Running Tap Water Research Articles

First Report of Diaporthe amygdali Associated with Twig Canker and Shoot Blight of Nectarine in Spain.

Nectarine (Prunus persica (L.) Batsch var. nucipersica (Suckow) C. K. Schneid.) is a fruit crop widely cultivated throughout the Mediterranean basin. In Spain, it is mainly grown in eastern regions of the country. In March 2018, 5-year-old nectarine trees showing twig canker symptoms were observed after a rainy spring period in a 0.5 ha orchard located at Alaior, Menorca island (Spain). Cankers were frequent on affected trees (approximately, 80% of the total trees), thus leading to shoot blight. Ten twig segments of one-year old wood with cankers were cut, washed under running tap water, surface disinfected for 1 min in a 1.5% sodium hypochlorite solution and rinsed twice in sterile distilled water. Small pieces (2 mm) of affected tissues were taken from the margin of the cankers and plated on potato dextrose agar (PDA) supplemented with 0.5 g/L of streptomycin sulphate (PDAS). The plates were then incubated at 25 ºC in the dark for 7 to 10 d. Actively growing colonies were first hyphal-tipped and then transferred to PDA and 2% water agar supplemented with sterile pine needles and incubated at 21-22ºC under a 12h/12h near UV / darkness cycle during 21 d (León et al. 2020). Colonies were white at first, becoming light cream, with visible solitary and aggregate pycnidia at maturity. Alpha conidia were aseptate, fusiform, hyaline, multi-guttulated (mean ± SD = 7.4 ± 0.7 × 2.8 ± 0.4 µm, n = 100). Beta and gamma conidia were not observed. The morphological and cultural characteristics of the isolates were congruent with those of Diaporthe spp. (Gomes et al. 2013). The ITS1-5.8S-ITS2 (ITS) region and fragments of β-tubulin (tub2), the translation elongation factor 1-alpha (tef1-α) gene regions, histone H3 (his3) and calmodulin (cal) genes of representative isolate DAL-59 were amplified and sequenced (Santos et al. 2017). The BLASTn analysis revealed 100% similarity with sequences of D. mediterranea (Synonym D. amygdali) (Hilário et al. 2021) isolate DAL-34 from almond (ITS: MT007489, tub2: MT006686, tef1-α: MT006989, his3: MT007095 and cal: MT006761). Sequences of isolate DAL-59 were deposited in GenBank Database (ITS: MT007491, tub2: MT006688, tef1-α: MT006991, his3: MT007097 and cal: MT006763). Pathogenicity tests were conducted using one-year-old potted plants of nectarine cv. Boreal, which were inoculated with isolate DAL-59. In each plant, a 3 mm wound was made in the center of the main branch (about 30 cm length) with a scalpel. Colonized agar plugs with 3 mm diameter, which were obtained from active 10-day-old colonies growing on PDA, were inserted underneath the epidermis and the wounds sealed with Parafilm. Inoculated plants were incubated in a growth chamber at 23 ºC with 12 h of light per day. Controls were inoculated with uncolonized PDA plugs. There were twelve plants per treatment, which were arranged in a completely randomized design. Five days after inoculation necrosis development was observed in the area of inoculation. Wilting and twig blight symptoms over the lesion occurred 3-wk after inoculation and pycnidia were detected, while the controls remained asymptomatic. Diaporthe amygdali was re-isolated from symptomatic tissues and identified as described above to satisfy Koch's postulates. To our knowledge, this is the first report of D. amygdali causing twig canker and shoot blight disease on nectarine in Spain.

Sensory Quality Evaluation of Superheated Steam-Treated Chicken Leg and Breast Meats with a Combination of Marination and Hot Smoking.

As the sensory qualities of meat processed using methods such as superheated steam, marination, and hot smoking have not been examined, this study analyzed the sensory quality of chicken meats (leg, breast) and its chemical correlation by determining optimal processing conditions (superheated steam treatment, marination, and hot smoking). Chicken meats were defrosted using room temperature, running tap water, or high-frequency defroster. Marinated meats with herbal extract solution were treated with superheated steam and then hot smoked with wood sawdust; sensory evaluations were performed at each processing step. The products were analyzed for fatty acids and nutrients, along with storage tests under different conditions. High-frequency defrosting showed the lowest drip loss and thawing time compared to other methods. Bay leaves and oak wood were selected as the best sub-materials for higher sensory scores. Optimal superheated steam conditions showed higher overall acceptance (8.86, 8.71) and were set as follows; leg meat (225 °C; 12 min 20 s), breast meat (223 °C; 8 min 40 s). The final meat products possessed good nutritional composition and no severe sensory spoilages were detected during storage despite microbial and chemical degradations. Thus, regular sensory evaluations at each processing step and storage condition were effective for developing superior chicken meat products.

The use of different detergents in skeletal preparations

Skeletal preparation has become an integral component within the field of forensic anthropology. The aim of this study was to determine which commercial detergent was most effective and efficient for use in skeletal preparation. The hind limbs of 24 pigs (Sus scrofa) and five detergents with bleaching agents and enzymes (Surf and Ariel), only enzymes (OMO Auto and Sunlight powder) or only bleaching agents (Sunlight dishwashing liquid) were used. Specimens were skinned and immersed into a pre-heated 6 L detergent solution or tap water and macerated at either 45 °C, 50 °C, 55 °C and 60 °C. When maceration was deemed complete any remaining soft tissue was manually removed under running tap water and the remains left to dry. A scoring system was utilized to assess the effectiveness and efficiency of each detergent. OMO Auto specimens only required a single day to complete macerate regardless of the temperature and these specimens constantly scored better than the other detergents used, thus making it the most effective and efficient detergent tested.

Evaluation of biofilm-forming abilities of Listeria monocytogenes (ATCC 19115) and efficacy of different washing methods for removal of biofilm on apple

Fresh produce can be contaminated at any stage along the food supply chain. In this study, apple was chosen to determine the time course of biofilm formation by Listeria monocytogenes (ATCC 19115), as well as to compare the efficacy of different household washing methods such as scrubbing with hands under running tap water, soaking with and without commercial vegetable wash with different treatment times in removing the biofilm formation by L. monocytogenes on apple surface. The biofilm formation was quantified using crystal violet assay and the result showed that L. monocytogenes took 18 hrs to form matured biofilm on apple surface. Besides, scrubbing apples with hands under running tap water for 30 s and 60 s were the most effective method which significantly removed (P<0.05) biofilm formed on the apple surface with approximately 5.93 log reduction. Soaking apples with vegetable wash for 5 mins and 10 mins were also found to be significantly effective (P<0.05) in reducing L. monocytogenes biofilm. Since L. monocytogenes can form matured biofilm on fresh produce, therefore efficient washing step is important before consuming fresh produce to lower the risk of foodborne illness.

EVALUATION OF SURFACE QUALITY AND DIMENSIONAL ACCURACY OF ELASTOMERIC IMPRESSION MATERIALS AFTER VARIOUS DISINFECTION PROTOCOLS

To evaluate surface quality and dimensional accuracy of elastomeric impression materials after various disinfection protocols. A Total of 150 samples were made i.e..50 from each type of impression material. Specimens were immersed in disinfectant solutions namely 0.2% chlorhexidine,5.25% hypo,0.2% povidone iodine, running tap water and ozone water for disinfection and examined for surface quality using stereomicroscope and dimensional accuracy using digital Vernier calipers. Silicones exhibited best dimensional accuracy & surface quality when treated with ozone water after 24 hours of disinfection whereas polyether exhibited good surface quality & dimensional accuracy when treated with control (tap water) after 24 hours of disinfection.

PM 7/121 (2) ‘Candidatus Liberibacter africanus’, ‘Candidatus Liberibacter americanus’ and ‘Candidatus Liberibacter asiaticus’

Specific scopeThis Standard describes a diagnostic protocol for ‘Candidatus Liberibacter africanus’, ‘Candidatus Liberibacter americanus’ and ‘Candidatus Liberibacter asiaticus’.1This Standard should be used in conjunction with PM 7/76 (5) Use of EPPO diagnostic protocols.Specific approval and amendmentFirst approved in 2014–09. Revised in 2021–04.

Characteristics and methods to release seed dormancy of two ground cherry (Physalis) species

Several ground cherry (Physalis) species have attracted significant interest due to their potential use in medicines. Nonetheless, these species are invasive weeds in some regions of the world, while cultivated for food and medicinal uses in the others. Physalis angulata is cultivated for its medicinal potential, whereas P. philadelphica is grown as a horticultural crop. Both species are regarded as invasive weeds in Turkey causing significant economic losses. These species are underutilized in the country and could serve as a valuable resource for agriculture and medicine industries. Therefore, we investigated the seed dormancy type, levels and methods of dormancy release for P. angulata and P. philadelphica. Seeds of different populations of both species (3 population per species) were collected from various elevation gradients in southeastern Anatolia region, Turkey. Numerous seed dormancy-release techniques were evaluated to understand the requirements for dormancy release. Significant differences were observed in seed dormancy level among species and their populations, while viability remained similar. Seeds P. angulata had higher dormancy level (73.07–98.78 % dormancy) than P. philadelphica (7.56–33.15 % dormancy). The level of seed dormancy was increased with increasing elevation gradient in both species. There were some germination inhibiting substances, which retarded seed germination of both species. This indicated that the dormancy type in both species belongs to physiological dormancy (PD). Running tap water proved the most effective treatment in releasing seed dormancy of all tested populations. Some of the dormancy breaking techniques, especially scarification with H2SO4, even suppressed germination. The knowledge of seed dormancy could be utilized to commercialize/cultivate both species in Turkey. Nevertheless, the result could also be used for the development of management strategies against the species where these are considered as invasive weeds.

아연 이온화 장치의 상수 중 철 시편과 수도 배관에 미치는 부식억제 효과 예측

This study quantitatively investigated the inhibition effect of corrosion on iron coupons and water pipes by ionization of zinc metal in tap water line. After installing iron coupons into water pipes and running tap water for 20 days, the average difference of the annual corrosion rates calculated from the weight losses of iron coupons with or without the ionization device was estimated to be 12.8 %. Comparing corrosion rates with or without corrosion inhibitor on new iron coupons showed higher difference in corrosion rates, but over time, the difference of corrosion rates between absence and presence of inhibitor had decreased gradually. It was also confirmed that the less rust-free pure iron area on the iron specimen and the less pure iron surface to react with oxygen in water gradually reduces the corrosion rate. X-ray diffraction analysis of the rust generated on iron coupons confirmed that most of them were red hematite(Fe₂O₃), and that some of the blackish parts were magnetite. It is known that the rust generated causes red tap water to fall, some of which develop into foot corrosion, resulting in pin-holes in the water pipes. Therefore, based on existing research results, we predicted the breaking time due to foot corrosion of iron coupons with or without corrosion inhibitors. Zinc corrosion suppressors were installed in water pipes where corrosion had already progressed and rusted, and the internal surface of the water pipes was observed to increase the relative ratio of the black magnetite components over time, and it was believed that corrosion suppressors could be expected for a long time.

A STUDY OF THE MORPHOLOGY OF PLACENTA IN TERTIARY CARE HOSPITAL OF CENTRAL KARNATAKA, INDIA

Introduction: Placenta is a temporary structure, developed during Pregnancy and discarded at Parturition. Human Placenta is of Discoid and Haemo-Chorial type. It connects the foetus with the mother's womb. It is formed by the contribution of two Individuals - both Mother and Baby. At 1 term four- fth of Placenta is of foetal origin and one- fth is of maternal. Along with Amnion, Chorion, Allantois and Yolk sac, Placenta can also be considered as one of the foetal membranes, which acts as Nutritive and Protective to the developing Embryo. Method: - The present study was done on morphology and morphometry of 150 placentae carried at the Department of Anatomy, Basaveshwara Medical College & Hospital (BMCH), Chitradurga. The placentae were collected with prior ethical clearance and proper consent. Soon after the delivery the placental surface was washed thoroughly under running tap water which were then weighed, tagged and brought to the department of Anatomy, BMCH. The parameters like size, shape and attachment of umbilical cord were noted down. Gross examination was carried out according to the proforma. Result: - The maximum number of placentae belong to the range of 501-600 gms (38.7%). Least placental weight is recorded as 205 gms, showing dispersal pattern. The maximum recorded weight is 835 gms and the average being 491.4 gms. The majority of the placentae are discoid (63.3%). The minimum length recorded is 8.9 cms, maximum as 23.80 cms and the average being 18.3 cms. Conclusion: - Careful examination of Placenta can help in explaining adverse outcomes, their management in subsequent pregnancies and assessment of new born risk.

Alkaline Treatment on the Chemical and Morphological Properties of Kelempayan (<i>Neolamarckia cadamba</i>) Wood

Alkaline treatment has a function to disrupt hydrogen bonding in network structure and to remove some hemicellulose, lignin and extractives for efficient contact with the matrix when preparing composites. Kelempayan (Neolamarkiacadamba) particles were treated using 0.5, 1 and 2% sodium hydroxide (NaOH) solution at a temperature of 90 °C for 60 minutes periods of immersion. The particles were washed with running tap water to neutralize excess NaOH and oven dried at 80 °C. The treated particles were then grounded to a fine size before being sieved to 60 mesh size of sawdust. Finally, the treated samples were air dried before chemical analysis. The sampling and preparation of sawdust were carried out according to the TAPPI Standard T257 cm - 02 (2002). The changes occurring on Kelempayan’s surface were assessed using Scanning Electron Microscopy (SEM). Chemical analysis indicated that hemicellulose, lignin and extractives became decreased by increasing alkaline concentration. Morphological analysis showed that treated surface was cleaner and rougher than that of untreated.

CATTLE SLAUGHTERING AND POSTMORTEM HANDLING PRACTICES IN SELECTED NIGERIA CITIES

Cattle slaughtering and postmortem handing practices in Nigerian abattoris and markets were investigated. The study revaled poor pre- and post slaughter handling of cattle and carcass. The abattoirs were inadequately were inadequately equipped and maintained. The abbattors were characterized by lack of running tap water and proper waste disposal facilities. A critical assessment of the slaughtering process in the abattoirs when compared with the standard method showed a total absence of stunning (necessary for humance slaughtering) and aging or ripening (Which enhances tenderness and juiciness of meat) unit operations. The beef marketing chain was also found to be poorly organized and managed with associated poor preservation and storage facilities. Suggestions for improvement were also discussed.

The Effect of Storage andWater Rising of Microbial Contamination of Carrots and Cucumbers.

In this work the Effect of storage and water rinsing in reducing the microbial contamination of carrots and Cucumbers and the time of spoilage was evaluated. In storage experiment done , washed and unwashed of carrots and cucumbers samples were stored at room and refrigerator temperatures.Washing with running tap water resulted in big reduction of the microbial load of carrots and cucumbers. The reduction ranged between 85 – 99 % for the total bacterial count, 92 % for moulds, and between 74 and 90 % for coliform bacteria. Unwashed samples and samples stored at room temperature spoiled more quickly than the washed samples and the sample stored in the refrigerator. In general carrots samples stored at room temperature spoiled in six to seven days and those store in the refrigerator spoiled in nine to ten days ,with respect to cucumbers spoilage occurred often six days at room temperature and after seven days in the refrigerator.From this experiment we found that carrots was more resistant to spoilage than the cucumbers. Carrots and cucumbers spoilage was eminent when the microbial load reached about 106 ????????????/????.It was concluded that, although the thorough cleaning of salad Carrots and Cucumbers with water and their storage in refrigerator would greatly reduce their microbial load, elimination of all microbial would not be achieved.

In Vitro Clonal Propagation of Nardostachys jatamansi: A Traditional Himalayan Medicinal Plant

Plant tissue culture method has an impressive technique for Investigation and Explains basic and applied problems in plant biotechnology field. Micropropagation has played a vital role in the rapid multiplication of many plants species. The nodal explants and shoot tip of N. jatamansi inoculated in MS medium (Murashige and Skoog) contain different types concentrations of PGRs (Phytohormones) at various frequencies for the optimization of growth quality for shoot bud Induction, shoot proliferation and micro rooting in plant. The perfect shoot induction takes place in the concentration of BAP + IBA (2.0 mg/l +1.5 mg/1) multiplication of nodal explants and shoot tip in the combination lower concentration of BAP and KN (2.0 mg/1+1.5 mg/1) This combination proved best for multiple shoot formation. Half strength (1/2) of the MS medium containing NAA and BAP (1.5 mg/1+1.0 mg/1) in combination was most useful for rooting in plant. Well developed rooted micro shoots were smoothly removed for the culture flask and dipped in 70% ethanol for 2 min and then washed with running tap water for 5-10 min to remove media for the root and transferred to small plastic cups carry cocopeat, garden soil and sand (2:2:1) and produce healthy growth in ex-vitro conditions.

Cross-contamination events of Campylobacter spp. in domestic kitchens associated with consumer handling practices of raw poultry

Contaminated poultry is the major vehicle for consumer's exposure to Campylobacter. This study aimed to perceive potential cross-contamination events during preparation of raw poultry that can contribute to the spread of Campylobacter spp. in domestic kitchen environments and to understand consumers' meanings and justifications on preparation of a poultry dish at home. A total of 18 households were visited to observe consumers preparing a recipe that included poultry. Poultry samples and swabs from the kitchen surfaces and utensils, such as kitchen cloth, hand towel, sponge, cutting boards and the sink, were collected before and after food preparation and tested for the presence of Campylobacter spp. Genotypic characterization of 72 Campylobacter spp. isolates was carried out through Pulse-Field Gel Electrophoresis (PFGE). Fourteen chicken samples were contaminated with Campylobacter spp. (77.8%). Twelve consumers (66.6%) washed the chicken meat under running tap water and eight (44.4%) used cutting boards. Also, only five consumers washed their hands properly prior to or during meal preparation. Cross-contamination events were detected in four kitchens, between the raw chicken and two cutting boards, two sinks and one kitchen cloth. The poultry samples presented different levels of contamination (< 4.0 × 101 CFU/g to 2.2 × 103 CFU/g), being some poultry with lower Campylobacter loads the origin of three cross-contamination events during food preparation. Both C. jejuni and C. coli were recovered. Molecular typing by PFGE showed a high diversity among the isolates. There were different explanations for the practice of cleaning and rinsing chicken, but, in general, it is an habit linked to what they have learned from their families. These results highlight the potential for the dissemination of Campylobacter strains in the domestic environment through the preparation of chicken meat and the need to raise awareness among consumers for an appropriate handling of raw poultry in order to decrease the risk of campylobacteriosis.

First Report of Botryosphaeria dothidea Causing Stem Canker and Dieback of Gleditsia sinensis in China.

Gleditsia Sinensis Lam is a kind of legume perennial woody plant, which is a traditional Chinese medicine with high economic and ecological value. Its planting area is about 0.1 million ha. InJuly 2018, symptoms of stem blight were observed on G. sinensis in An shun (26.072311°N, 106.097433°E), Guizhou province (southwestern China). Symptoms included stem canker and dieback, twig blight and extensive vascular discoloration, with incidence rate of 45 to 70%. Samples from plants with symptoms were washed with running tap water, surface sterilized with 2% sodium hypochlorite and then 75% ethanol, rinsed in sterile distilled water, plated on potato dextrose agar (PDA) and incubated at 28°C. Fungal isolates developed copious white aerial mycelium that became dark grey after four to five days, and formed black pycnidia after 23 days. Single hyphal tip cultures of putative 27 isolates were stored in the culture collection (CMW) of the Urban Modern Agriculture Engineering Research Center at the Kunming University. Conidia forming on pycnidia were one-celled, hyaline, aseptate, and fusiform, with dimensions of 20.3-25.9 μm x 4.2-6.5 μm (average 22.5 x 5.5 μm) (sixty conidia were measured). DNA sequence analysis of the ribosomal internal transcribed spacer regions ITS1-5.8S-ITS4, β-tubulin, and the translation elongation factor 1-alpha (EF1-α) were performed. BLAST searches at GenBank showed the highest nucleotide sequence identity with Botryosphaeria dothidea reference sequence (ITS: >99.82%, KR708996; EF1-α: 99.62%, KP183214; β-tubulin: > 99.67%, KU306116). Representative sequences of isolates from these regions were deposited in GenBank (ITS: Accession No. MT449017; EF1-α: Accession No. MT454342; β-tubulin: Accession No. MT454343). Morphological and molecular results confirmed this species as B. dothidea (Aguirre et al. 2018). Pathogenicity tests were conducted by stem inoculation of two-year-old G. sinensis seedlings. Mycelial plugs (2-3 mm in diameter) from actively growing colonies of B. dothidea (PDA) were applied to same-size bark wounds on the middle point of the stems. Inoculated wounds were wrapped with Parafilm. Control seedlings received sterile PDA plugs. Inoculated and control seedlings (9 each), and kept in the greenhouse (28℃, 80% humidity); After 10 days, all of the inoculated plants showed dark vascular stem tissue, and the controls remained healthy. After 30 days, all of the inoculated but none of the control G. sinensis seedlings showed leaf wilting and tissue discoloration. B. dothidea was re-isolated from symptomatic tissues, thus fulfilling Koch's postulates. No symptoms were visible in the control seedlings, and no B. dothidea was re-isolated from the control seedlings tissues. B. dothideais a member of Botryosphaeriaceae, commonly associated with cankers and dieback of woody plants. B. dothideahas been reported as a pathogen causing stem dieback and branch canker on Malosma laurina (Aguirre et al. 2018), Helwingia chinensis (Yu et al. 2012), and blueberry (Choi 2011; Yu et al. 2012). To our knowledge, this is the first report of B. dothidea on G. sinensis in China.

Awareness and Practices of Alginate Disinfection Protocol Among Dental Practitioners of Karachi City

Objective: To determine the awareness and practices about disinfection of alginate impression among dental practitioners of Karachi city. Study Design and Setting: This cross-sectional study was conducted at a private sector teaching hospital and private clinics, both located in Hamdard University Dental Hospital, Karachi from the period of April, 2017 to April, 2019. Methodology: A Questionnaire was designed by subject specialist in light of Australian Dental Association guidelines for cross infection prevention and disinfection for dental offices and laboratory. The questionnaire comprised of demographic details including age, gender, years of clinical experience; awareness of disinfection protocols such as self-protection precautionary measures while pouring the alginate impressions; practices of disinfection in the Laboratory; Practice of personal protection and details regarding the attendance of CDE (Continuing Dental Educations) seminars. The data was analyzed on Statistical Package for Social Sciences SPSS version 20. Results: Among total subjects of 186, 78(41.9%) male and 108 (58.06%) female participants were included in this study. In present study the awareness of disinfection of alginate was observed in 171(91.8%) subjects while 15(8.2%) were not up to the mark. Regarding precautionary measures n=142 (83.0%) participants used gloves,. Considering the preferred method and duration of disinfection 56.4% used running tap water for gross cleaning of impression for 5 minutes while 43.6% marked spraying with 2% sodium hypochlorite as preferred disinfection method. Conclusion: majority of the participants of this study were aware regarding the ADA guidelines of disinfecting the alginate impressions while there was a dearth of practice observed for disinfecting the alginate impressions and were not used the prescribed disinfectant to disinfect the dental impressions

Primary management of burn injuries: Balancing best practice with pragmatism.

Management of burns is an often-neglected area in training from undergraduate to specialist level. There is, however, a high burden of injury that affects a largely vulnerable population, for example, children, the elderly and epileptics. This CPD article highlights that first aid should include cooling the burn with cool running tap water up to 3-hours post injury (Burnshield may be used if cool running water is not available); removal of all blisters facilitates accurate assessment of the burn size and depth; formulas exist for the resuscitation of acute burn injuries of more than 10% – 15% total body surface area and prophylactic antibiotics should not be administered to patients with acute burns as the prevention of infection should lie with good wound care (including good wound cleaning and the use of topical antimicrobial dressings). A standardised approach to pain management with an incremental pharmacological approach should be followed whilst considering other issues such as neuropathic pain, anxiety and depression.

On the possibility of obtaining high-quality lyophilized collagen hydrolysate and its applicability in the sausage production

Abstract The paper discusses the issues of rational use of secondary collagen-containing raw material in the food industry, namely cattle by-products. The technology of obtaining collagen hydrolysate from this type of raw material with the subsequent freeze-drying is proposed, which maximizes the level of preservation of native properties and the expands the scope of its application. The paper presents the stages of obtaining lyophilized collagen hydrolysate by biomodification of frozen cattle lips and ears. The preparation of lyophilized collagen hydrolysate was carried out as follows: the lips and ears were preliminary washed in running tap water for 20 minutes in order to remove mucus and impurities, muscle and adipose tissue was also removed; after draining (15-20 minutes), the raw material was ground on a top with a grid diameter of 2-3 mm. At the next stage, the crushed lips and ears were treated with a solution of Protepsin enzyme preparation in the ratio of the raw material to solution 1:2 and kept for 0.5-1.5 hours at a concentration of the enzyme preparation of 0.06% by weight of the raw material, while stirring periodically (at 17-22 °C). The filtrate from the product was separated on a sieve. Processed as described above, the raw material was subjected to freeze-drying. Evaluation of the temperature and duration of freeze-drying of collagen hydrolysate was carried out at a drying layer thickness of 5.0 mm and different drying temperatures: 20 °C, 30 °C, 40 °C, and 50 °C.

Phytophthora cactorum as a Pathogen Associated with Root Rot on Alfalfa (Medicago sativa) in China.

Alfalfa (Medicago sativa) is the largest grown pasture crop in China due to its economic and ecological importance. During the summer season from June to August in 2018, stunted plants was frequently observed in alfalfa fields that have been established for two years in Jinchang, Gansu Province. The disease incidence of root rot ranged from 40% to 50%. Taproots of stunted plants showed red-brown to dark brown discolorations, and lateral roots were poorly developed. Shoots wilted with rotted taproots and lateral roots in severely affected plants. Twenty symptomatic plants were collected and transported to the laboratory for pathogen isolations. Roots were washed under running tap water, cut into 2 to 3 mm pieces (40 pieces each plant), and then sterilized in 75% ethanol for 2 mins followed by three times washing with autoclaved distilled water. Surface dried pieces on autoclaved filter papers were put onto water agar and also a Phytophthora selective medium P5ARP(H) (Jeffers and Martin 1986). The plates were incubated at 22°C for 3 to 5 days and then the growing hypha were subcultured onto potato dextrose agar (PDA). Thirty-two Phytophthora-like isolates were obtained and showed similar morphologies on PDA. Five isolates picked randomly were purified by single-hyphal-tip and plugs (4 to 5 mm) from PDA cultures were incubated in petri dishes with autoclaved distilled water at 22°C for 5 days. Sporangia, chlamydospores and oospores were examined. Sporangia were usually ovoid and sometimes appeared ellipsoid, with the length of 30.5-39.1 μm and width of 23.4-27.8 μm. The diameter of chlamydospores was 29.6 to 42.5μm. Oospores had a diameter of 23.6 to 30.2 μm. The isolates were tentatively identified as P. cactorum based on these morphology characteristics (Montealegre et al. 2016). DNA of these isolates were extracted and PCR amplifications of the rDNA ITS region and cytochrome oxidase subunit I (Cox I) (Kroon et al. 2004) were conducted. Sequences of these isolates were then compared with reference sequences in GenBank using BLAST search. The 866-bp ITS sequences had a sequence identity of 99% to 100% with P. cactorum (e.g. accession nos. EU662221, KJ128036). In addition, the 663-bp CoxI sequences showed 100% sequence identity with three P. cactorum isolates (accession nos. AB688156, HQ708234, EU660851). The ITS and CoxI sequences of one representative isolate Phy.c2 have been deposited in GenBank with the accession no. MT280033 and MT344138, respectively. Pathogenicity of the five isolates (Phy.c1-Phy.c5) were determined on two-week-old alfalfa seedlings (cv. Longdong) grown from seeds. Inoculums were prepared by subculturing agar plugs from edges of PDA cultures into the flask with autoclaved millet seeds, and incubated at 22°C in darkness for two weeks and shaken by hand every two days to ensure uniform colonization. Seedlings were transplanted into pots (12 cm x 12 cm) filled with autoclaved potting mix infested with millet-seed inoculum of each isolate at a rate of 0.5% (w/w). Control seedlings for comparison were transplanted into pots with uninfested potting mix. There were five seedlings per pot and twelve replicate pots for both inoculated and noninoculated treatments, and pots were kept under controlled environment room (22°C, 12 h photoperiod and 65% relative humidity) that were watered every two days to free draining. 87%~92% of the inoculated plants showed stunted symptoms with poorly developed and brown-discoloured roots three weeks after inoculation while the control plants were healthy with no root disease symptoms. To fulfil Koch's postulates, re-isolated cultures from discoloured root tissues were confirmed as the inoculated isolates by morphological examination and ITS sequencing. The five-purified isolates were submitted to the Grassland Culture Collection Center, Lanzhou University, with the accession nos. LZU-MsR-Phy.c1-Phy.c5. To our knowledge, this is the first report of P. cactorum as a pathogen of root rot on alfalfa in China. Phytophthora spp. has been reported causing root rot on alfalfa in America, Australia and Canada, and other legumes such as chickpea, and many other crops worldwide (Musial et al. 2005; Tan and Tan 1986; Vandemark and Barker 2003), and P. cactorum was reported as a root rot pathogen on lavender in China (Chen et al. 2017). P. cactorum may be a significant pathogen associated with root rot in major commercial alfalfa-producing areas in China where are based on flood-irrigation during the growth season.

Current Trends in Treating Acanthamoeba Keratitis: A Brief Narrative Review

The Author, after examining the historical evolution of scientific knowledge and treatment of severe Acanthamoebic keratitis, presents this brief review on the treatment of this serious eye disease, relatively frequent in patients with corneal contact lenses. Therapy of Acanthamoeba keratitis is always very long and demanding. Its management requires adequate experience because it is not always easy to evaluate the response to treatment and complications can be very serious and difficult to manage. Resistance to therapy can also occur during treatment and must be distinguished from drug-induced toxicity. In cases where no improvement is obtained with maximum medical therapy, it is advisable to repeat the corneal sampling and proceed to new laboratory tests for Acanthamoeba, bacteria and fungi. Prevention, which always remains of fundamental importance, is practically based on avoiding contact of the corneal lens with contaminated water, since this Acanthamoeba has a ubiquitous diffusion. It is therefore recommended to always avoid the use of corneal contact lenses in the pool or in the shower, not to wash them under running tap water and to frequently replace the relative container of these lenses.&#x0D; Key Words: Acanthamoeba Keratitis, Corneal ulcer, Contact lenses.