYZ2 was found to be more active. Based on the result of pharmacological studies, PYZ2 was selected for toxicological studies. Acute toxicity studies revealed that methyl pyrazolone derivatives are non-toxic in rats up to 5000 mg/kg, p.o. The subacute toxicity study of PYZ2 showed that decrease in Hb content, RBC and WBC count. In biochemical analysis level of blood glucose and bilirubin reduced where as AST, ALT and alkaline phosphatase level elevated. Histopathological studies revealed that there was mild toxicity on liver and kidney at 1000 mg/kg, p.o.

Recent changes in the epidemiology of candidiasis highlighted an increase in non-Candida albicans species emphasizing the need for reliable identification methods. Molecular diagnostics in fungal infections may improve species characterization, particularly in cases of the closely related species in the Candida complexes. We developed two PCR/restriction fragment length polymorphism assays, targeting either a part of the intergenic spacer 2 or the entire intergenic spacer (IGS) of ribosomal DNA using a panel of 270 isolates. A part of the intergenic spacer was used for discrimination between C. albicans and C. dubliniensis and between species of the C. glabrata complex (C. glabrata/C. bracarensis/C. nivariensis). The whole IGS was applied to C. parapsilosis, C. metapsilosis, and C. orthopsilosis, and to separate C. famata (Debaryomyces hansenii) from C. guilliermondii (Pichia guilliermondii) and from the other species within this complex (ie, C. carpophila, C. fermentati and C. xestobii). Sharing similar biochemical patterns, Pichia norvegensis and C. inconspicua exhibited specific IGS profiles. Our study confirmed that isolates of C. guilliermondii were frequently misidentified as C. famata. As much as 67% of the clinical isolates phenotypically determined as C. famata were recognized mostly as true P. guilliermondii. Conversely, 44% of the isolates initially identified as C. guilliermondii were corrected by the IGS fingerprints as C. parapsilosis, C. fermentati, or C. zeylanoides. These two PCR/restriction fragment length polymorphism methods may be used as reference tools [either alternatively or adjunctively to the existing ribosomal DNA (26S or ITS) sequence comparisons] for unambiguous determination of the Candida species for which phenotypic characterization remains problematic. (J

The PCR amplification and subsequent restriction analysis of the ribosomal region spanning the internal transcribed spacers ͑ITS1 and ITS2͒ and the 5.8S rRNA gene is applied to the identification of yeasts belonging to the genus Candida. This methodology has previously been used for the identification of some species of this genus, but in the present work this application has been applied to the identification and characterisation of a greater number of species of the genus Candida, with a special survey of species of clinical and biotechnological interest. Among the species of the genus Candida, the high variability observed, both in the length of the amplified region ͑ranging between 390 and 900 bp͒ and in their restriction patterns, allows the unequivocal identification to the species level, with the exception of the group of species that comprises C. membranifaciens, C. conglobata, C. atlantica, C. atmosphaerica, and C. oleophila, that required the sequencing of the D1/D2 domain of the 26S rRNA gene or the 5.8S-ITS region for their proper differentiation. The 5.8S-ITS restriction analysis also failed in the differentiation of species within the pairs C.aaseri ⁄ C.butyri, C.fructus ⁄ C.musae, C.santamariae var. santamariae/C. beechii and C. zeylanoides/C. krissii. In this case, the high sequence similarities obtained for their 26S D1/D2 domain and the 5.8S-ITS region indicate that each pair of species should be considered as a single species. The main purpose of this work is to generate a database for a high number of yeast species, of both biotechnological and clinical interest, and to facilitate their easy, fast, and reliable identification. The present work improves the database available online at the IATA web page ͑http://motor.edinfo.es/iata/͒ with the patterns of 75 species belonging to the genus Candida.

Dialkylamino-3-coumarinyl)pyrazolines and their julolidine analog have been synthesized and proved as new effective photogenerators of acidity. Having D-p-linker ÀA conjugated system of pushpull type these compounds possess intensive absorption in visible region of electronic spectra, high photosensitivity and undergo photodehydrogenation with proton elimination in the presence of CCl 4 or C 2 Cl 6 with a high speed.

Purpose: To synthesize and characterize novel hydroquinone compounds that exhibit an aspirin-like pharmacological profile devoid of ulcerogenic side effects. Methods: Two novel hydroquinone derivatives, viz, 2,5-bis(piperidinomethyl)hydroquinone and 2,5bis(pyrrolidinomethyl)hydroquinone, were synthesized by refluxing hydroquinone, paraformaldehyde and secondary amines (piperidine or pyrrolidine) in ethanol. The structures were authenticated by infrared (IR) spectroscopy, elemental analysis, mass spectrometry (MS) and 1H and 13C nuclear magnetic resonance (NMR) spectroscopic techniques. The synthesized derivatives were evaluated for antinociceptive, anti-inflammatory and antipyretic activities along with gastric-ulcerogenicity using wellknown testing paradigms. Aspirin served as reference standard. Results: The newly synthesized hydroquinone derivatives, significantly attenuated tonic visceral chemically-induced nociception at 10 mg/kg (p < 0.01, p < 0.001), 20 and 40 mg/kg (p < 0.001), inhibited the temporal-inflammatory reaction at 50 mg/kg (2-5 h, p < 0.05, p < 0.001), 100 and 150 mg/kg (1-5 h, p < 0.05, p < 0.01, p < 0.001) in addition to alleviating the febrile-response at test doses during 0.5 h (p < 0.05, p < 0.01, p < 0.001), 1 and 1.5 h (p < 0.001) of the study period. The synthesized compounds exhibited improved gastric tolerability profile since they were devoid of aspirin-associated biochemical and ulcerative changes. The in silico studies predicted high binding affinity of the hydroquinone derivatives to the active site of the cyclooxygenase 2 (COX-2) enzyme. Conclusion: The synthesized hydroquinone compounds possess analgesic, antipyretic and antiinflammatory properties with low gastric-ulcerogenic potential. This may be credited to preferential inhibition of the COX-2 enzyme and the beneficial basic rather than acidic chemical nature of the compounds. However, further molecular studies are required to substantiate these findings.

Scolytine bark beetles are the most destructive pests of conifers; they sometimes aggregate in such large numbers that they actually kill their hosts. They maintain close relationships with yeasts and fungi, in particular those that are assumed to aid in digestive, detoxification processes and pheromone production. In this study, 403 yeast strains were isolated from the guts, ovaries, eggs and frass of nine bark beetle species in the genus Dendroctonus Erichson. The beetles were collected from 10 conifer species at 34 locations in Mexico, Guatemala and the USA. Yeast identification was based on partial DNA sequences from 18S rDNA, 26S rDNA and internal transcribed spacer (ITS1), as well as morphological and physiological characteristics. A combined phylogenetic analysis delimited 11 clades with sequences similar to Candida arabinofermentans, C. ernobii, C. membranifaciens (including C. lessepsii, Pichia mexicana and P. scolyti), C. oregonensis, C. piceae, Kuraishia capsulata (including K. capsulata and K. cf. molischiana), Pichia americana, P. canadensis, P. glucozyma, P. guilliermondii and an undescribed species of Candida. Nucleotide divergences between the major clades were at least 5% while, with the exception of 30 isolates, yeasts within clades differed from named reference species at fewer than 1% of the nucleotide sites. There do not appear to be obligate relationships between particular yeasts and specific anatomical partitions, nor between particular yeasts and bark beetle species. Some yeasts do appear to be preferentially associated with bark beetles feeding on different conifer genera and therefore host plant defences may limit yeast community diversity in Dendroctonus.

Background and Aim: Milk is a food of high nutritional value, which occupies an undeniable place in the human food ration, but is an ideal medium for microbial growth. This study aims to assess the hygienic quality of local raw and fermented milk from the Liptako-Gourma region in Niger. Materials and Methods: We performed physical and bacteriological analyses on 330 samples of bovine milk from local breeds, including 110 individual milk samples (per cow), 110 fermented milk samples, and 110 blended milk samples. The microbiological parameters were determined using standard methods. Results: The physical analysis revealed temperatures during sample collection for all milk types between 35.2°C and 37.8°C. The average pH of fermented milk varied between 3.16 and 4.92 and those of individual and blended raw milks between 5.42 and 6.98. The titratable acidity varied from 15° to 18.1°D for raw milk and between 59° and 122°D for fermented milk. The average density of individual and blended...

Recent changes in the epidemiology of candidiasis highlighted an increase in non-Candida albicans species emphasizing the need for reliable identification methods. Molecular diagnostics in fungal infections may improve species characterization, particularly in cases of the closely related species in the Candida complexes. We developed two PCR/restriction fragment length polymorphism assays, targeting either a part of the intergenic spacer 2 or the entire intergenic spacer (IGS) of ribosomal DNA using a panel of 270 isolates. A part of the intergenic spacer was used for discrimination between C. albicans and C. dubliniensis and between species of the C. glabrata complex (C. glabrata/C. bracarensis/C. nivariensis). The whole IGS was applied to C. parapsilosis, C. metapsilosis, and C. orthopsilosis, and to separate C. famata (Debaryomyces hansenii) from C. guilliermondii (Pichia guilliermondii) and from the other species within this complex (ie, C. carpophila, C. fermentati and C. xestobii). Sharing similar biochemical patterns, Pichia norvegensis and C. inconspicua exhibited specific IGS profiles. Our study confirmed that isolates of C. guilliermondii were frequently misidentified as C. famata. As much as 67% of the clinical isolates phenotypically determined as C. famata were recognized mostly as true P. guilliermondii. Conversely, 44% of the isolates initially identified as C. guilliermondii were corrected by the IGS fingerprints as C. parapsilosis, C. fermentati, or C. zeylanoides. These two PCR/restriction fragment length polymorphism methods may be used as reference tools [either alternatively or adjunctively to the existing ribosomal DNA (26S or ITS) sequence comparisons] for unambiguous determination of the Candida species for which phenotypic characterization remains problematic. (J

Four halotolerant yeast strains, M21T, M34-1, HS054 and D41, were isolated from various foods in South-East Asia. These isolates were most closely related to Pichia anomala, with which each strain had from zero to two differences in the 26S rDNA D1/D2 domain nucleotide sequence; for this reason, they were thought to be the same as, or sister species of, P. anomala. Of the four yeast isolates, only one strain, M21T, had an 18S rDNA sequence that differed from those of P. anomala IFO 10213T and the other three isolates, having 20 substitutions and two gaps. Strain M21T showed lower cation (Li+) tolerance (⩽0·3 M LiCl) than P. anomala IFO 10213T or the other three strains (⩽0·5 M LiCl). Furthermore, the DNA–DNA hybridization data indicated that M21T was clearly distinct from P. anomala IFO 10213T and the other three isolates. The ability of strain M21T to assimilate d-arabinose distinguished it from P. anomala IFO 10213T and the other three isolates; it also differed in that it was abl...

The quality evaluation of white soft cheese in the Egyptian markets is an urgent need to create awareness among population about the existing situation and to protect the consumer&#39;s health and rights. Thus, the aim of this work was to determine AFM levels and evaluate microbiological properties in 1 commercially available white soft cheese collected from local markets as well as to evaluate the manufactured control samples using the Hazard Analysis Critical Control Points (HACCP) system. Results revealed that AFM1 concentration was detected in almost half of the samples (46.66%) with an overall mean of 14.39 ng/kg and was detected in 24 samples at levels below 1.0 ng/kg and in 15 samples at levels ranging from 10 to 50 ng/kg, whereas about 3 samples were higher than 50 ng/kg. All positive samples of white cheeses exceeded the Egyptian regulations (free from AFM ). The microbiological analysis revealed that all samples contained high 1

The present study was designed to investigate the gastroprotective, analgesic, and antipyretic effects of curcumin (Cur), the major constituent of turmeric. Acetylsalicylic acid (ASA) was used in this study as a standard drug for comparison. The analgesic activity was measured using the Hot-Plate Test. The antipyretic and antiulcer effects were assessed using yeast-induced pyrexia and gastric ulceration, respectively. Curcumin (100 mg/kg) injected intra-peritoneally 1 hr prior to the Hot-Plate Test showed significant analgesic activity expressed by both parameters: an increase in latency time and a reduction in paw licking as compared to the controls. In the animal model of pyrexia, curcumin (100 mg/kg injected intra-peritoneally) exhibited a significant reduction in the rectal temperature after 1 hr, 2 hrs, 4 hrs, and 5 hrs of treatment, indicating the antipyretic effect of curcumin. Rats with orally administered curcumin (200 mg/kg) did not show any lesions on the inner lining of ...

Cheese is a dairy product with a high nutritional value, a unique flavor, aroma, a long shelf life, and a very different variety. Cheese may be contaminated with harmful microorganisms which may pose a risk to public health during the raw material procurement and production stages. This study investigated the microbiological quality of 60 white cheese samples, marketed in Afyonkarahisar province. The mean levels of total mesophilic aerobic bacteria, Enterobacteriaceae, coliform, Esherichia coli, coagulase-positive staphylococci, and yeast-mold were determined as 2,1x10 5 , 1,4x10 4 , 2,1x10 3 , 4,5x10 2 , 4,1x10 2 and 3,8x10 3 CFU/g, respectively. Also, Escherichia coli was found in 10% (6/60) of the samples between the level of 10 2-10 5. Consequently, the microbiological quality was found to be insufficient in some of the samples examined. Therefore, it is recommended to take necessary hygienic measures in raw material supply. In addition, necessary precautions should be taken in all stages of cheese production and attention should be paid to the personnel and equipment hygiene. Contribution/Originality: This study investigated the microbiological quality of 60 white cheese samples, marketed in Afyonkarahisar province. 1. INTRODUCTION Animal-origin food products play an important role regarding the balanced and adequate nutrition of people (Kırdar, 2001). Milk and milk products are among the important foods of animal origin. Cheeses are important in nutrition since their structure comprises proteins, fats, vitamins and minerals (Fox & Mcsweeney, 2004). Approximately, 4000 types of cheese are available in the world, over 110 types of which can be found in our country (Kamber & Terzi, 2007). White cheese assumes the first place between the produced and consumed cheese assortment in our country. According to Turkish Food Codex, white cheese 'with proper processing technique and using rennet enzyme coagulation of milk and the curd which is made from coagulated milk, could be produced in fresh and mature form in the production stage, which shows variety-specific characteristics (Turkish Food Codex, 2015). On the other hand, milk and dairy products are also important sources of foodborne infections and intoxications, in addition to their positive properties, cheese obtained from raw milk may pose a great risk to public health (Mead et al., 1999). Bacterial contamination of raw milk can be caused by different sources such as air, dairy equipment, animal feeds, soil, and feces (Coorevits et al., 2008). In addition, the use of raw milk with low

The identification of twenty four yeast isolates from cheese performed with API ID 32C simplified assimilation test and restriction fragments length polymorphism (PCR-RFLP) of ITS1-5.8S rDNA-ITS2 region was compared. The accordance of the results of both methods for eleven strains identified as Candida famata, five strains as C. lipolytica and four strains as C. sphaerica was observed. However, the other four yeast isolates classified with API as C. sphaerica revealed amplification products as well as restriction patterns characteristic for C. famata species.

The main objective of this research was to study the safety aspects of two Sudanese street foods, namely: Tamia (Falafil) and Dakwa (peanut butter) collected from different sources in Khartoum State. Samples for Tamia were obtained from Albait Alssory-Khartoum North, College of Agricultural Studies-Sudan University of Science and Technology and Omdurman market. Samples for Dakwa were obtained from Shambat Central Market and crusher machine. Microbial and chemical characteristics were evaluated. The total bacterial count (TBC) (cfu/g) in Tamia was 5.6x10 , 3.9x10 and 4.9x10 in samples of Albait Alssory, College and Omdurman 3 4 5 market, respectively. Coliforms and moulds and yeasts in Tamia were not detected in Albait Alssory and College, while Omdurman market samples recorded 1.7x10 for coliforms and 5.8x 10 for moulds and yeast. 2 Pathogenic bacteria (E. coli, Staph. aureus and Salmonella) were not detected in all samples except Staph. aureus were detected in Omdurman market (3.9x10). TBC in Dakwa were 5.4x10 , 5.6x10 and 6.4x10 in crusher 2 3 4 4 machine, Shambat market and Omdurman market, respectively. Coliforms were detected in all sources of Dakwa, while moulds and yeasts were detected only in Omdurman market. Albait Alssory was the only source of Dakwa free of pathogenic bacteria. Lead (Pb) in Tamia was 3.04 (ppb) in Albait Alssory and was not detected in the other two sources (College or Omdurman market). Alaminium (Al) and peroxide values were found in all sources of Tamia. Pb in Dakwa was 3.20 in Shambat market and was not detected in crusher machine and Omdurman market. Al in Dakwa was present in all sources while peroxide value was absent in the crusher machine.

Scolytine bark beetles are the most destructive pests of conifers; they sometimes aggregate in such large numbers that they actually kill their hosts. They maintain close relationships with yeasts and fungi, in particular those that are assumed to aid in digestive, detoxification processes and pheromone production. In this study, 403 yeast strains were isolated from the guts, ovaries, eggs and frass of nine bark beetle species in the genus Dendroctonus Erichson. The beetles were collected from 10 conifer species at 34 locations in Mexico, Guatemala and the USA. Yeast identification was based on partial DNA sequences from 18S rDNA, 26S rDNA and internal transcribed spacer (ITS1), as well as morphological and physiological characteristics. A combined phylogenetic analysis delimited 11 clades with sequences similar to Candida arabinofermentans, C. ernobii, C. membranifaciens (including C. lessepsii, Pichia mexicana and P. scolyti), C. oregonensis, C. piceae, Kuraishia capsulata (including K. capsulata and K. cf. molischiana), Pichia americana, P. canadensis, P. glucozyma, P. guilliermondii and an undescribed species of Candida. Nucleotide divergences between the major clades were at least 5% while, with the exception of 30 isolates, yeasts within clades differed from named reference species at fewer than 1% of the nucleotide sites. There do not appear to be obligate relationships between particular yeasts and specific anatomical partitions, nor between particular yeasts and bark beetle species. Some yeasts do appear to be preferentially associated with bark beetles feeding on different conifer genera and therefore host plant defences may limit yeast community diversity in Dendroctonus.

Healthy food production is the main priority of food technology and dairy products which plays an important role in the household food basket. Traditional dairy products take a large part in the market and sometimes are causing transmission of various diseases to humans, particularly in the cheese market. Siahmazgi cheese is a type of traditional cheese, produced from raw milk in the north of Iran, and is getting increasingly popular. In this research, physicochemical, microbial and sensory characteristics of five samples of bought traditional cheese and five samples of manufactured pasteurized cheese are studied on days 0, 30, and 60. The result of the physicochemical survey shows the solids content, pH, and fat contents of traditional and pasteurized cheese samples decreased over 60 days, but the difference was not always statistically significant. On the other hand, the protein and salt content of traditional and pasteurized cheeses significantly increased over 60 days (p<0.05). The protein and salt content were higher in traditional samples. The results also revealed that pasteurization eliminates pathogens from cheese whereas most of the traditional Siahmazgi cheeses were infected by Coliform, Escherichia coli, yeast-mold, and staphylococcus. There was not observed salmonella infection in any samples of traditional Siahmazgi cheeses. Sensory evaluation between pasteurized and traditional Siahmazgi cheese shows that only texture has significantly changed over 60 days and the other sensory features were the same.

Two yeasts isolated from laboratory reared adult coffee berry borers [ Hypothenemus hampei (Ferrari) (Coleoptera: Scolytidae)] and from insects collected in the field in Colombia were identified as Pichia burtonii Boidin and Candida fermentati (Saito) Bai, based on sequencing of the nuclear large subunit 26S rDNA variable D1/D2 domain. Liquid culture experiments using P. burtonii in media containing different caffeine levels indicated that caffeine levels in a range found within coffee seeds can retard yeast growth. HPLC analysis shows that P. burtonii does not break down caffeine.

Thrust in the preparation of new heterocyclic molecules is increasing due to their proven significant biological activities. The paramount importance of heterocycles such as indoles and their derivatives in natural product chemistry and pharmacology constantly drives the search for the new procedures for their construction and also for the preparation of variety of their derivatives to exploit their useful biological activities. A tetrahydrocarbazole had been synthesized by reacting cyclohexanones with substituted hydrazine hydrochlorides in the presence of glacial acetic acid to produce N-(1,2,3,4-tetrahydrocarbazole) which later used to synthesize compounds with different heterocyclic new rings containing, sulfur and nitrogen in structures i.e., substituted-N-(1,2,3,4-tetrahydrocarbazole) which were characterized by IR, 1 H and 13 C NMR spectral studies and these structures were investigated for analgesic and anti inflammatory activity.

soft cheese pseudomonas antibiotic sensitivity potassium sorbate One hundred and twenty five random samples of white soft cheese, fifty of each Damietta, kariesh and twenty five of Feta cheese were collected from street vendors, supermarkets and groceries at Damietta province. All samples were examined chemically and sensory and proved to be fit for human consumption. The prevalence of pseudomonas spp. was 68% and 58% with mean count 9.02x10 4 ±2.87x10 4 and 2.43x10 5 ±9.32x10 4 for Damietta and kariesh cheese respectively. Pseudomonas spp were not detected in Feta cheese. The most prevalent spp was p. fluorescens (35.14% and 45.5%) while p.aeruginosa (21.26% and 15.2%); p. putida (27% and 18.2%) and p. stutzeri (16.21%and 21.21%) for Damietta and kariesh cheese respectively. All strains of p.aeruginosa showed 100% resistance to Ampicillin, Ampicillin/sulbectam, Cefazolin, Tigecyclin, Nitrofurantoin and Trimethoprim / sulfamethoxazole followed by Cefatriaxone and Cefepim 23% while Imipenem, Meropenem , Amikacin , Gentamycin and Tobramycin showed 100% sensitivity followed by Ciprofloxacin and Moxifloxacin 69.23%. Viability of p.aeruginosa during manufacture and storage of Damietta cheese was studied, using 0.1% Potassium sorbate with 10% Sodium chloride and cold storage of cheese showed great inhibitory effect on p.aeruginosa at 14 th day. The public health significance of these finding were discussed.

Purpose: To synthesize and characterize novel hydroquinone compounds that exhibit an aspirin-like pharmacological profile devoid of ulcerogenic side effects. Methods: Two novel hydroquinone derivatives, viz, 2,5-bis(piperidinomethyl)hydroquinone and 2,5bis(pyrrolidinomethyl)hydroquinone, were synthesized by refluxing hydroquinone, paraformaldehyde and secondary amines (piperidine or pyrrolidine) in ethanol. The structures were authenticated by infrared (IR) spectroscopy, elemental analysis, mass spectrometry (MS) and 1H and 13C nuclear magnetic resonance (NMR) spectroscopic techniques. The synthesized derivatives were evaluated for antinociceptive, anti-inflammatory and antipyretic activities along with gastric-ulcerogenicity using wellknown testing paradigms. Aspirin served as reference standard. Results: The newly synthesized hydroquinone derivatives, significantly attenuated tonic visceral chemically-induced nociception at 10 mg/kg (p < 0.01, p < 0.001), 20 and 40 mg/kg (p < 0.001), inhibited the temporal-inflammatory reaction at 50 mg/kg (2-5 h, p < 0.05, p < 0.001), 100 and 150 mg/kg (1-5 h, p < 0.05, p < 0.01, p < 0.001) in addition to alleviating the febrile-response at test doses during 0.5 h (p < 0.05, p < 0.01, p < 0.001), 1 and 1.5 h (p < 0.001) of the study period. The synthesized compounds exhibited improved gastric tolerability profile since they were devoid of aspirin-associated biochemical and ulcerative changes. The in silico studies predicted high binding affinity of the hydroquinone derivatives to the active site of the cyclooxygenase 2 (COX-2) enzyme. Conclusion: The synthesized hydroquinone compounds possess analgesic, antipyretic and antiinflammatory properties with low gastric-ulcerogenic potential. This may be credited to preferential inhibition of the COX-2 enzyme and the beneficial basic rather than acidic chemical nature of the compounds. However, further molecular studies are required to substantiate these findings.

In order to develop an efficient an system for waste water pretreatment, the isolation of indigenous population of microorganisms from pharmaceutical waste water was done. We obtained pure cultures of 16 yeast isolates that differed slightly in colony morphology. Ten out of 16 isolates efficiently reduced COD in pharmaceutical waste water. Initial physiological characterization failed to match the 10 yeast isolates to either Pichia anomala or Pichia ciferrii. Restriction analysis of rDNA (rDNA-RFLP) using three different restriction enzymes: HaeIII, MspI and CfoI, showed identical patterns of the isolates and Pichia anomala type strain. Separation of chromosomal DNAs of yeast isolates by the pulsed field gel electrophoresis revealed that the 10 isolates could be grouped into 6 karyotypes. Growth characteristics of the 6 isolates with distinct karyotypes were then studied in batch cultivation in pharmaceutical waste water for 80 hours.

The identification of twenty four yeast isolates from cheese performed with API ID 32C simplified assimilation test and restriction fragments length polymorphism (PCR-RFLP) of ITS1-5.8S rDNA-ITS2 region was compared. The accordance of the results of both methods for eleven strains identified as Candida famata, five strains as C. lipolytica and four strains as C. sphaerica was observed. However, the other four yeast isolates classified with API as C. sphaerica revealed amplification products as well as restriction patterns characteristic for C. famata species.

The genetic diversity of the types or authentic strains of 20 facultative synonyms of Pichia membranifaciens (E.C. Hansen) E.C. Hansen was revealed on the basis of large-subunit (26S) rDNA D1/D2 domain and internal transcribed spacer region sequencing and electrophoretic karyotyping. At least five separate species were recognized among the strains studied. Fourteen strains were confirmed to belong to P. membranifaciens. Strain CBS 241, an authentic strain of Zygosaccharomyces chevalieri Guilliermond var. fermentati Saito, should be assigned to Pichia manshurica Santa María. Strain CBS 243, an authentic strain of Zygopichia chiantigiana Castelli, is conspecific with CBS 2287, the type strain of Pichia fluxuum (Phaff & Knapp) Kreger-van Rij. Strain CBS 1367, the type of Zygosaccharomyces bisporus Anderson, belongs to Pichia kluyveri Bedford var. kluyveri. Strain CBS 989, the type of Cryptococcus californicus Anderson & Skinner, represents a distinct species, for which a new combination, Candida californica comb. nov., is proposed. The taxonomic status of strains CBS 189, the type of Pichia calliphorae Klöcker, and CBS 214, the type of Pichia derossii Castelli, remain to be studied further. Their D1/D2 sequences and chromosomal DNA banding patterns were similar to those of P. membranifaciens, but their internal transcribed spacer sequences differed significantly.

Background and Aim: Milk is a food of high nutritional value, which occupies an undeniable place in the human food ration, but is an ideal medium for microbial growth. This study aims to assess the hygienic quality of local raw and fermented milk from the Liptako-Gourma region in Niger. Materials and Methods: We performed physical and bacteriological analyses on 330 samples of bovine milk from local breeds, including 110 individual milk samples (per cow), 110 fermented milk samples, and 110 blended milk samples. The microbiological parameters were determined using standard methods. Results: The physical analysis revealed temperatures during sample collection for all milk types between 35.2°C and 37.8°C. The average pH of fermented milk varied between 3.16 and 4.92 and those of individual and blended raw milks between 5.42 and 6.98. The titratable acidity varied from 15° to 18.1°D for raw milk and between 59° and 122°D for fermented milk. The average density of individual and blended...