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A photothrombotic model of small early ischemic infarcts in the rat brain with histologic and MRI correlation.

Over the last two decades several studies have suggested the role of photothrombotic occlusion of cerebral microvessels using rose bengal, resulting in small strokes in rodents that resemble those in humans. This paper describes such a photothrombotic method of acute small stroke induction in rats with histopathologic and in vivo magnetic resonance imaging (MRI) observations from 3 to 6 h after irradiation, which is homologous to a human autopsy specimen. Utilizing 30 min of irradiation with minimal beam intensity (0.1 W/cm(2)) cold white light in conjunction with 20 mg of intravenous (iv) rose bengal as a rapid infusion, small infarcts were induced photochemically in the frontal lobes of six rats. The infarcts showed a consistent pattern on histologic and in vivo MR sections when examined within 7 h or less of irradiation. Both MRI and histologic sections were comprised of (a) a superior zone of infarcted neurons, (b) a middle curvilinear transition zone of edema on MRI and histologically vacuolated neuropil, and (c) an inferior zone of normal neurons. Shorter duration water-sensitive (T2)- and postgadolinium longer duration (T1)-weighted signal decay images both showed a curvilinear hyperintense transition zone of edema. The mean infarct and transition zone areas measured from the histologic sections were comparable to those measured on the MRI. The infarct model described above allows in vivo observations using MRI with the potential for use in testing putative neuroprotective agents. As demonstrated by a comparison with the histologic features of such infarcts in surgical and autopsy brain specimens, the model is relevant to acute human ischemic infarcts.

Animals↗

Structural and functional degradation of Ca2+:Mg2+-ATPase rich sarcoplasmic reticulum vesicles photosensitized by erythrosin B.

Erythrosin B (Red Dye No. 3) and Rose Bengal photosensitize the destruction of the Ca2+:Mg2+-ATPase pump protein in sarcoplasmic reticulum (SR) vesicles with respective quantum efficiencies of (1.53 +/- 0.19) X 10(-3) and (1.25 +/- 0.18) X 10(-3). Damage to vesicle function was assayed by measurements of increases in passive Ca2+ permeability. Rates of passive Ca2+ movement into the SR lumen were increased by dye photosensitization in proportion to radiation absorbed. Active Ca2+ transport into SR vesicles was blocked independent of radiation absorbed by Erythrosin B and Rose Bengal at free concentrations of 0.69 microM and 1.16 microM, respectively. The photochemical lability of the Ca2+ pump protein and alterations in passive and active Ca2+ transport may be dependent on the concentration of the dye in the membrane. The photosensitization results may have implications with respect to the suitability of Erythrosin B usage in vivo, since the brightness of our irradiation source is comparable to that of sunlight at 480 nm.

Animals↗

Diagnostic tests in patients with symptoms of keratoconjunctivitis sicca.

We compared the relative value of the tear film breakup time, Schirmer test, lactoferrin immunologic assay and rose bengal staining in 156 patients with varying severity of symptoms characteristic of keratoconjunctivitis sicca and in 39 controls. Statistical analysis showed that in patients with minimal ocular irritation the Schirmer test in combination with the lactoferrin immunologic assay provided an optimal balance between high test sensitivity and low false-positive rates. In patients with moderate to severe ocular burning, foreign body sensation, or pain but with no systemic symptoms of dry mouth or arthritis, and in patients with both ocular and systemic complaints, the lactoferrin assay alone provided statistically significant results. Tear film breakup time and staining with rose bengal were not useful and the Schirmer test had limited value in measuring the rate of tear secretion in these patients.

Adult↗

Evidence for essential histidine residues in bovine-liver mitochondrial monoamine oxidase.

Ethoxyformic acid anhydride, amino-1H-tetrazole, and photooxidation in the presence of rose bengal, which are reagents known to react with histidine residues of proteins, were shown to inactivate monoamine oxidase. Ethoxyformic acid anhydride reacted with about 6 histidine residues per 100 000 g of protein under the experimental conditions adopted and completely inactivated the enzyme. However, NH2OH reactivated the ethaxyformic acid derivative t only. Since NH2OH specifically deacylates N-ethoxyformylimidazole, it was shown that at least some of the histidine residues are essential for activity. In addition, photooxidation experiments in the presence of 0.01% rose bengal confirmed that only histidine residues of bovine hepatic monoamine oxidase are destroyed under the designated experimental conditions. About 9 histidine residues per 100 000 g of protein were destroyed during the photooxidation experiments. In the presence of substrate, kynuramine or benzylamine, only 7 histidine residues were destroyed, which indicates that 2 histidine residues per 100 000 g of protein are essential for activity.

Amino Acids↗

Lid wiper epitheliopathy and dry eye symptoms.

OBJECTIVES: The lid wiper is defined as that portion of the marginal conjunctiva of the upper eyelid that wipes the ocular surface during blinking. The purpose of this study was to investigate whether lid wiper epitheliopathy occurred with patients who reported dry eye symptoms, yet had normal fluorescein breakup time (FBUT) and Schirmer test values and an absence of fluorescein corneal staining. METHODS: One hundred patients were divided into two groups based on the presence or absence of dry eye symptoms, as determined with the Standard Patient Evaluation of Eye Dryness questionnaire. Other criteria for admission to both groups were FBUT of 10 seconds or more, Schirmer test value of 10 mm or more, and absence of fluorescein corneal staining. After instillation of fluorescein and rose bengal dyes, the lid wipers of 50 asymptomatic and 50 symptomatic patients were graded for staining from grade 0 (absent) to grade 3 (severe). RESULTS: Of the symptomatic patients, 76% had staining of the lid wiper: 44%, grade 1; 22%, grade 2; and 10%, grade 3. Of the asymptomatic patients, 12% had staining; 8%, grade 1; 4%, grade 2; and 0%, grade 3. The difference in prevalence of lid wiper staining between the symptomatic and asymptomatic groups was significant (P<0.0001). CONCLUSIONS: Lid wiper epitheliopathy, diagnosed by staining with fluorescein and rose bengal dyes, is a frequent finding when symptoms of dry eye are experienced in the absence of routine clinical dry eye findings.

Adult↗

Mucus fishing syndrome: case report and new treatment option.

BACKGROUND: Mucus fishing syndrome (MFS) is a cascading cyclic condition characterized by continuous extraction of mucous strands from the eye. It is usually initiated by ocular irritation. In response to irritation, ocular surface cells produce excess mucus. A "snow balling" cycle begins when the patient extracts ("fishes") excess mucus from the ocular surface, thereby causing further irritation and a more-profound mucous discharge. To date, treatment includes eliminating the initiating element and educating the patient not to touch the eye when extracting the excess mucus, CASE REPORT: Presented is a case of mucus fishing syndrome initiated by dry eye. The patient's diagnosis, MFS, was identified by persistent mucous discharge, his admittance and demonstration of digitally extracting mucus from the ocular surface, and a characteristic rose bengal staining pattern. The conventional treatment initiated by using artificial tears for the dry eye condition and educating the patient not to touch the ocular surface did not provide relief from the excess mucous discharge. Therefore, a new approach to treatment was pursued. In order to break the cycle, a mucolytic agent and an antihistamine-mast cell stabilizer were prescribed, until the ocular surface healed. After treatment, the patient reported alleviation of symptoms and demonstrated improvement in ocular surface integrity by a profound reduction in rose bengal staining. CONCLUSION: Mucus fishing syndrome is challenging to resolve with conventional treatment because it requires a certain level of psychological tolerance and perseverance from the patient. By eliminating the present mucus and diminishing mucous production pharmacologically, the practitioner is able to remove the stimulus for digital extraction and thus accelerate ocular surface healing. We present a proposed new treatment option for patients who are refractory to conventional treatments.

Cystine↗

Correlation between impression cytology and tear function parameters in Sjögren's syndrome.

We have compared clinical (Schirmer I test, BUT and rose bengal staining), laboratory (lysozyme and lactoferrin tear levels) and histological tests (impression cytology) in 165 eyes from 85 patients with primary Sjögren's syndrome and in 80 eyes from 40 control subjects. Impression cytology can provide the location of cellular alterations on the ocular surface and information on the severity of the disease. The upper bulbar and interpalpebral areas from patients with primary Sjögren's syndrome were shown to have cellular alterations early in the course of the disease, while the lower bulbar and lower palpebral areas were only affected in severe cases. Statistical analysis has shown that impression cytology and rose bengal staining are the most specific and sensitive methods for the diagnosis of primary Sjögren's syndrome.

Adult↗

Tear function and ocular surface changes in keratoconus.

PURPOSE: To describe the ocular surface disorder in patients with keratoconus. DESIGN: A prospective, case-controlled study. PARTICIPANTS: Seventy-five eyes of 38 patients with keratoconus seen at Uludag University School of Medicine, Department of Ophthalmology, from March 2000 through April 2001, and 80 eyes of 40 normal control subjects were studied. INTERVENTION: The subjects underwent routine ophthalmic examinations, corneal sensitivity measurements, Schirmer test, tear film breakup time (BUT), fluorescein and rose bengal staining of the ocular surface, and conjunctival impression cytology. MAIN OUTCOME MEASURES: Patients and control subjects were compared for corneal sensitivity, tear function, ocular surface staining parameters, goblet cell density, and squamous metaplasia grade. The relation of these parameters to the severity of keratoconus progression was also investigated. RESULTS: The mean corneal sensitivity was significantly lower in keratoconus patients compared with control subjects (P < 0.001). The BUT values were also significantly lower in the keratoconus group. Patients with keratoconus had significantly higher fluorescein and rose bengal staining scores (P < 0.001). Corneal sensitivity and tear function changes seemed to get worse with advanced stages of keratoconus. Impression cytology showed goblet cell loss and conjunctival squamous metaplasia, both of which again related to the extent of progression of keratoconus. CONCLUSIONS: The ocular surface disease in keratoconus is characterized by disorder of tear quality, squamous metaplasia, and goblet cell loss, all of which seem to relate to the extent of keratoconus progression.

Adolescent↗

Effect of gefarnate on the ocular surface in squirrel monkeys.

PURPOSE: To investigate the ability of gefarnate (geranyl farnesylacetate) to stimulate goblet cell function in the primate eye after a mild alkali injury of the tarsal conjunctiva. METHODS: A bilateral injury was created on the conjunctival surface of the lower eye lid of squirrel monkeys by means of a 30-second application of a 4-mm diameter piece of filter paper wetted with 0.5% NaOH. Gefarnate drops (1%) were administered to one eye of each monkey and vehicle alone in the contralateral eye six times a day, 5 days a week for 4 weeks. Slit-lamp biomicroscopy, impression cytology staining of the ocular surface, fluorescein and rose bengal staining, and Western blot for mucin were performed before injury and weekly thereafter. Light microscopy was used to evaluate the lower conjunctiva. RESULTS: Topical application of gefarnate was not associated with any adverse ocular surface effects. Goblet cell repopulation after injury was significantly greater in the gefarnate-treated eyes compared with the vehicle-treated eyes. In the gefarnate-treated eyes, tear mucin content was significantly greater at 1 week after injury. Fluorescein staining was significantly reduced at 3 weeks after injury, and rose bengal staining was significantly reduced in the area of the wound at 2 weeks in the gefarnate-treated eyes compared with the vehicle-treated eyes; at other times, conjunctival staining in the two groups of eyes was not significantly different. CONCLUSIONS: Gefarnate promotes goblet cell repopulation and increases mucin production after a conjunctival injury. No adverse affects of the treatment were found. Thus, this agent may be useful in conditions that diminish goblet cell function.

Animals↗

Clinical characteristics of conjunctivochalasis with or without aqueous tear deficiency.

AIM: To show characteristic ocular surface findings caused by conjunctivochalasis (CCh) in dry eye patients with or without aqueous tear deficiency (ATD). DESIGN: Comparative non-interventional cases. PATIENTS AND METHODS: Clinical data of five ATD patients without CCh (group A), eight CCh patients with ATD (group B), and eight CCh patients without ATD (group C) were retrospectively reviewed. Presence or absence of CCh was determined by fluorescein staining to outline tear meniscus and conjunctival folds with an enhancing filter. Dry eye symptoms, history of subconjunctival haemorrhage, meibum expression, tear break up time, fluorescein and rose bengal staining, and fluorescein clearance test, and other abnormal ocular surface findings were measured. RESULTS: CCh patients were significantly older (p = 0.001). In pure ATD, the principal symptom of dryness became worse as the day progressed. In contrast, blurry vision, burning sensation, and dryness became worse during reading in all CCh patients (p = 0.0008) or worse in the morning upon awakening in the majority patients with CCh only (p = 0.02). Besides the interpalpebral exposure, which was noted in ATD, positive fluorescein or rose bengal staining was noted in the redundant conjunctival folds and the non-exposure zone in CCh (p = 0.0008). Redundant conjunctival folds were present in both lower and upper bulbar conjunctiva, obliterating both lower and upper tear meniscuses, and spatially correlated with anterior migration of the mucocutaneous junction in CCh. Delayed tear clearance was significantly more prevalent in CCh than ATD (p = 0.0008). Vigorous blinking worsened in CCh but not in ATD (p = 0.0008). Lacrimal puncta were swollen in groups B and C, but not in group A (p = 0.04). CONCLUSIONS: CCh is not restricted to the lower bulbar conjunctiva, and contributes to pathogenesis of dry eye by obliterating both lower and upper tear meniscus, causing unstable tear film and by creating delayed tear clearance. Dry eye symptoms were worsened by downgaze during reading and by vigorous blinking. Other characteristic signs including subconjunctival haemorrhage, swollen puncta, anterior migration of the mucocutaneous junction, and patterns of dye staining also help distinguish dry eye associated with CCh from that caused by ATD alone.

Adult↗

Effect of carmellose-based artificial tears on the ocular surface in eyes after laser in situ keratomileusis.

BACKGROUND: Strategies to minimize intraoperative ocular surface disturbance and accelerate postoperative ocular surface recovery are an important consideration to the surgeon. METHODS: Eighteen eyes of 10 patients were randomized to receive intraoperative and postoperative non-preserved balanced salt solution or non-preserved carboxymethylcellulose-based (carmellose) artificial tears. Presence and severity of McMonnies dry eye symptom survey score primary dry eye symptoms, fluorescein break-up time, and rose bengal staining were graded preoperatively and at 1 day, 1 week and 1 month postoperatively. Conjunctival impression cytology of the perilimbal conjunctiva was performed 1 month after surgery to determine goblet cell density. RESULTS: McMonnies dry eye symptoms were reported more frequently for the balanced salt solution group at 1 day, 1 week, and 1 month after surgery. Fluorescein break-up time was significantly greater in the carmellose-based group at 1 day (P < .01), 1 week (P = .04), and 1 month (P < .01) after surgery. The rose bengal staining score was significantly greater at 1 day (P < .01) and 1 week (P < .01) postoperatively for the balanced salt solution group. Mean goblet cell density for the carmellose-based group at 1 month postoperatively was significantly greater than that for the balanced salt solution group (P < .01). CONCLUSIONS: Postoperative and intraoperative use of carmellose-based artificial tears resulted in significantly more rapid recovery of the ocular surface after LASIK when compared to balanced salt solution.

Astigmatism↗

Photodynamic alteration of cornea endothelium. Relation to bicarbonate fluxes and oxygen concentration.

Corneas were mounted in flux chambers and endothelial bicarbonate fluxes were determined following sensitization of endothelial cells with 5 . 10(-6) M rose bengal and exposure to light. Corneas exposed to light demonstrated an increased passive bicarbonate flux compared to corneas not photosensitized. Active bicarbonate flux was reduced after 5 min of light exposure, but not after 1 min of light exposure. The increase in passive bicarbonate flux was prevented by the addition of 200 microgram/ml catalase to the bathing solution; however, catalase had no effect on the photodynamic alteration of active flux. Neither 10 mM ascorbic acid nor 1.012 gram/l glutathione prevented the photodynamically induced increase in passive flux. Perfusion of corneas with 5 . 10(-6) M rose bengal dissolved in a sucrose-substituted Krebs-Ringer bicarbonate solution with a PO2 of 124 +/- 4.0 mmHg and exposed to light swelled at rates more rapid than corneas treated in a similar fashion but perfused with a solution with a PO2 of 20 +/- 4.6 mmHg. This study demonstrated that photodynamically induced corneal endothelial cell alteration results in increased passive bicarbonate flux, a time-dependent decrease in active bicarbonate flux, is oxygen dependent, and is at least in part secondary to H2O2 produced by the dismutation reaction of the superoxide free radical.

Animals↗

Effect of lipo-pro-prostaglandin E1, AS-013 on rat inner ear microcirculatory thrombosis.

We investigated effects of lipo-pro-prostaglandin E1 (lipo-[11alpha, 13E, 15S]-11,15-dihydroxy-9-[1-oxobutoxy]-prosta-8, 13-dien-1-oic acid butyl ester), AS-013 in two models of hearing disturbance and equilibrium dysfunction induced by rat inner ear microcirculatory thrombosis. Inner ear microcirculatory thrombosis was induced by photochemical reaction between systemic injection of Rose Bengal and irradiation of green light to the cochlea and vestibule. Photochemical reaction causes endothelial injury followed by platelet adhesion, aggregation and formation of a platelet- and fibrin-rich thrombus. In the hearing disturbance model, a compound cochlear nerve action potential was recorded by electrocochleography every minute. Photochemical reaction induced inner ear microcirculatory thrombosis, followed by disappearance of the action potential. AS-013 significantly (P<0.05) prolonged time to disappearance of the action potential compared with control group. In the equilibrium dysfunction model, the irradiation to the vestibule was applied for 10 min after Rose Bengal injection. The behavior of rats in the swimming test and nystagmus were observed 24 h after the completion of irradiation. In the swimming test, two of 12 animals treated with AS-013 showed no rotating about their longitudinal axes, which indicates equilibrium dysfunction and the duration of well-balanced swimming prolonged. AS-013 suppressed the appearance of nystagmus. These results suggest that lipo-pro-prostaglandin E1, AS-013 may prevent hearing disturbance and equilibrium dysfunction due to inner ear microcirculatory disorders.

Action Potentials↗

Local cerebral blood flow autoregulation following "asymptomatic" cerebral venous occlusion in the rat.

OBJECT: Maintenance of cerebral blood flow (CBF) autoregulation in the brain is of major importance for patient outcome in various clinical conditions. The authors assessed local autoregulation after "asymptomatic" cortical vein occlusion. METHODS: In Wistar rats, a single cortical vein was occluded photochemically by using rose bengal and fiberoptic illumination. In rats with bilateral carotid artery occlusion, mean arterial blood pressure (MABP) was lowered in 5-mm Hg increments down to 40 mm Hg by using hypobaric hypotension. Local CBF at each pressure level was assessed by performing laser Doppler (LD) scanning at 25 (5 x 5) locations within bilateral cranial windows. In this manner, the lower limit of autoregulation (LLA) was detected. The LLA was 60 mm Hg in both right and left hemispheres in Group A (five rats), in which the animals received illumination without rose bengal and had no venous occlusion. Of the 11 rats that underwent vein occlusion, three developed severe reductions in local CBF and/or a growing venous thrombus and were distinguished as Group C (symptomatic; three rats); from previous work we know that those animals are bound to experience venous infarction. The remaining rats formed Group B (asymptomatic; eight rats). In this group the LLA remained at 60 mm Hg in the left hemisphere without occlusion, whereas, in the right cortex with the occluded vein, the LLA was found to be 65 mm Hg. Below a carotid stump pressure of 25 mm Hg regional CBF in the affected hemisphere dropped more abruptly to a possibly ischemic range than that in the opposite normal hemisphere. CONCLUSIONS: The results of the present study suggest that cerebral venous circulation disorders are manifested via additional pathways, that is, from a partially impaired local autoregulation in the vicinity of the occluded vein, even under conditions in which the vein occlusion itself does not cause brain damage. Care should be taken in the control of blood pressure in patients with this pathological condition.

Analysis of Variance↗

Lid-wiper epitheliopathy and dry-eye symptoms in contact lens wearers.

PURPOSE: To evaluate whether dry-eye symptoms are associated with epitheliopathy of that portion of the upper eyelid marginal conjunctiva-the lid wiper-that wipes the ocular, or contact lens surface, during blinking. METHODS: Subjects were divided into two groups based on the presence or absence of dry-eye symptoms. The lid wiper of asymptomatic (n=75) and symptomatic (n=30) soft contact lens wearers was examined, following the instillation of fluorescein and rose bengal dyes. Lid-wiper staining was graded zero to 3. RESULTS: Eighty percent of the symptomatic subjects displayed lid-wiper staining compared to 13% of the asymptomatic subjects. The difference in staining between the two groups was significant (P<0.0001). Of the symptomatic subjects, 20% showed no staining; 26.6%, grade 1 staining; 36.6%, grade 2; and 16.6% showed grade 3 staining. Of the asymptomatic subjects, 87% exhibited no staining; 9%, grade 1 staining; 3%, grade 2; and 1% showed grade 3 staining. CONCLUSIONS: This study describes a new clinical condition, lid-wiper epitheliopathy, an alteration of the epithelium of that portion of the marginal conjunctiva of the upper eyelid that wipes the ocular surface, diagnosed by staining with fluorescein and rose bengal dyes.

Adult↗

Chronic voluntary exercise may alter hepatobiliary clearance of endogenous and exogenous chemicals in rats.

A chronic voluntary exercise paradigm, which mimics the exercise pattern of many humans, influences the hepatic clearance of several organic anions and a bile acid, whereas a neutral organic compound is seemingly unaffected. To extend these observations, the present work has evaluated in female Sprague-Dawley rats the effect of 6 weeks of voluntary running on the hepatobiliary elimination of endogenous bile acids and glutathione and exogenously injected rose bengal, digoxin, and acetaminophen. Inactive rats had mobility limited to their cages, whereas exercised rats had free access to a 44-in running wheel. In comparison to weight-matched sedentary rats, the exercised rats ran 4.3 +/- 0.3 miles/day, consumed 45% more food daily, had slightly greater liver/body weight ratios, and slightly elevated basal bile flow rates. Biliary excretion of endogenous bile acids was increased significantly, and excretion of reduced and oxidized glutathione was increased in exercised rats by 190% and 173% of sedentary levels, respectively. Total clearance, biliary clearance, and maximal biliary excretion of the injected organic anion rose bengal (60 mumol/kg) were elevated in exercised rats by 86%, 440%, and 85%, respectively. In contrast, there were no observed differences in pharmacokinetic parameters, serum elimination, or biliary excretion for the clinically important cardiac glycoside digoxin (dose of 100 nmol/kg). Finally, study of the analgesic acetaminophen (330 mumol/kg) revealed that total and biliary clearances were increased by 37% and 42%, respectively, in exercised rats, whereas steady-state volume of distribution and elimination half-life were not significantly different.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetaminophen↗

Further evidence for an essential histidyl residue at the active site of pig liver 5-aminolevulinic acid dehydratase.

Photoxidation with methylene blue and rose bengal and chemical modification by diethylpryrocarbonate of pig liver 5-aminolevulinic acid dehydratase produced strong inactivation of the enzyme which was concentration dependent. Loss of enzyme activity by both photoxidation and ethoxyformylation was pH and time-dependent and protected by the presence of the substate and competitive inhibitors. The rate of inactivation was directly related to the state of protonation of histidyl groups, the unprotonated from being modified at a much faster rate than the protonated form. Plots of the pseudo-first order rate constants for 5-aminolevulinic acid dehydratase inactivation against pH resulted in typical titration curves showing inflection points at about pH 6.4 for methylene blue and rose bengal and 6.8 for diethylprocarbonate providing further and unequivocal evidence for the existence of critical histidyl groups at the active centre of the enzyme.

Animals↗

Selective media for detecting and enumerating foodborne yeasts.

No one medium is satisfactory for detecting, isolating and enumerating all yeasts in all foods. Antibiotic-supplemented media such as dichloran rose Bengal chloramphenicol agar, tryptone glucose yeast extract chloramphenicol agar, oxytetracycline glucose yeast extract agar and rose Bengal chloramphenicol agar are superior to acidified potato dextrose agar and other acidified media for enumeration of the vast majority of spoilage yeasts. Dichloran glycerol (18%) agar performs well for enumerating moderately xerotolerant yeasts. Malt extract yeast extract glucose (up to 60%) can be used for detecting and enumerating moderate and extreme xerophiles. These media also support the growth of moulds. Lysine agar, Schwarz differential agar and Lin's wild yeast differential agar are used by the brewing industry to differentiate wild yeasts from brewer's strains. Lysine agar is selective for apiculate yeasts and ethanol sulfite yeast extract agar is selective for Saccharomyces. Both have application in wineries. Modified molybdate agar can be used to selectively isolate yeasts from tropical fruits. Preservative-resistant yeasts can be detected on malt acetic agar. The recommended incubation temperature is 25 degrees C, but incubation time between plating and counting colonies ranges from 5 days for determination of general populations of yeasts to 10 days for more for xerotolerant yeasts. There is need for new and improved media for selectively isolating various groups, genera, species and strains of yeasts capable of growing only under specific environmental conditions in specific types of foods and beverages.

Bacteriological Techniques↗