Effects of Vitamin C and NADH on in Vitro Proliferative Function of Human Lymphocytes Exposed to Pesticides （Mancozeb and Metribuzin）

Antioxidants such as vitamin C and NADH can protect against pesticide adverse effects on immune function. The aim of this work was to study the in vitro effects of vitamin C and NADH on the proliferative responses of human lymphocytes exposed to pesticides （Mancozeb fungicide and Metribuzin herbicide） and on Thl and Th2 cytokine secretion. Their possible protective role on intracellular stress oxidative induced by pesticides was also investigated. Peripheral blood lymphocytes were isolated using differential centrifugation on a density gradient of Histopaque. They were cultured with mitogen concanavalin A （Con A）, Mancozeb and Metribuzin （50 μM） in the presence or the absence of vitamin C （50 μM） or NADH （50 μM）. Proliferation （MTT assay）, IL-2, INFγ and IL-4 （Elisa kits）, oxidative markers （intracellular superoxide anion, hydroperoxides, carbonyl proteins, GSH, catalase and SOD） were determined, The results showed that pesticides were immunosuppressive and decreased cytokine secretion with a shift away from to Thl phenotype. These immunomodulatory properties were accompanied by an increase in lymphocyte intracellular oxidative stress. The presence of vitamin C or NADH, in the medium, exhibited protective effects in human lymphocytes by inhibiting pesticide---induced lymphocyte proliferation suppression, inflammatory status and oxidative stress generation. In conclusion, vitamin C and NADH can result in a safe and effective method to reduce pesticide adverse effects and help to restore immune function.

Antioxidant antileishmanial cytotoxic and antimicrobial activities of a local plant Myrtus nivellei from Algeria Sahara

To study the phytochemical constituents and in vitro biological activities of hydromethanolic extract and fractions from Algerian Sahara Myrtus nivellei (M. nivellei) collected in Hoggar region and to identify the active fraction that can act as an alternative of commonly used antibiotics and as antileishmanial or antioxidant agents.MethodsPhytochemical screening of M. nivellei aerial parts was realised according to the literature. Extract was firstly prepared by using aqueous methanol then fractionated with ethyl acetate and butanol solvents. Total phenolics, tannis and flavonoids, of the hydromethanolic extract and their fractions were determined by Folin-Ciocalteu method as gallic acid equivalents and by aluminium chloride as rutin equivalent respectively. Extract and fractions were tested for their antimicrobial and antiparasital activities against standard bacteria using agar diffusion method and two kinds of leishmania visceral and cutaneous. The antioxidant activities were realized using phosphomolybdenum, FRAP and DPPH tests.ResultsPreliminary phytochemical screening exhibited the presence of flavonoids, tannins, saponins, and alkaloids. The experimental results showed that plant extract and fractions were high in phenolic compounds and exhibited an important role as antioxidant, antimicrobial and had a moderate antileishmanial activity.ConclusionsThese observations lead us toward more studies in this field, so that we can get more benefits from our local Algerian medicinal plants.

Synopsis of Fire＇s Forest in the Province of Sidi Bel Abbes： Analysis and Cartography

In order to characterize the forest fires in the province of Sidi Bel Abbes （North West of Algeria） and depending on a statistical data analysis of a 17-year period （1996-2012）, our works are oriented toward a study of the area covered by forest fires in the 53 constituent municipalities of the region. To do this, we depended on the archives of the forest fires balance sheets in forest conservation in the wilaya of Sidi Bel Abbes. The results show that the region lost an annual average 29,929 ha of its total forest area for a maximum of 11,817.5 ha recorded in 2012. On the other hand, the use of the fire severity index ＂FSI＂ by town and its cartography in which we used the map of administrative division of the study area as support, has divided the region into six classes according to the risk of fire, where the municipalities belonging to the forest of Telagh whose land is rugged represent the risk of fire is greater. Indeed, this situation category the study area among the most exposed wilaya and the most affected by this problem at the national level and require new approaches to forest management for its conservation. These are paramount in reducing the risk of fires based on appropriate management plans to be established for each forest conservation area.

Rapid increase in numbers and change of land-use in two expanding Columbidae species(Columba palumbus and Streptopelia decaocto)in Algeria

Background: Investigating population ecology of urban bird species,particularly the invasive and expending spe?cies,is the key for the success of urban management and planning strategies.Methods: Populations of two Columbidae species,the Woodpigeon(Columba palumbus) and the Eurasian Collared Dove(Streptopelia decaocto),were monitored from 1992 to 2010 in the Algiers Sahel,the Mitidja Plain and surround?ing areas(Northern Algeria).Monitoring of species densities was performed by the mapping?plot method.The foraging flight routes of Woodpigeon and the distribution of Eurasian Collared Dove were assessed by systematic observations.Results: The number of counted birds was statistically constant at first years of the survey,and then increased sig?nificantly,beginning from low numbers,their densities accelerated sharply between 2001 and 2006,then reached to an equilibrium state.The significant increase in Woodpigeon's population could be explained by the reduced hunt?ing pressure and by species adaptation to new food resources provided by nearby agriculture.Indeed,monitoring of flight directions of the species revealed the use of agricultural landscapes and habitats,which is a good indicator explaining adaptation and trophic niche of the species.The occurrence of the Eurasian Collared Dove in Algiers began in 2000.Its density experienced a rapid increase with similar trend pattern as that of Woodpigeon.Its distributional range is confined mainly in suburban environments of the Mitidja Plain.Conclusion: The modifications of habitats,urbanization increase and the lessening of hunting in the Mitidja Plain facilitated the rapid expansion of the Eurasian Collared Dove and Woodpigeon as well contributed to the increase in their numbers over time.

Evaluation of Anti-Hyperglycemic and Anti-Hyperlipidemic Activities of Water Kefir as Probiotic on Streptozotocin-Induced Diabetic Wistar Rats

Diabetes mellitus is a predominant chronic disease which causes mortality of millions of people yearly. Its prevalence is on the rise worldwide. Water kefir is fermented food produced by a matrix of polysaccharides containing bacteria and yeasts, with therapeutic properties. Our study aimed to evaluate anti-hyperglycemic and anti-hyperlipidemic activities of water kefir on streptozotocin-induced diabetic Wistar rats. Adult Wistar rats were made diabetic by intraperitoneal injection of streptozotocin, and were given or not kefir in drinking water for 5 weeks. Body weight, glucose and lipid levels were measured. The results demonstrated evident improvement in body weight, glucose, and lipid profiles of treated rats comparing with diabetic or control rats. Water kefir is found to be less cost hypoglycemic and hypolipidimic treatment and less time consuming. Water kefir can potentially be useful food for diabetes to control glucose and lipid levels.

Blood Oxidant/Antioxidant Status in overfed Aging Rats: Comparative Effects of Sunflower and Olive-Linseed Mixture Oil Rich Diets

Obesity, one of the most common health problems, is associated with glucose and lipid metabolism abnormalities, increased cardiovascular risk and oxidative stressI1}. Aging is also associated with an increase in oxidative stress[2]. Considering that older adults are already at risk for oxidant stress, the additional metabolic perturbations of obesity aggravated the susceptibility to oxidant stress. Modulating age-related oxidative stress via dietary interventions could reduce free radical production and significantly improve health in aging.

Nutritional and Metabolic Profile in Diabetic Patients and Relationship with Metabolic Syndrome

Background & Objectives: Diabetes and metabolic syndrome spread alarmingly throughout the whole world including Algeria, so our study makes the links closer between these two entities and that through studying nutritional, metabolic and physical profiles. Study Design: 204 patients were recruited and interviewed (anthropometric measurements, blood pressure, age, sex, personal and family history, the practice of physical activity and evaluation of food consumption). Patients and Methods: A cross-sectional survey was conducted in the western Algerian region (Sidi Bel Abbes). Patients with MetS are defined according to the criteria of the National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III). Results: The results show that the frequency of the metabolic syndrome is 86.27% with predominance of women (92.15% women vs 80.39% men). The age group most affected by the MetS among women and men is [54 - 79] years. We also note that 51.28% of men present three criteria of MetS, while 65.21% of women have four and five criteria. The results show that the criteria most dominant in men are hypertension and type 2 diabetes, whereas in women, the high waist is the most abundant criterion. Moreover, the whole is underlined by a low physical activity with only 21.56% who practice it regularly (15.95% women vs 28.04% men). The estimate of food intake shows a qualitative imbalance: protein intake is 19.65% in women vs. 19.43% in men represented mainly by vegetable protein (83.72% for women vs. 72.85% for men);lipids intake is characterized by a lower consumption than the recommendation of the Mediterranean diet concerning the mono-unsaturated fatty acids (MUFA) (39.44% among women vs 40.24% at the men), as in poly unsaturated fatty-acids including (PUFA) (23.30% among women vs 23.64% at the men) whereas an increase in the consumption of the saturated fatty-acids (SFA) is observed in the whole population including (37.24% among women vs 36.10% at the men);lower concentration in calcium, magnesium and en fibers;important concentration of sodium and an insufficient contribution of water. Conclusion: We should be aware of the importance of the modulation of these risk factors through harmonization of “lifestyle” to prevent the occurrence of metabolic syndrome.

Qualitative and Environmental Aspects Study of Water Distributed in West Algeria： The Case of Sidi Bel Abbes City

Water comes from various different sources, such as lakes, rivers, streams, springs and boreholes. Depending on the quality of its source, water will almost certainly have to be treated to make it safe to drink. All water supplies for human consumption must be well stored, free from micro-organisms, parasites and any substances that endanger public health if found in sufficient numbers or concentrations, in the same time. In our continued effort for a better result in our study of qualitative aspects, by physicochemical and bacteriological analysis, the authors focus in two crucial questions： a） is the water quality of Sidi-bel-Abbes good or bad ,and well stored？ b） is the people of Sidi-bel-Abbes are aware of the importance of water and knowledgeable about how to use this vital？ This study is the first to use data from I real work based upon a developed method and a good investigation to know exactly if water of Sidi-Bel-Abbes is safely treated, well stored and consumed in resemble way. In first, the authors plotted the method in analysing the qualitative aspect of water using developed material for a precise result starting by the distribution connection （condition and management） and the verification of water tank storage and finishing by analyzing the all the aspect of water （physicochemical and bacteriological analysis）. In second, the authors focus on the way that people of Sidi Bel Abbes used and consumption water by introducing a targeted method （questionnaires） which help us a lot to know precisely the way, time and the real quantity of water consumed daily. In final step of our study of qualitative aspects, by physico-chemical and bacteriological, and environmental analyses, investigation processed by PCA （principal component analysis） of the drinking water supplied to Sidi Bel Abbes city （Algeria north-west） concluded by an encouraging result, that despite the various constraints faced by the departments concerned, they ensure the supply of safe drinking water in households almost daily.

Vitamins C, E, and NADH on in Vitro Lymphocyte Proliferation and Redox Status among Obese Patients

Background: The prevalence of overweight and obesity associated with oxidative stress and immune abnormalities is continuously increasing. Antioxidant supplementations might counteract potential damage caused by ROS to cellular tissues. Objective: To determine the role of vitamins on immune improvement during obesity, we investigated in vitro effects of vitamins C, E, and NADH on mitogen-stimulated proliferation, Th1- and Th2-type cytokine production, and oxidant/antioxidant status of lymphocytes isolated from obese patients. Methods: Peripheral blood lymphocytes were isolated using a density gradient of Histopaque. They were in vitro cultured and stimulated by Con A in the presence or absence of vitamins. Cell proliferation was determined by MTT assay and interleukin-2, interleukin-4 and interferon-γ (INFγ) secretions. Cell oxidant/antioxidant balance was studied by assaying glutathione (GSH), malondialdehyde (MDA), carbonyl protein levels, catalase activity and micronucli frequency. Results: Obesity is associated with enhanced oxidative stress response. Indeed, vitamin C, E and NADH improved significantly lymphocyte proliferation and diminished cellular oxidative stress. Conclusion: Treatments of lymphocytes with vitamins had beneficial effects on lymphocyte proliferation, cytokines secretions and redox status, generating an anti-inflammatory profile and should be considered in therapeutic approaches for normalizing immune cell function in obesity.

Situation of the Technical and Financial Assistance According to Farms Opinions in Some Mountainous Regions of Tlemcen Province (Algeria)

The technical and financial assistance plays a fundamental role in agriculture in order to maintain its durability especially in disadvantaged regions. For this purpose, the present work aims to evaluate the technical and financial assistance according to the opinions of farms in some mountainous areas belonging to the mountains of Tlemcen wihch are situated in the north-western Algeria. The results obtained showed that to improve the relation between bank and farms, public financial institutions should review the conditions for bank credits granting according to sociological particularity of the Algerian population who is very attached to the instructions of Islam religion which prohibits interest rates generated by bank credits. In addition, strengthening these mountainous areas by qualified technical personnel is a necessary operation to improving the knowledge and the know-how of farmers which are very insufficient in these areas.

Contribution to the Physicochemical and Organoleptic Study of Two Olive Oils of Traditional and Industrial Extraction of the Wilaya of Tlemcen

Virgin olive oil is obtained from the fruit of the olive tree, and only using physical processes, without resorting to refining steps, the absence of the latter allows the olive oil to retain all its antioxidants. This work is a contribution to the characterization of the properties of olive oil in the region of Tlemcen. They relate more particularly to the comparison of the physicochemical properties of two oils, one from traditional extraction and the other from industrial extraction. To do this, the first objective of studies of the two chains of transformation (traditional and industrial) was made followed by a physicochemical and organoleptic comparison of these two oils of olives, to conclude on the advantages and disadvantages of each of the two processes.

Phytoecological and phytoedaphological characterization of steppe plant communities in the south of Tlemcen(western Algeria)

In Algeria, the steppe areas of southern Sebdou between Tlemcen, El-Aricha and Mecheria are the scene of an adverse and continuous ecological imbalance often caused by the strong support of human pressure in these ecosystems. In arid and semi-arid regions, salinity remains a constraint for the development of plants. This study focuses on the realization of floristic surveys, where attention was paid to areas occupied by the dominant perennial species (Tamarix gallicaL.). A correspondence analysis by Minitab 15 software has allowed us to individualize the groups of species attracted by some parameters (edaphic, nitrates, humidity), and ecological gradients appear to affect the distribution of these taxa. The study of plant diversity shows the dominance of biological type of therophytes (41%) and morphological type of the Chenopodiaceaes with (25%). For geographical types, the strict Mediterranean and circum-Mediterranean element predominates.

Optimization of Extraction Conditions of Some Phenolic Compounds from White Horehound (<i>Marrubium vulgare</i>L.) Leaves

This research was aimed to optimize the extraction conditions of three phenolic compounds: total phenolics, flavonoids and condensed tannins, from White Horehound’s leaves (Marrubium vulgare L.). Distilled water and different organic solvents such as: methanol, ethanol and acetone, were used, with various concentrations (20% - 80%, v/v), temperatures (20&deg;C - 60&deg;C) and extraction times (30 - 450 min). Results showed that the maximum total phenolics amounts (293.34 ± 14.60 mg gallic acid equivalent/g dry weigh), were obtained with 60% aqueous methanol at 25&deg;C for 180 min;total flavonoids (79.52 ± 0.55 mg catechin equivalent/g dry weigh) with 80% aqueous methanol at 20&deg;C for 450 min, and condensed tannins (28.15 ± 0.80 mg catechin equivalent/g dry weigh) with 60% aqueous acetone at 50&deg;C and for 180 min. ANOVA test showed the significant effect (***P < 0.001) of the extraction conditions tested on phenolic compounds. The Principal Component Analysis (PCA) exhibited the positive effect of low temperatures on total phenolics and flavonoids extraction, and the effect of high temperatures on the condensed tannins extraction. The Response Surface Methodology (RSM) provided predicted values of extraction conditions and maximum polyphenols amounts similar to those obtained experimentally.

Factors Facilitating and Forcing the Breast-Feeding to Tebessa (East City Algerian)

To estimate the situation of the breast-feeding, we made a survey (investigation) in the wilaya of Tébessa in Algeria. Our sample was chosen at random. He is constituted by 200 mothers and 200 children aged 0 to-24-month. A questionnaire, beforehand established, is subjected to the mothers. Our statistical analysis is based on the test of Chi2 and the calculation of the percentages. It emerges from our work that: the exclusive breast-feeding is practised by 50.50 % of the cases. The breast-feeding is less practised by the salaried women 40.91 %. The practice of the exclusive feeding increases when the academic level of the mother decreases (50.80% vs. 46.15%), the rate of the mixed feeding and the artificial feeding increases when the academic level increases. The circle of acquaintances plays an important role in the choice of the mode of feeding with 53.47% of the cases. The first food given to the Child within first hour which has followed the childbirth: the maternal milk with 66.5% was. The main causes of the stop of the breast-feeding are: the health of the woman, the incapacity of the milk, the resumption of work the aesthetics of the woman and the arien of a new pregnancy. 72.78%, of the mothers have chooses the breast-feeding because it is better for the health of the child. The average duration of the feeding is of (4.68 ± 4.437 months) the day of the survey. A better information of the population about the feeding and a rationalization of the marketing of industrial milk, could help in a better promotion of the breast-feeding.

<i>In Vitro </i>Effects of Oil’s Fatty Acids on T-Cell Function of Obese Men

Background: Dietary fatty acids have important homeostatic functions in regulating the immune response and may exert beneficial effects on immune alterations during obesity. Objective: To assess the in vitro effects of oil fatty acids, different oils (olive, linseed, Nigel, sunflower) were tested on T-lymphocyte proliferation, Th1- and Th2-type cytokine production, and intracellular oxidant/antioxidant status in obese patients. Methods: Peripheral blood lymphocytes were isolated using Histopaque and were in vitro cultured and stimulated by Con A in the presence or absence of the oils. Cell proliferation, interleukin-2, interleukin-4 and interferon-γ (INFγ) secretions and intracellular oxidative status (glutathione (GSH), malondialdehyde (MDA), carbonyl protein levels, catalase activity and micronuclei frequency) were investigated. Results: Abnormalities in lymphocyte function and intracellular oxidative stress were observed in obesity. Linseed oil induced a reduction in T-lymphocyte proliferation and IL-2 production while Nigel oil increased them in both obese and control groups. In addition, Nigel oil enhanced IFNγ and IL-4 secretion. Olive and sunflower oils had no effect on lymphocyte proliferation and cytokine secretion in both groups. Linseed and Nigel oils induced an increase in T cell GSH concentrations and catalase activity with a concomitant decrease in MDA, carbonyl protein contents and micronuclei frequency especially in obese patients. Conclusion: Linseed and Nigel oils had beneficial effects on lymphocyte proliferation, cytokines secretions and redox status, while olive and sunflower oils had no effects on immune cell function in obesity.

Effects of Maternal Linseed Oil Supplementation on Metabolic Parameters in Cafeteria Diet-induced Obese Rats

Because linseed oil may influence maternal and fetal metabolisms, we investigated its role in the modulation of lipid metabolism in cafeteria diet-induced obese rats and their offspring. Female Wistar rats were fed control or cafeteria food, which were either supplemented or not supplemented with linseed oil （5%） for I month before and during gestation. At parturition, serum and tissue lipids and enzyme activities were analyzed. Cafeteria diet induced adverse metabolic alterations in both mothers and offspring. Linseed oil improved metabolic status. In conclusion, linseed oil displayed health benefits by modulating tissue enzyme activities in both obese mothers and their newborns.

In vitro assessment of antioxidant,neuroprotective,anti-urease and anti-tyrosinase capacities of Tamarix africana leaves extracts

OBJECTIVE:To characterize the chemical profile of methanolic crude extract and its fractions(Ethyl acetate,n-butanol and aqueous)using liquid chromatographymass spectrometry(LC-MS)analysis,to evaluate their biological and pharmacological properties:antioxidant(1,1-diphenyl-2-pycrylhydrazyl(DPPH),2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic)(ABTS·+),galvinoxyle free radical scavenging,reducing power,phenanthroline andβcarotene-linoleic acid bleaching assays),enzymes inhibitory ability against several enzymes[acetylcholinesterase(AChE),buthyrylcholinesterase(BChE),urease and tyrosinase].METHODS:Secondary metabolites were extracted from Tamarix africana air-dried powdered leaves by maceration,the crude extract was fractionated using different solvents with different polarities(Ethyl acetate,n-butanol and aqueous).The amount of polyphenols,flavonoids and tannins(hydrolysable and condensed)were determined using colorimetric assays.A variety of in vitro biochemical tests were carried out to assess antioxidant and oxygen radical scavenging properties using DPPH,ABTS,galvinoxyle free radical scavenging,reducing power,phenanthroline andβcarotene-linoleic acid bleaching methods.Neuroprotective effect was examined against acetylcholinesterase and buthyrylcholinesterase enzymes.The anti-urease and antityrosinase activities were performed against urease and tyrosinase enzymes respectively.The extract’s components were identified using LC-MS and compared to reference substances.RESULTS:The results indicated that Tamarix africana extracts presented a powerful antioxidant activity in all assays and exhibited a potent inhibitory effect against AChE and BChE as well as urease and tyrosinase enzymes.LC-MS analysis identified amount of eight phenolic compounds were revealed in this analysis;Apigenin,Diosmin,Quercetin,Quercetine-3-glycoside,Apigenin 7-O glycoside,Rutin,Neohesperidin and Wogonin in methanolic extract and its different fractions of Tamarix africana from leaves.CONCLUSIONS:Based on these findings,it is reasonable to assume that Tamarix africana could be considered as a potential candidate for pharmaceutical,cosmetics,and food industries to create innovative health-promoting drugs.

Effect of a normal protein diet on oxidative stress and organ damage in malnourished rats

BACKGROUND： We investigated the effects of three weeks of renutrition with a normal protein diet on oxidant/antioxidant status in malnourished rats using biochemistry and histology. METHODS： Eighteen young Wistar rats were divided into three groups： control group was fed on a normal protein diet; malnourished group was fed on low protein diet and renourished group was fed on low protein diet followed by a normal protein diet. Serum albumin was evaluated. Malondialdehyde, protein carbonyl, superoxide dismutase and catalase levels were determined in the intestine, muscle and liver. Intestinal and hepatic damage were assessed by histological examination. RESULTS： Protein malnutrition resulted in a significant decrease of body weight, albumin level, villus length, intraepithelial lymphocytes counts （IELC） and superoxide dismutase level （liver and muscle）. However, catalase activity increased significantly in muscle and gut but there was no difference in liver. In all organs, malondialdehyde and protein carbonyl content of malnourished group showed a significant increase. Interestingly, a normal protein diet for three weeks resulted in a return to normal levels of superoxide dismutase, albumin, malondialdehyde and protein carbonyl in all organs. Catalase activity decreased in the muscle and gut and exhibited no significant difference in the liver. The renutrition diet enhanced also the recovery of intestinal epithelium by increasing villus length. Hepatic damage of rats fed normal protein diet was markedly reduced （macrovesicular steatosis decreased by 45%）. CONCLUSION： The normal protein diet could improve the oxidant/antioxidant imbalance and organ damage induced by protein malnutrition.

Report on the Diatoms and Dinoflagellates Distribution along the Algerian Coasts: Inter-Region Comparison

The phytoplankton group’s composition is an excellent indicator of the state of coastal waters. The aim of this study is to explore spatial variability of phytoplankton in shallow coastal areas from different regions(East, Center and West) of the Algerian coast. The quantitative and qualitative study of the phytoplankton population was conducted on samples taken during the 2012–2013 period by the research vessel GRINE BELKACEM. The qualitative results show a significant diversity of diatoms and dinoflagellates, which rank first and second in the total flora(Fr > 50%). This finding is also confirmed quantitatively by the abundance values in both 2013 and 2012 samples which largely exceeds the average value in most other areas of the western region(56%) and slightly exceeding 52% in all central areas which represent an equi-distribution between diatoms and dinoflagellates. The Dinoflagellates contribute significantly to the quantitative richness in the eastern region(>60%). The minimum value of Dia/Dino index, recorded in the eastern region confirms the dominance of the dinoflagellates especially in Skikda(0.31)(Skikda).However, our results reveal the presence, in smaller proportion, of other groups such as the cocolithophoridae and euglenophyceae.

Variations in Disease Resistance of Chestnut Cultivars against Chestnut Blight Fungus,Cryphonectria parasitica

Chestnut blight caused by Cryphonectria parasitica is one of the major chestnut diseases occurred on stems and branches. Oriental chestnut has more resistance than American one. Twenty-five cultivars of chestnut treeincluding various cultivars such as Tsukuba, Okkwang, Rihei, Ginyose, Eunsan, etc. were tested with a rapid, simple and reliable method. The cultivar Banseki is one of the most resistant cultivars tested and has the necrotic area of 0.40 cm2. However, the necrotic areas of the cultivars of American chestnut, Eunsan, Kwangjujoyul and Eaton, ranges from 3.35 to 2.61 cm2 and these cultivars are relatively susceptible to this pathogen. Many pycnidia of this pathogen were observed on the inner and outer bark tissues after incubating for 3 weeks.