Brown Discoloration Research Articles

Identification of romaine lettuce (Lactuca sativa var. longifolia) Cultivars with reduced browning discoloration for fresh-cut processing

Discoloration (browning) represents a major challenge that limits the quality and shelf life of fresh-cut lettuce. In this study, we aimed to find romaine lettuce (Lactuca sativa var. longifolia) accessions with low browning potential. Midribs of 14 accessions (11 cultivars, two breeding lines, and a single plant introduction) were shredded and packaged in perforated bags for five days. Images of processed samples were captured daily and analyzed with computer vision technology to quantify browning intensity via L*a*b* color values and browning index (BI). Enzymatic activity[phenylalanine lyase (PAL), peroxidase (POD), and polyphenol oxidase (PPO)] and total phenolic content (TPC) were measured daily. After five days in storage, the accessions in the Tall Guzmaine and Parris Island Cos pedigree groups exhibited the greatest and least browning, respectively. In addition, while the PAL, POD, and TPC increased substantially over time, the PPO of twelve accessions fluctuated with only minor increases. For all accessions, the temporal increase of PAL, POD, and TPC showed significant, positive correlation to browning progression. Comparing between accessions, those that had greater amounts of accumulated PAL and smaller amounts of POD tended to have a greater amount of browning after five days of storage, despite relatively low correlation coefficients. However, the accumulation of TPC and PPO was not correlated to browning severity after five days of storage. This systematic study provides lettuce growers and breeders with guidance for selecting accessions with limited browning, and it supplies researchers in plant physiology and genetics with more information on the roles of enzymes in the lettuce browning process.

First Report of Charcoal Rot Caused by Macrophomina phaseolina on Faba Bean in China

Faba bean (Vicia faba L.) is an important legume crop in China. Soil-borne charcoal rot, caused by Macrophomina phaseolina (Tassi) Goid, is a devastating disease of many crops, including legumes. In late March 2018, wilt symptoms similar to those of charcoal rot were observed in faba bean plants (‘Yundouzao 6’) in a field located in Jianshui County (23.38° N, 102.49° E), Yunnan Province, China. This disease caused serious yield loss of faba bean. Infected plants initially showed leaf chlorosis and wilting, and the plants eventually died with the leaves remaining attached. When affected plants were examined, the basal stem pith was rotted, and a brown discoloration was observed after splitting the roots and stems. Symptomatic plants were sampled to isolate the pathogen. Three fungal isolates were obtained (FMP8-1, FMP8-2, and FMP8-4) and displayed rapid mycelial growth on potato dextrose agar at 28°C with a 12-h light regime. The mycelia were initially hyaline and turned from gray to dark with age. Masses of dark oblong microsclerotia (46 to 96 × 61 to 129 µm) formed on the cultures 3 days after incubation. On the basis of these morphological characteristics, the three isolates were presumptively identified as M. phaseolina (Smith and Wyllie 1999). To confirm this identification, the three isolates were molecularly analyzed using primers specific for M. phaseolina: MpKF1/MpKR1, which yielded a 350-bp target fragment (Babu et al. 2007). Additionally, the internal transcribed spacer (ITS) regions of the three isolates yielded ∼600-bp PCR fragments using the universal primers ITS4/ITS5 (White et al. 1990), and the resulting sequences (accession nos. MH323404, MH323405, and MH323406) showed 99 to 100% identity with several known M. phaseolina strains using a BLAST algorithm-based analysis, including strains CIAT519 (KR012878) and CPC13084 (KF95698) isolated from Phaseolus vulgaris; CPC21464 (KF951750) and CPC21386 (KF951701) from Arachis hypogaea; CPC21514 (KF951775), CPC21517 (KF951778), and CPC21450 (KF951739) from Vigna unguiculatain; CBS231.33 (KF951631) from Saccharum officinarum; CJMR135 (MF480342) from Corchorus olitorius; and CPC13086 (KF951700) from Solanum melongena. Thus, this fungus has a broad host range. Both morphological and molecular characteristics confirmed that the three isolates were M. phaseolina. Pathogenicity tests were performed on 2-week-old faba bean seedlings using three inoculation methods (detached leaf, cut stem, and stem puncture) and conducted in three replications with five seedlings per replication (Keeling 1982). All inoculated leaves, stems, and plants showed necrotic symptoms 4 days after inoculation. No symptoms were observed on the control leaves, stems, or plants. M. phaseolina was reisolated from all inoculated leaves, stems, and plants, fulfilling Koch’s postulates. The three isolates were also pathogenic on other legume crops, including soybean (Glycine max), mungbean (Vigna radiata), pea (Pisum sativum), and grass pea (Lathyrus sativus). Although M. phaseolina has been reported on legume crops including soybean, mungbean, and adzuki bean in China, to our knowledge this is the first report of M. phaseolina causing charcoal rot on faba bean in China.

PB2048 ACUTE IRON OVERLOAD WITH IRON CARBOXYMALTOSE:CASE REPORT

Background:Iron deficiency (ID), with or without anemia, represents a major global health problem afecting more than 2 billions people worldwide. Oral iron treatment always becomes the first choise for the treatment of iron deficiency. An other treatment option is parenteral iron therapy and generally is prefered for patients with oral iron treatment is not effective. But today there is a number of conditions where intravenous iron use is well‐established or increasingly considered as first‐option approach. All these developments in the parenteral iron area show strong probability of increasing use of parenteral iron drugs for anemic patients.Aims:We this case report we aim to share our clinical case in a rare situtionMethods:Case reportResults:Our patient is 36 years old female and she was diagnosed as iron deficiency anemia in an outpatient clinic. She was treated with ferric carboxymaltose. One month later she was reevaluated her for treatment response and her anemia was treated succesfully but her serum ferritin value was measured as 3500 ng/ml. It was found unusual and she was referred to our clinic for evaluation of hiperferritinemia. We evaluated the patient and no pathological findings were found other than skin brown discoloration. We measured patient's serum ferritin level again and ferritin level was high (3000 ng / ml) as one month before. When we investigated ferric carboxymaltose dose, we saw that in contrast to the ferric carboxymaltose recommended maximum 2000 mg dose for adult patients, it was administered to the patient at a total dose of 8000 mg in two months period as weekly 1000 mg dose.The patient underwent cardiac T2 ∗ magnetic resonance imaging (MRI), liver T2 ∗ MRI, brain MRI. The patient MRI showed significant increase iron in the liver and spleen, whereas increased iron accumulation in brain and heart was not detected with MRI.Deferasirox was started at a dose of 20 mg / kg / day. The ferritin level decreased to 1600 ng / mL from 3000 mg/mL level at the second treatment week with iron chelation therapy. Patient's skin color was not changed significantly. We decide to continue until the serum ferritin value decreases below 1000 ng/mL.6 months after deferasirox treatment ferritin levels go down step by step and reached to 700 ng/L level. We repeated liver T2 ∗ MRI and there is significant reduction in liver iron accumulation. And her skin discoloration was decreased.Summary/Conclusion:There is no literature data about overdose of ferric carboxymaltose case. We found only two postmarketing adverse case report of national pharmaceutical agencies. First case is a hypophosphatemic osteomalacia case resulted from 8000 mg ferric carboxymaltose usage in 4 month period. The second case is from in France. Ferric carboxymaltose,18000 mg total cumulative dose in six months is received and presented as movement disorder complication. It was reported that iron chelation therapy partially alleviates gait disturbance and relieves it from the dependency on the bed which emerged in the first period in this case. According to these data, first step in ferric carboxymaltose overdose cases;it should be aimed to prevent the development of neurological complications and bone complications. The best way of to prevent these complications can be done with rapid beginning iron chelation therapy. Our case is the first reported case in our country and third case report in the world.

Occurrence of Verticillium Wilt Caused by Verticillium dahliae on Okra (Abelmoschus esculentus) in North China

Okra (Abelmoschus esculentus L. Moench) belongs to the Malvaceae family, which is a fairly new vegetable crop currently grown in a large area in China from Hainan to Hebei province. During 2015 to 2017, wilted okra plants were observed from May to September in an okra seed breeding base located in Beijing’s Fangshan District. The affected plants showed typical wilt symptoms, including gradual leaf yellowing, wilting, and dark brown discoloration of the vascular tissue. The affected plants later died as the disease progressed. Field surveys were conducted to investigate the disease in okra in north China from early June to late July 2018. The disease was found sporadically but had spread to every okra-growing planting area in north China. To identify the pathogen associated with this disease, basal stems of diseased plants were washed under tap water, and then the discolored vascular tissue was cut into fragments of 0.5 cm in diameter, which were surface disinfested with 3% sodium hypochlorite for 10 min, rinsed thoroughly with sterile distilled water, dried on aseptic filter paper, and then placed onto potato dextrose agar medium. After incubating the plates in the dark for 5 days at 26°C, typical Verticillium dahliae colonies were observed, characterized by development of white mycelium that eventually turned to black owing to formation of microsclerotia. Six isolates were initially identified on the basis of verticillate conidiophore morphology and production of microsclerotia. Furthermore, the internal transcribed spacer region of rDNA of these isolates were amplified using ITS1/ITS4 primers, described by White et al. (1990), and the amplicons were the same in sequence (GenBank accession no. MG910491). BLAST analysis of the 541-bp amplicon showed 100% identity with the reported V. dahliae strains (GenBank accession nos. GU461618.1 and FJ475122). Pathogenicity tests of the six isolates were performed on 20-day-old seedlings of okra K1, K6, K11, and K20 accessions (30 seedlings) using a stem-injection inoculation method with a spore suspension of 1.0 × 10⁶ conidia/ml (Bolek et al. 2005). The inoculated seedlings were kept in a greenhouse at 25°C. Seedlings treated with sterile distilled water served as a control. At 10 days postinoculation, all inoculated plants exhibited wilt symptoms with leaf scorching, wilting, and abscission. No symptoms were observed on control plants. The pathogen was reisolated from all inoculated plants, thus fulfilling Koch’s postulates. To our knowledge, this is the first report of Verticillium wilt on okra in China. Considering the threat of okra Verticillium wilt, a project should be conducted for screening and breeding resistant varieties to manage the disease.

A comprehensive analysis of the Lactuca sativa, L. transcriptome during different stages of the compatible interaction with Rhizoctonia solani

The leafy green vegetable Lactuca sativa, L. is susceptible to the soil-born fungus Rhizoctonia solani AG1-IB. In a previous study, we reported on the transcriptional response of R. solani AG1-IB (isolate 7/3/14) during the interspecies interaction with L. sativa cv. Tizian by means of RNA sequencing. Here we present the L. sativa transcriptome and metabolome from the same experimental approach. Three distinct interaction zones were sampled and compared to a blank (non-inoculated) sample: symptomless zone 1, zone 2 showing light brown discoloration, and a dark brown zone 3 characterized by necrotic lesions. Throughout the interaction, we observed a massive reprogramming of the L. sativa transcriptome, with 9231 unique genes matching the threshold criteria for differential expression. The lettuce transcriptome of the light brown zone 2 presents the most dissimilar profile compared to the uninoculated zone 4, marking the main stage of interaction. Transcripts putatively encoding several essential proteins that are involved in maintaining jasmonic acid and auxin homeostasis were found to be negatively regulated. These and other indicator transcripts mark a potentially inadequate defence response, leading to a compatible interaction. KEGG pathway mapping and GC-MS metabolome data revealed large changes in amino acid, lignin and hemicellulose related pathways and related metabolites.

Systemic lipofuscinosis associated with a lesion of autophagic vacuolar myopathy in the diaphragmatic muscle of a cow

A 7-year-old Holstein-Friesian dairy cow that was culled because of traumatic motor dysfunction due to accidental falling down showed a variety of gross postmortem findings, such as fracture of the caudal vertebra, massive hemorrhage in the pelvic cavity, edematous swelling of the right sciatic nerve, and brown discoloration of the masseter muscle, tongue, cardiac muscle, diaphragmatic muscle, liver, adrenal cortex, and intestinal wall. Histopathologically, the discolored and some other internal organs/tissues exhibited varying degrees of intracellular deposition of lipofuscin granules. A small number of eosinophilic proteinaceous granules, along with lipofuscin granules, were found in the smooth muscle fibers of the intestinal muscular layer. The diaphragmatic muscle showed a lesion of autophagic vacuolar myopathy, representing frequent occurrence of autophagic vacuoles that contained amorphous materials, lipofuscin granules, and eosinophilic proteinaceous droplets. It is presumable that the pathogenetic mechanism of this systemic lipofuscinosis had a close relationship with ischemia due to anemic condition caused by pelvic hemorrhage. This bovine case was considered a newly identified variant of systemic lipofuscinosis associated with an autophagic vacuolar lesion in the diaphragmatic muscle.

Ochronotic arthritis: one case report and literature review

Objective To summarize the clinical manifestation, diagnosis and treatment methods of ochronotic joint disease, thus to improve grassroots medical workers awareness of ochronotic arthritis. Methods The clinical data of 1 case of ochronotic arthritis in the Second Municipal Hospital of Weihai Affiliated to Qingdao University and literature data were analyzed. Results The male patient, 65 years old, because the right femoral neck fracture admitted to accidentally falls in patients with bilateral antihelix ear cartilage and local black pigmentation, resection of the femoral head cartilage surface, black, dark color round ligament, partial hip joint capsule was dark, multi segmental lumbar vertebral intervertebral space narrowing disc hardening, laboratory urine silver nitrate and ammonium hydroxide examination appear dark brown discoloration, confirmed the diagnosis of patients with ochronotic joint disease. Conclusion We should improve cognition of ochronotic joint disease, examination by medical history, clinical manifestations, laboratory and imaging, diagnosis of ochronotic arthritis, thus to reduce the rate of misdiagnosis. Key words: Joint diseases; Ochronosis; Case reports; Review literature as topic

Fusarium solani causing stem rot and wilt of lucky Bamboo (Dracaena sanderiana) in India-first record

Lucky Bamboo (Dracaena sanderiana Sander ex Mast) is a tropical evergreen perennial woody, shrubby species. This is native of Cameroon area in tropical West Africa. This has flexible strap-shaped leaves and slender stems. Being an indoor house plant it grows very well under indirect lighting and have gained popularity because of adoption of new age culture. It multiplies readily by stem cuttings. In year 2014–2017, stem rot and wilt symptoms were observed in D. sanderiana cuttings. The first observed symptoms were yellowing in leaves with wilting. In due course of time the leaves turned dry and light brown with necrosis. On splitting open, rotted areas were seen in the middle with a visible brown discoloration in cortical region. The infected plants died within a few weeks. The fungal pathogen Fusarium was observed and isolated from the collar, roots, stems. This was morphologically confirmed as Fusarium solani and also through amplification and sequencing in ITS region, which showed cent percent similarity with the sequences of F. solani available in NCBI genbank. As per record in literature this is the first record of stem rot and wilt caused by F. solani in plants of D. sanderiana from India.

First Record of Erysiphe elevata on Plumeria rubra in Taiwan

Plumeria rubra L., native to tropical Central America, is a widely cultivated ornamental woody plant. In two municipalities in northern Taiwan we observed brown discolorations irregularly following veins on the abaxial surface of green or yellowing lower (old) leaves of two outdoor plants of P. rubra. The upper side did not show symptoms or diffuse yellow spots. About 80% of leaves exhibited superficial mycelia and conidiophores typical of a powdery mildew. Specimens collected from a garden plant in Taipei City on 31 May 2018, and from a pot plant in Taoyuan City, Zhongli District, on 7 June 2018, were deposited under the collection numbers R. Kirschner 4647 and 4651 in the National Museum of Natural Science, Taichung, Taiwan (TNM). Fresh specimens were mounted in approximately 10% KOH and observed with light microscopy. The mycelium was hypophyllous, forming irregular patches. Hyphae were hyaline, walls thin, smooth, 3 to 8 μm wide, with hyphal appressoria single or opposite in pairs, nipple shaped or lobed. Conidiophores were 35 to 76 μm long, 7 to 9 μm wide, erect, straight or curved at the base, mostly three-celled, occasionally one- or two-celled. Foot cells were rough at the base, 24 to 35 μm long, 6 to 8 μm wide, with basal septum mostly even with the surface of the supporting hypha. Conidia were solitary, ellipsoidal to cylindrical, smooth, 28 to 35 μm long, 13 to 16 μm wide, and germination was not observed. The specimens were compared with a specimen (R. Kirschner 1945, TNM) of E. elevata on Catalpa sp. (Bignoniaceae), collected in Germany, Frankfurt am Main, Bergen-Enkheim, 18 July 2004. The anamorphic specimens from Catalpa sp. in Germany and P. rubra from Taiwan were morphologically indistinguishable. The internal transcribed spacer (ITS) region of rDNA of both Taiwanese specimens was amplified with the primers ITS1F/ITS4 and sequenced. The 660-bp-long ITS sequences of R. Kirschner et al. 4647 (GenBank MH507182) and 4651 (GenBank MH507183) were 100% identical to each other and 99 to 100% identical to the 10 available sequences of E. elevata in GenBank (Meeboon and Takamatsu 2017), whereas the similarity was 98% or lower for other species. Erysiphe sp. (labeled as E. elevata in GenBank under MF092833) recorded on Plumeria pudica by Suarez et al. (2017) had a similarity of 97% (582/597 bp). The sequence MF092833 contained seven unclear positions (“N”), indicating technically insufficient quality. Because of the high similarity of sequences and anamorph morphology, the specimens on P. rubra from Taiwan are conspecific with E. elevata. It seems to be limited to the lower side of the older leaves, where it forms inconspicuous mycelia so that the brown scribble-like discolorations along the leaf veins are more diagnostic symptoms than the mycelial colonies. Powdery mildew of Plumeria leaves is often reported anecdotally, but confirmed reports are lacking. Recently, Suarez et al. (2017) identified the causative agent as the genus Erysiphe on P. pudica in the United States. Because P. rubra was introduced to Taiwan in the 17th century and the first hybrid in 1967 (Hsueh and Yang 2009), the pathogen may have gone unnoticed for a long time. According to our observation, however, the powdery mildew appears to be less frequent than the recently reported rust Coleosporium plumeriae (Chung et al. 2006) and might have been recently introduced. The fungus may be involved in an accelerated leaf-fall, particularly in connection with additional stress factors such as drought or coinfection with the rust fungus.

Successful Management of Nitrite Poisoning in Crossbred Dairy Calves

Nitrate/nitrite poisoning in dairy cattle occurs when there is an abrupt change in diet from the introduction of a bulk feed source with high nitrate levels. Over fertilization of fodder with untreated manure and occurrence of drought season followed by a rainy season could result in higher level of accumulation of nitrate in plants. A sudden onset of illness was reported in eight crossbred dairy calves (between 8–11 months of age) at University Livestock Farm and one among the affected calf died after showing symptoms of discomfort, excessive salivation, tympany, dyspnoea, sternal recumbency progressing to lateral recumbency, pedalling movements and, stiffness of the limbs and neck. On clinical examination, the affected animals exhibited sialorrhoea, tympany, increased pulse and respiratory rates, laboured breathing, brownish discoloration of mucous membrane and loss of co-ordination. Microscopical examination of rumen fluid revealed normal protozoan motility. Blood colour was dark chocolate brown in all the affected animals and coagulation time was delayed. Animals had a history of ad libitum feeding with chopped lush green fodder grasses namely hybrid Napier and fodder maize that were harvested from cattle slurry irrigated fodder plots. Based on the history and clinical signs noticed, the case was presumptively diagnosed as nitrate/nitrite poisoning, later confirmed with diphenylamine test. All the animals were treated with intravenous administration of 1 per cent methylene blue @ 1–2 mg/kg BW. Subsequently the dairy calves showed an uneventful recovery from the condition.

The major safeguard for fluorosis using age-appropriate toothpaste

Until now it is well established that fluoride is an important ingredient to combat dental caries, working both systemically and topically. As the action of fluoride is dose-related and it has a cumulative action, it’s over dosage can cause serious toxic effect. Fluorosis is one of the chronic toxic effects of fluoride that appears as a developmental disorder of dental enamel. Fluorosis develops due to multiple and continuous exposures to high concentration of fluoride in low dosage. Therefore, the development of fluorosis is not only dependent on the dose but also greatly reliant on length and timing of fluoride exposure. This paper presented a case of dental fluorosis of a 9 years old girl who had a history of swallowing of toothpaste during tooth brushing when she was younger than 3 years. Clinically fluorosis appears as subsurface hypo mineralization or porosity (mottling enamel) of teeth where usually fine white to chalky opaque to brown discoloration of enamel occur which was typically seen in all of my patient’s permanent teeth (all permanent incisors and all permanent first molars) present at that time in her mouth while all of her deciduous teeth (all deciduous canines and molars) present at that time in her mouth were absolutely sound. Micro-abrasion is one of the procedures to treat the fluorosis teeth. In this case micro-abrasion was performed successfully.

Genotypic Variation in Apple Rootstock Low Temperature Tolerance During Spring and Fall

To identify genotypes of apple (Malus ×domestica) rootstock with vulnerability to low temperature, we measured the low temperature tolerance of xylem, phloem and cambium in 2-year-old shoot pieces from cultivars Budagovsky 9 (B.9), M.7 EMLA (M.7), M.9 EMLA (M.9), Geneva® 41 (G.41), Geneva 30 (G.30), Geneva 214 (G.214), Geneva 814 (G.814), and Geneva 935 (G.935), as well as six advanced selections in the Geneva (G.) series and three in the Vineland (V.) series. From Oct. 2013 to Apr. 2014, injury was measured as a 0–10 rating based on percentage of discolored cross-sectional xylem and phloem, and cambial length and circumference with brown discoloration, with 0 indicating no browning and 10 indicating browning in the entire tissue. From Oct. 2014 to Apr. 2015, injury was measured as xylem, phloem and cambium browning using a similar rating scale that accounted for both the percentage of browned tissues and the intensity of browning. Following exposure to −35 to −40 °C, many genotypes, including ‘M.7’, ‘M.9’, ‘G.935’, G.4011, G.4292, G.5087, and V.5, had only partial xylem injury in the fall, whereas others, ‘M.7’, ‘G.41’, ‘G.214’, and G.4011, had more extensive xylem browning at −30 °C and colder. ‘G.30’ had moderate to severe xylem browning at −15 to −19 °C. In late October of both years, G.4013 exhibited severe phloem browning at relatively high temperatures, but accrued additional hardiness by Nov. 2014, whereas genotypes ‘B.9’, ‘M.9’, ‘G.30’, and ‘G.41’ developed considerable phloem hardiness by late October with no additional increase in hardiness in November. Geneva and Vineland genotypes exhibited a low degree of susceptibility to injury at −35 to −40 °C in Jan. 2014 and Mar. 2015. Shoot hardiness in Apr. 2014 and 2015 was highly variable between the 2 years, with severe browning of xylem and cambium at −40 °C in every genotype sampled in Apr. 2014, but not in Apr. 2015. ‘M.9’ and G.3902 appeared to be the least vulnerable to injury in April, whereas ‘G.30’, ‘G.41’, ‘G.814’, G.4292, and G.5257 seem more likely to suffer injury in spring. ‘G.30’ had tender xylem in both fall and spring, G.4013 had the least hardy cambium and phloem in fall, and G.5257 the least hardy cambium in the spring. These genotypes are vulnerable to damaging temperatures during fall acclimation and spring deacclimation.

Histochemical observations and gene expression changes related to internal browning in tuberous roots of sweet potato (Ipomea batatas)

The mechanism underlying internal browning (IB), or brown discoloration, of the central region of tuberous roots of sweet potato (Ipomoea batatas) was examined. IB disorder begins in roots from approx. 90 days after transplanting, and the severity increases significantly with time. IB damage initially occurs in cells around the secondary vascular tissue, and the area per cell occupied by starch grains in this region was larger than in the unaffected region. High levels of reducing sugars, polyphenol oxidase (PPO) activities, chlorogenic acid, and hydrogen peroxide (H2O2) were detected in cells from the IB damaged regions. The content of sugar and polyphenols was higher in disks (transverse sections) with larger amounts of damaged tissues than in disks of sound root. The transcript levels of acid invertase (IbAIV) tended to be higher with greater IB severity, whereas fluctuation patterns of ADP-glucose pyrophosphorylase (IbAGPase), granule bound starch synthase (IbGBSS), and starch branching enzyme 1 (IbSBE1) were lower with higher IB severity. These observations suggest that the incidence of IB disorder in sweet potato is largely dependent on the excessive generation of reactive oxygen species (ROS) in cells around the secondary vascular tissues due to the abundant accumulation of sugar and/or starch grains during the root maturation period.

First report of Agaricus sect. Brunneopicti from Pakistan with descriptions of two new species

Agaricus is a genus of saprophytic fungi in the Basidiomycota. As part of ongoing studies on the genus Agaricus in Pakistan, we here describe and discuss two new species, A. pakistanicus sp. nov. and A. sparsisquamosus sp. nov. Morphological characters and phylogenetic analysis of ITS sequences were used to clarify their taxonomic affinities. Phylogenetic analysis indicated that both species belong to A. sect. Brunneopicti. This palaeotropical section, which has been recently reconstructed, is reported in Pakistan for the first time. Interestingly, these two new species exhibit a reddish or brown discoloration. With these two new species, the distribution of this section, which was previously restricted to tropical and subtropical humid regions, now extends to semi-arid and desert climate regions and the total number of species in the section increases to 20, including eight unnamed species.

Outbreak of Bovine Herpetic Meningoencephalomyelitis in Southern Brazil

Background: Herpetic meningoencephalitis is an infectious contagious disease worldwide distributed, most often caused by bovine alphaherpesvirus type 5 (BoHV-5), although bovine alphaherpesvirus type 1 (BoHV-1) may occasionally be the causative agent. The disease is characterized by subacute to acute clinical onset, often affecting animals submitted to stressful situations. Clinical signs are mainly neurologic due to meningoencephalitis and cortical necrosis. The involvement of the spinal cord has also been reported, however in BoHV-1 associated disease only. The aim of this report is to describe an outbreak of bovine meningoencephalomyelitis associated to BoHV-5.Case: In August 2017, nine 1-year-old calves died in a beef cattle farm with a flock of approximately 400 bovines. The animals presented neurological clinical signs characterized by excessive salivation, nasal and ocular discharges, incoordination, apathy, head tremors, head pressing, wide-based stance, recumbency followed by convulsions and paddling. According to the owner and referring veterinarian, affected animals displayed severe clinical signs with rapid progression and often leading to death in up to seven days. Four of these calves were submitted for necropsy, and gross lesions were present in the brain, characterized by mild to moderate multifocal hemorrhagic and soft areas. On cut surface, extensive areas of dark brown discoloration and malacia were observed. Histologically, lesions were characterized by extensive areas of liquefactive necrosis in the cerebral cortex grey matter, associated with inflammatory infiltrates composed of neutrophils, lymphocytes, plasma cells and foamy macrophages, as well as multifocal to coalescing areas of hemorrhage and fibrin deposition. Intranuclear eosinophilic inclusion bodies were rarely observed in neurons and astrocytes. On leptomeninges, there was diffuse inflammatory infiltrates of lymphocytes and plasma cells. Inflammation was also seen in a milder degree in the spinal cord, characterized by infiltrate of lymphocytes at grey matter, mainly around vessels. A herpesvirus which induced typical cytopathic effect in cell cultures was recovered from tissues. The isolated virus was typed as BoHV-5 by nucleotide sequencing analysis of the gC coding region.Discussion: The diagnosis of meningoencephalomyelitis associated to BoHV-5 was based on epidemiological, clinical, macroscopical, histological, virological and genomic findings. In the outbreak here reported, the disease occurred in young animals, with low morbidity but high lethality rates. Clinical signs on this case were consistent with previous reports on the literature. Bovines affected by BoHV-5 may display no gross lesions within the CNS; however, inflammatory and degenerative changes are frequently seen, characterized by malacia, leptomeningeal vessels hyperemia, edema and hemorrhages. Histologically, non-suppurative necrotizing meningoencephalitis is seen, with perivascular inflammatory infiltrates and, occasionally, intranuclear eosinophilic inclusion bodies in astrocytes and neurons. Similar but milder lesions were seen in the spinal cords of two of the necropsied calves, a feature which has only been previously associated to BoHV-1 infections. The identification of the implicated agent was accomplished by virus isolation in cell cultures followed by genome typing. Specific treatments for this condition are not currently available, and the number of animals that recover from clinically apparent disease is extremely low. Preventive measures are based on serological testing of herds, and segregation or elimination of seropositive calves. To avoid progression of the disease in seropositive animals, control efforts must be directed to avoid stressful conditions. Vaccination with BoHV-1 and BoHV-5 vaccines is expected to confer protection to clinical disease.

PM 7/21 (2) Ralstonia solanacearum, R. pseudosolanacearum and R. syzygii (Ralstonia solanacearum species complex)

Specific scopeThis Standard describes a diagnostic protocol for Ralstonia solanacearum, Ralstonia pseudosolanacearum and Ralstonia syzygii, i.e. phylotype/sequevar strain in the Ralstonia solanacearum Species Complex (RSSC).It should be used in conjunction with PM 7/76 Use of EPPO diagnostic protocols.Specific approval and amendmentApproved in 2003‐09. First revised in 2018‐02.

Sensibility of Spondias purpurea L. (Anacardiaceae) exposed to fluoride-simulated fog

Fluoride is the atmospheric pollutant with the highest phytotoxicity arising from aluminum smelting fertilizers, glass and, ceramic industry. The most affected plant organ is the leaf, in which the fluoride penetrates through two main ways: absorption by epidermis cuticle or absorption by stomata. Species susceptible to fluoride are potential tools in bioindicator studies because they provide a means for detecting the presence of the pollutant in the environment at a low cost. The aim of this study is to evaluate the sensibility of Spondias purpurea L. (Anacardiaceae) to fluoride through the simulated fog. Simulation was performed by applying 500 ml of solution containing 15 mg L−1 of potassium fluoride per plant for 20 min daily and for 10 consecutive days. The data of leaflet abscission, visual damage, and climate conditions were recorded daily. At the end of the experiment, data from the cell death analysis, electrolyte leakage determination, and accumulation of the fluoride in the dried matter were collected. Moreover, samples for phenolic compound accumulation, anatomical, and micromorphological analyses were collected. After 24 h, the appearance of visible damage was observed in the leaves with fluoride treatment. This damage consisted, mostly, of necrosis, chlorosis, and leaf apical shriveling. The necroses manifested in gray and brown discoloration. Leaf abscission was intense in the young leaves. Microscopic damage consisted of protoplast retraction, phenolic compound accumulation, collapse, hyperplasia, and cell rupture. Turgidity loss, epicuticular wax erosion, and damage of stomata and trichomes beyond the presence of fungal hyphae were noted in the epidermis. Evans Blue detected dead cell groups in the transition region between the necrosis and the apparent healthy area. Fluoride affected the selective permeability of the membrane, which was observed in this study according to the high rate of electrolyte leakage. Plants exposed to fluoride accumulated 14.48 times more fluoride in their leaves than plants in the control group. It follows that S. purpurea is susceptible to fluoride and responds quickly to the presence of this pollutant. Marginal and apical necrosis, presence of phenolic compounds, fluoride accumulation, anatomical alterations, and leaflet abscission in the young leaves are biomarkers of fluoride effects in this species. Therefore, S. purpurea is potentially useful in biomonitoring programs.

First report on wilt disease of mango caused by Ceratocystis fimbriata in Uttar Pradesh, India

Wilt disease of mango (Mangifera indica L.) caused by Ceratocystis spp. has caused great losses to mango industry. In India, increased incidence of mango wilt was reported by farmers in Uttar Pradesh. Since, no systematic studies regarding mango wilt were carried out in past, major mango growing areas of Uttar Pradesh were surveyed during 2012–2016 to understand the disease. Incidence of Ceratocystis wilt was confirmed in 42 orchards out of 1917 orchards surveyed. Disease symptoms appeared in the form of sudden wilt, decline and branch drying. Severe oozing of gum was mostly observed from trunk of wilted trees. On exposing vascular tissues, reddish-brown to dark brown or black discoloration was observed. The fungus was isolated from infected root and stem tissues and rhizosphere soil on carrot discs and identified as Ceratocystis fimbriata. Pathogenicity of C. fimbriata was confirmed on mango seedlings. PCR amplification of the ITS regions resulted in amplification of ~ 500 base pair (bp) fragment. The BLASTn sequence analysis showed alignment of identified isolate with C. fimbriata that was clearly delineated from close species viz. C. manginecans and C. albifundus. Thus, this is the first report of C. fimbriata causing mango wilt from India, which is unique and variable from C. maginecans reported earlier from Pakistan and Oman. Ceratocystis wilt disease of mango has now posed great threat to mango industry in Uttar Pradesh, India.

Abstract P3-01-11: Discoloration after injection of super paramagnetic iron oxide (SPIO) for sentinel node biopsy. A long term qualitative follow-up study

Abstract Background SPIO has a similar detection ratio as Technetium99 and Patent Blue to identify sentinel nodes (SN). No allergic reactions have been observed and no nuclear medicine facilities are needed, making logistics easier. At the Academic Hospital in Uppsala, Sweden, SPIO has been used routinely for three years. After breast conserving surgery (BCS) many women developed a brownish discoloration at the injection site. The discoloration stays for a long time. To study the patient-experienced cosmetic discomfort and the natural history of the discoloration we followed our first 153 BCS women for more than two years. After modifying the technique of injection we registered the discoloration in the following 115 women. Methods All women injected with a retro-areolar injection of SPIO between April 2015 and October 2016 were included. The women were telephone interviewed every third month. The size of the discoloration was self-assessed and the cosmetic discomfort was classified by a scale from 0 (no discomfort) to 5 points (very discomforting). Photos were taken in selected cases after 1-2 years. Between November 2016 and April 2017, a deeper, para-tumoral injection was used and discoloration was noted 3 weeks after surgery. Results Ninety of 153 women (58.8%) developed a discoloration after a retro-areolar injection. The mean size was 26.1cm2 (2-100cm2). The discoloration had vanished in 6.8% and 11.4% of the women after 1 and 2 years, respectively. The mean size of the discolorations was 18.2cm2 (1-66cm2) and 13.6cm2 (1-66cm2) after 1 and 2 years. The intensity of the color was continuously fading. The cosmetic discomfort was assessed as 2.2 points after 1 year and 1.0 after 2 years. Of 115 women with a deeper injection, 32.2% developed a discoloration with a mean size of 13.1cm2 (1-36cm2), three weeks after surgery. The incidence and size of the discolorations were statistically significantly less and smaller after a deeper injection (p<0.001 and p=0.001, respectively). SNs were identified in 91% of women with a retro-areolar injection and in 93% of those with a deeper injection. Conclusions After an injection of SPIO, a discoloration might develop and the discoloration can stay for more than 2 years in many women. However, the color fades and the size diminish continuously and the women do not consider the discoloration a major cosmetic problem. A deeper injection reduces the incidence and the size of the discoloration with a similar detection ratio of SN. In our next study, we aim to reduce the volume of SPIO and thereby hopefully reduce the discoloration. Patient-assessed cosmetic discomfort after 1 and 2 yearsPatient age1 year2 years<55 years3.10.9≥55 years1.91.00=no discomfort, 5=very discomforting Citation Format: Wärnberg M, Karakatsanis A, Abdsaleh S, Wärnberg F. Discoloration after injection of super paramagnetic iron oxide (SPIO) for sentinel node biopsy. A long term qualitative follow-up study [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr P3-01-11.

First Report of Pythium myriotylum D., Causing Damping Off and Root Rot in Chili Pepper (Capsicum annum L.) from Punjab, Pakistan

Chilli pepper (Capsicum annuum L.) has significant importance as a cash crop, like other vegetables, and occupies 20% of the total vegetable production area in Pakistan. Pythium spp. are notorious fungal-like agents that may result in 90% plant death either as pre and/or post-emergence damping off. During surveys of nine major chilli growing areas of Punjab, Pakistan in September-November 2015-16, disease incidences ranging from 13.8 to 45.4% was recorded. The symptoms consisted of damping off, and on older seedlings (15 to 30 days post-transplanting) reduced growth, wilting, water-soaking, brown discoloration and root rot. Infected roots were cut into small pieces, surface sterilized using 1% sodium hypochlorite for 3 to 5 min, followed by a rinse with sterile distilled water. Root saps were serially diluted to appropriate dilutions, streaked onto corn meal agar (CMA) medium supplemented with ampicillin (250 mg/l), rifampicin (10 mg/l) and pimaricin (10 mg/l) (Jeffers and Martin 1986) and incubated at 28...