Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241115EnglishN2019March14HealthcareChronic lymphocytic leukemia —Internationa Prognostic Index for Macedonian Patients with Chronic Lymphocytic Leukemia English0105Trajkova S.English Cevreska L.English Ivanovski M.English Popova-Labacevska M.English Pivkova-Veljanovska A.English Panovska-Stavridis I.EnglishIntroduction: Several prognostic factors have been identified to predict the outcome of patients with Chronic lymphocytic leukemia(CLL), but only a few studies investigated more markers together. To predict the time to first treatment (TFT) we integrated the data of traditional staging system, cytogenetic aberrations, and mutational status of immunoglobulin heavy chain variable region (IGHV) in Chronic lymphocytic leukemia– Internationa Prognostic Index(CLL-IPI). Aim of the study: To validate the prognostic value of chronic lymphocytic leukemia-international prognostic index (CLL-IPI) for Macedonian CLL patients. Material and Methods: The study is set up retrospectively and includes 75 patients with CLL diagnosed and treated at the University Clinic of Hematology for a period of time from January 2011 to January 2018. The median follow-up was 36 months (1-72 months).We recognized the prognostic markers of TFT,in line of definition that prognostic markers we incorporated the data of Rai staging system, most adverse cytogenetic marker and mutational status of immunoglobulin heavy chain. Results: The statistical data of the 75 patients shows that58.7% were males and 41.3% were females, with a median age of 64.3 (42-85) years old. The median TFS for low CLL-IPI (n=20), intermediate CLL-IPI (n=27), high risk CLL-IPI (n=15) and very high risk group (n=7) according to the CLL-IPI scoring system was 7.9, 7.6, 7.0 and 5.8 months, respectively. The median OS for low risk group was 58.5 for intermediate, high, and very high risk group was 37.8, 34.6 and 30 months, respectively. The estimated 5-year OS rate was 98.7%, 92%, 40% and 33%, respectively for low risk group intermediate, high, and very high risk group. Multivariate analysis indicated that del 17p (P< 0.00874) was independent prognostic factors of TFS. Conclusions: CLL-IPI is the powerful tool for risk stratification in Macedonian CLL patients and this system also provided treatment recommendations for the different patient risk subgroup.Though, it is unknown how newly developed CLL therapies and newly identified prognostic and predictive markers would be merged into this revised staging system. EnglishLeukemia, Lymphocytic, Chronic, PrognosisINTRODUCTION Chronic lymphocytic leukemia (CLL) is the most common type of adult leukemia in Western countries. In Macedonia, with the aging of the population and the increasingly westernized lifestyle, the incidence rate is increasing year by year. The median survival of CLL patients is approximately 10 years, but the prognosis of different patients is highly heterogeneous. One group  of patients can implement the "watch and waiting" strategy and have no treatment indication for life, but one third of the patients need treatment at the time of diagnosis, and only few patients  turn into invasive lymphoma with a very poor prognosis. The traditional clinical staging systems Rai and Binet, based only on physical examination and laboratory findings do not include new implemented prognostic markers like TP53 deletion and immunoglobulin heavy chain variable region (IGHV) genes. Mutation status of IGHV and molecular markers have strong prognostic significance of CLL.  Understanding the biology of the disease, most patients are at the early stage of diagnosis at the time of diagnosis, so the prognosis of patients with Rai and Binet clinical judgment system has become incomplete. Therefore, a new prognostic system is needed for accurate prognostic stratification of CLL. The international group of researchers conducted a literature search for phase II and phase III clinical trials of CLL published in a pediod of time from January 1. 1950 to December 31. 2010, including eight prospective trials in their analyses. In total, the studies included 3.473 treatment-naïve patients at both early and advanced CLL stages (median age, 61 years; range, 27-86 years) from France, Germany, Poland, the United Kingdom, and the United States. Patients were followed for a median of 80 months (1). The researchers externally validated the model in two additional datasets: a cohort from the Mayo Clinic in Rochester, Minnesota (n=838), and the SCALE Scandinavian population-based case-control study (n=416). Mean patient age in these cohorts was 62 years (range = 25-89 years), and these patients were followed for a median of 63 months (1). From the group of 27 baseline factors the CLL-IPI researchers examined in the training dataset, five emerged as independent prognostic markers for overall survival (OS): TP53 status (no abnormalities vs. del17p, TP53 mutations, or both) IGHV mutational status (mutated vs. unmutated) Serum ß2-microglobulin (B2M) concentration (≤3.5 mg/L vs. >3.5 mg/L) Clinical stage (Binet A or Rai 0 vs. Binet B-C or Rai I-IV) Age (≤65 years vs. >65 years) Each variable was then assigned an individual weight. The researchers then added all of these factors together, derived a prognostic score ranging from 0-10, and identified four risk groups with significantly different rates of OS at five years (pEnglishhttp://ijcrr.com/abstract.php?article_id=2585http://ijcrr.com/article_html.php?did=25851.An international prognostic index for patients with chronic lymphocytic leukaemia (CLL-IPI): a meta-analysis of individual patient data [J]. Lancet Oncol, 2016 , 17 ( 6 ): 779 - 790 . DOI:10.1016/S1470-2045 ( 16) 30029-8 . 2.Hallek M , Cheson BD , Catovsky D , et al .Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on Chronic Lymphocytic Leukemia updating the National Cancer Institute-Working Group 1996 guidelines[J]. Blood, 2008, 111(12):5446-5456.DOI: 10.1182/blood-2007-06-093906 . 3.Wierda WG, O&#39;Brien S, Wang X, Faderl S, Ferrajoli A, Do KA, et al. Prognostic nomogram and index for overall survival in previously untreated patients with chronic lymphocytic leukemia. Blood. 2007; 109(11): 4679–85. 4.  Qin SC , Xia Y , Miao Y , et al .MYD88 mutations predict unfavorable prognosis in chronic lymphocytic leukemia patients with mutated IGHV gene[J]. Blood Cancer J,2017,7(12):651.DOI:10.1038/s41408-017 -0014-y . 5.Hallek M .Chronic lymphocytic leukemia: 2017 Update ON Diagnosis, Risk Stratification, and treatment[J]. Am J Hematol, 2017,92(. 9):946-965.The DOI: 10.1002 / ajh.24826 .  6.Pflug N , Bahlo J , Shanafelt TD , et al .Development of a comprehensive prognostic index for patients with chronic lymphocytic leukemia[J]. Blood,2014,124(1):49-62. DOI: 10.1182/blood-2014-02 -556399 .  7.Rossi D , Rasi S , Spina V , et al .Integrated mutational and cytogenetic analysis identifies new prognostic subgroups in chronic lymphocytic leukemia[J]. Blood, 2013,121(8):1403-1412. DOI:10.1182/blood-2012- 09-458265 . 8.Molica S , Giannarelli D , Levato L , et al .Assessing time to first treatment in early chronic lymphocytic leukemia (CLL): a comparative performance analysis of five prognostic models with inclusion of CLL-international prognostic index (CLL-IPI)[J]. Leuk Lymphoma,2017,58(. 7):1736-1739. DOI: 10.1080 / 10428194.2016.1257791 .  9.Gentile M , Shanafelt TD , Mauro FR , et al .Comparison between the CLL-IPI and the Barcelona-Brno prognostic model: analysis of 1299 newly diagnosed cases[J]. Am J Hematol, 2018, 93(2):E35-35E37.DOI: 10.1002 / ajh.24960 .

Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241115EnglishN2019March14HealthcareThe Myelogram: Its Contribution to the Diagnosis of Hemopathies English0609Boufrioua El GhaliEnglish Belkhair jihaneEnglish Allali AsmaEnglish Mouamin MaryamEnglish Mohamed ChakourEnglishBackground: The myelogram is the collection of elements provided by microscopic examination of a hematopoietic bone marrow smears collected by puncture-suction. It is considered one of the most valuable diagnostic tools for assessing hematological abnormalities. Methods: We conducted the present retrospective study of 305 patients collected over a 3 year period, from February 2015 through June 2018, performed in the hematology laboratory of the Avicenna military hospital in Marrakech to expose the main bone marrow indications, review its diagnostic contribution, evaluate the practice of the myelogram in our context, and propose recommendations to improve the practice of this examination. Results: The myelogram study allowed to make the diagnosis of 53 (17, 4%) megaloblastosis cases, fifty five (18%) cases of medullary plasmocytosis. Acute myeloblastic leukemia was found in 27 patients (8.8%) followed by chronic lymphocytic leukemia in 11 patients (3.6%), comes after chronic myeloid leukemia in 11 cases (4.3%), while acute lymphoblastic leukemia is in 16 cases (5.2%). The myelogram was normal in 60 cases (19.6%). It objectified a reactional marrow in 49 cases (16%). The myelogram was impossible to interpret in 21 cases (7%), in which the sample was diluted. Conclusion: This general review of haemopathies has highlighted the importance of the myelogram as an indispensable tool for diagnosis. In our study it was possible to know the contribution of this examination, which is linked to its good interpretation. EnglishIntroduction  The myelogram is the collection of elements provided by microscopic examination of a hematopoietic bone marrow smears collected by puncture-suction. It includes the assessment of the smear richness in nucleated cells (megakaryocyte lineage included), the percentage of each cell category and their morphology [1]. It also detects the presence of parasites, infectious agents and tumour cells of spinal and extraspinal cord origin. Thus, it is a cytological examination which allows the qualitative and quantitative analysis of cells belonging to the hematopoietic medullary precursor compartment [2]. It is considered one of the most valuable diagnostic tools for assessing hematological abnormalities. Its indications include diagnosis, classification and therapeutic follow-up of hematological diseases [3]. We conducted the present retrospective study to expose the main bone marrow indications, review its diagnostic contribution, evaluate the practice of the myelogram in our context, and propose recommendations to improve the practice of this examination. Patients and Methods This is a retrospective study of a representative series of 305 patients collected over a 3 year period, from February 2015 through June 2018, performed in the hematology laboratory of the Avicenna military hospital in Marrakech. The data collection was conducted using a farm return by excluding extensions assessment and non-hematological spinal disorders. In this study, all patients of both sexes regardless of age or socio-economic level were concerned. All medullary punctures were performed in the departments with the doctors at the bedside of the patient. The location of the sampling was variable, the sternum facing the 2nd intercostal space was the site of choice for adults, the anterior superior and posterior superior iliac spines were for the children. The punctures were made with Mallarmé trocar and colored with May Grunwald Giemsa which is the reference color in hematology, it consists of applying two neutral dyes; the May-Grünwald’s alcoholic solution, and Giemsa’s 10% diluted alcoholic solution. As for the coloration Perls it is performed on spinal cord smears as part of the primary classification of myelodysplastic syndromes, this coloring highlights insoluble cellular complexes containing iron: hemosiderin, Pappenheimer granules (fine iron-rich granules) = overloaded iron mitochondria. Determining Myeloperoxydase activity by testing (MPO) is routinely used for acute leukemia diagnosis. Its positivity represents a strong marker of myeloid differentiation. Data entry and analysis were performed using the Microsoft Office Excel 2016 software for Windows version 10.0. Results The mean age of patients was 42 years, with minimum age of 1 year and maximum age being 70 years. Children of less than 18 years represent 10% of our patients. A light male predominance was noted with a sex-ratio of 1, 2. Fifty three percent of bone marrow smears performed on hospitalized patients in different services, mainly in the service of internal medicine with a rate of 43% and 47% was conducted on not hospitalised patients. Hemogram disturbance was the first indication of myelogram, that is to say 53% of cases, followed by tumor syndrome in 23% of cases, then the suspicion of monoclonal gammapathy 17% of the cases (Table I). Among the disturbances in blood count (CBC) listed, Macrocytic anemia was the most common abnormality with a rate exceeding 19%, followed by normochromic normochromic anemia found in 14% of the cases. Bicytopenia and pancytopenia respectively represent 12% and 6% of all disturbances. Among the 57 of macrocytic anemia cases found in the hemograms performed, 47 had megaloblastosis. Myelograms done within the framework of thrombocytopenia turned out to be normal, except for one patient who was in coma and suffers from thrombocytopenia and leukocytosis and in whom the myelogram showed acute myeloid leukemia. 18 myelograms were conducted on patients suffering from a pancytopenia: 15 had tumor associated syndrome, and their myelograms showed acute myeloid leukemia. Two cases of megaloblastosis, and one showed a dystrophic plasmocytosis. Myelograms of 51 patients showed a bone syndrome with suspicion of gammopathy, had revealed; a multiple myeloma in 42 patients, chronic lymphocytic leukemia (CML) in three patients, 5 cases of reactive plasma cells, and 1 case of plasma leukemia. In the case of splenomegaly; the myelogram objectified three medullary megaloblastosis and chronic lymphocytic leukemia. Four myelograms were made for suspicion of medicinal side effects: two patients showed rheumatoid arthritis under methotrexate, following the discovery of a hemogram pancytopenia, and for which the myelogram revealed a case of megaloblastosis, and a normal marrow in the other case. Two patients with diabetes under sulfonamide who had thrombocytopenia, and for which the myelogram did not objectify abnormalities. Ten myelograms were performed as part of a nephropathy assessment. Medullary plasmocytosis was found in seven cases, a CML in a case and a dystrophic pith in a patient with chronic kidney failure and a pancytopenia. In total, the myelogram study allowed to make the diagnosis of 53 (17, 4%) megaloblastosis case, fifty five (18%) cases of medullary plasmocytosis. Acute myeloblastic leukemia was found in 27 patients (8.8%) followed by chronic lymphocytic leukemia in 11 patients (3.6%), comes after chronic myeloid leukemia in 11 cases (4.3%), while acute lymphoblastic leukemia is in 16 cases (5.2%). The myelogram was normal in 60 cases (19.6%). It objectified a reactional marrow in 49 cases (16%). The myelogram was impossible to interpret in 21 cases (7%), in which the sample was diluted (Table II). Discussion This work focuses on the contribution of the myelogram in the diagnosis of blood diseases, in the hematology laboratory at the Avicenna Marrakech military hospital has retrospectively noted an average prevalence of 101 myelogram / year. This average result is attributed to the participation of the hematology laboratory of the Mohammed VI University Hospital Centre of Marrakech, which supports a significant number of preformed myelograms. The sampling of this exam at University Hospital Centre of Treichville is 275 myelograms per year [1], and 1100 myelograms / year at the central hematology laboratory at Ibn Sina Hospital in Rabat in 2016 [2]. Compared to the hemogram, the myelogram is a relatively infrequent examination. This is due to the speed and ease of the conduct of the hemogram: thanks to automation. As for the myelogram, it also represents a major examination of major utility, but it has few indications compared to hemogram. The average age of patients in our series was 43 years, close to the average ages found in studies in Burkina Faso (42 years old) [4], of Cameroon (44,27) [5]. While the study conducted in Rabat [2] showed an average age of 14 years, this is explained by the predominance of the pediatric population in this series, which is 61%. The male predominance found in our study is comparable to the results reported in the studies of Casablanca (1, 4) [6], Bass-Normandy (1, 13) [7], in Senegal (1, 6) [8], Mali (1, 9) [9] in the Democratic Republic of Congo (1, 7) [10]. Men are more exposed to environmental factors than women because of their professional activity (use of herbicides and pesticides) [11, 12]. The most affected age group was 30 to 40 years old, whereas the study conducted in Rabat [2] found a high prevalence of malignant hemopathies (MH) in the age group 10 to 19, this difference being explained by the fact that that our population was essentially an adult population (90% of the patients were at least 18 years old). We identified 11 main indications of the myelogram in our study; disturbances of the hemogram were the main indication, followed by tumor syndrome, these are the classic indications of the myelogram; Hugard [3], Ouattara [1] found similar results, concerning the frequency of a hemogram anomalies with predominance of pancytopenia in their series. The blood count generally showed bicytopenia (anemia and thrombocytopenia) associated with variable leukocytosis. Anemia and thrombocytopenia were generally associated with leukocytosis in patients with MH in a series in Burkina Faso [13]. These biological signs have also been observed in Tunisia [12] and Cote d&#39;Ivoire [14, 15]. Pancytopenia was also noted in 6% of patients. In fact, the circumstances of discovery of MH, especially leukaemias, are generally signs of bone marrow failure more or less associated with a tumor syndrome [13]. Among the 57 macrocytic anemias, 47 had megaloblastosis with myelogram. This aspect is compatible with vitamin deficiency [16, 17], which have been confirmed by the corresponding vitamin dosages (folic acid and vitamin B12). Lymphoproliferative syndromes are ranked first in the MH study in 24.5% of cases. A prevalence of lymphoproliferative syndromes of 67, 3% was found in Ivory Coast [18]. Multiple myeloma represents 15.5% of the cases in our study, which was close to the 12.2% reported in Abidjan [19], but above the 7% average rates found in Mali and Senegal [9, 8]. The CLL occupied 5.2% of the cases, unlike in Niger where it occupied 33.3%. [20]. Acute leukaemias represent 13.1% (40 cases) of MH. They occupied the third position as nosological group in our study while in Ivory Coast study; they were in second with 21.3% of cases [21]. In the study by Seiter et al [22], acute myeloid leukemia (AML) represents 80% of acute leukemia (AL) in adults and 20% of acute children leukemia. Myeloproliferative syndromes came fourth with 4.5% (14 cases); in comparison with a Cameroonian study [21]. In our study, CML represents 4.2% of cases while other authors found higher proportions: 18.9 in Niger [20] and 9.3 in Ivory Coast [19]. In our series, we had only one patient in whom there was an appearance of hemophagocytosis in the myelogram. Confronted with other clinico-paraclinical data described in the literature [23], the diagnosis of macrophage activation syndrome (MAS) was retained. We have 4 cases presenting different medicinal side effects: Two patients followed for rheumatoid arthritis under methotrexate that had pancytopenia. Pancytopenia under methotrexate at low doses is a rare but serious complication [24]. Its prevention is based on the close supervision of elderly patients, especially in cases of polypharmacy and hypoalbuminemia. The results of myelogram in these patients showed normal marrow in one and megaloblastosis in the other. This effect has been mentioned by several authors [24]. The other two patients with type 2 sulfamide diabetes had thrombopenia: known side effect of this treatment [25]. The myelogram in our 2 patients showed a normal aspect of the marrow which favors the immunological origin [25]. Reaction marrows were noted in 16% of patients in our series. This medullary reaction is not specific and is observed in a large number of pathological states (toxic, viral, parasitic) [25]. Diluted myelogram represented 7% of cases in our series. The most frequent causes of dilution were the technical errors during the sampling, the other causes of diluted marrow being: fibroses and medullary aplasias [26]. Conclusion  On a going back 3 years at Avicenna Military Hospital, this study allowed us to take stock of the hematological conditions diagnosed within our service. This general review of hemopathies has highlighted the importance of the myelogram as an indispensable tool for diagnosis. In our study it was possible to know the contribution of this examination, which is linked to its good interpretation, which is in turn directly related to the rigor of its performance. Highlight the various technical errors, and make recommendations to improve its implementation. Conflict of Interest: The authors declare no potential conflicts of interest, financial or otherwise. Englishhttp://ijcrr.com/abstract.php?article_id=2586http://ijcrr.com/article_html.php?did=2586 Ouattara S, Danho B, Kone M, Tea D. La pratique du myélogramme au CHU de Treichville : bilan de 19 années d’activité. Médecine d&#39;Afrique Noire; 2000:47(5) Omar El hamdaoui. L’apport du myélogramme dans le diagnostic des hémopathies: expérience du laboratoire central d’hématologie de l’hôpital Ibn Sina. Thèse de médecine. universite Mohammed V – rabat. Faculté de médecine et de pharmacie Rabat;2018,22 p. HugardL, Bon J.C, Force -barge P, Porte J, RODIER J. Contribution à l’étude de la prévalence de l’anémie chez l’enfant en milieu tropical. CMS ELF-Congo - Pointe Noire. Méd. Afr. Noire 1997; 44 (1): 40-44. Ouédraogo SM, Hien F, Bazié W, Millogo A, Drabo YJ. Place des hémopathies malignes en service de Médecine interne du CHU SouroSanou (Burkina Faso). Mali Médical 2011 ; XXVI : 17-21. Moueleu N, Ngouadjeu D, Ngo S, Luma H. Épidémiologie des hémopathies malignes recensées en milieu hospitalier au Cameroun. Médecine et Santé Tropicales, Vol. 28, N8 1 - janvier-février-mars 2018 M. Harif, A. Benider, M. Bennani Othmani et al. Registre des cancers de la région du Grand Casablanca. Année 2005, 2006,2007, édition 2012.  www.orsbn.org/publication/bulletin_11.pdf.  Mortalité par hémopathies malignes en Basse-Normandie. Le Bulletin de l’observatoire régional de la santé de Basse-Normandie numéro 11, Décembre 1997. Thiam D, Diop TM, Tallarmin F, Toure AO, Diakhate L. Epidemiology and therapy of malignant hemopathies in Senegal. Hematol Cell Ther 1996 ; 38 : 187-91. Diallo DA, Cissoko LS, Cissoko Y, et al. Epidémiologie actuelle des hémopathies malignes dans les services d’hématologie oncologie médicale et de médecine interne de l’hôpital du Point G, Bamako, Mali. Mali Medical 2005 ; 4 : 1-8. Mufuta NJP, Mbayo K, Kayembe NZ, Gini EK, Mbuyi M. Cytologie des hémopathies malignes dans deux formations médicales de Kinshasa. Ann AfrMed 2013 ; 6. Henry K. Neoplastic disorders of lympho-reticular tissue in Henry K. and Symmers, W St C : Systemic Pathology : thymus, lymph node, node spleen  and lymphatics.3 rd Edition. London : Churchill Livingtone, 1992; p. 611960. Elloumi M, Hafsia R, Omri EL, et al. Caractéristiques épidémiologiques, cliniques et cytohématologiques des leucémies aigués lymphoblastiques de l’adulte en Tunisie. Tunis Me ´dical 2002 ; 80 : 199-202 Ouédraogo SM, Hien F, Bazie W, Millogo A, Drabo YJ. Place des hémopathies malignes en service de Médecine interne du CHU Souro Sanou (Burkina Faso). Mali Médical 2011 ; XXVI : 17-21. Sangaré M, Kassi HY, N’Dogom M, Yayo-Ayé ME, Inwoley AK, Sawadogo D. Pro?l biologique des leucémies aigues non myéloides en Co ˆte d’Ivoire. Rev Int ScMéd 2013 ; 15 : 49-53. Sawadogo D, Lartey MT, Kouassi D, et al. Impact de la chimiothérapie sur les pro?ls hématologiques et biochimiques au cours des hémopathies malignes à Abidjan. Cahier Santé2002 ; 12 : 229-32. Sebahoun G, Sainty D, Horschowski N. Ponction médullaire et biopsie médullaire. Editions techniques. Encycl. Méd. Chir. (Paris, France), Hématologie 13000, A30,1991,7p.                                                  Zittoun J, Zittoun R. Les anémies macrocytaires. Encycl. Méd. Chir. (Paris, France), Sang, 13002,A10,1985,14 p. N’Dhatz Comoe E, Kof? KG, Ayemou R, et al. Prévalence et incidence des hémopathies malignes au CHU de Yopougon. Revintscme ´d 2012 ; 14 : 205-8.   Téa D, Yao T, Abissey A, Anoh J, Rain JD. Panorama des hémopathies diagnostiquées au laboratoire central du CHU de Treichville bilan de 10 années d’activité. Médecine d&#39;Afrique Noire : 39-43. Mounkaila B, Touré I, Gragnic G, MounkailaI. Hémopathies malignes à Niamey   à propos  de 90 observations sur 6ans. Méd Afr Noire 1996; 43: 472-5. N’Dhatz Comoe E, Koffi KG, Ayemou R, et al. Prévalence et incidence des hémopathies malignes au CHU de Yopougon. Rev int sc méd 2012 ; 14 : 205-8. Seiter K, Harris J. E. Myeloid leukemia staging.  N Engl J Acute Med 1999; 341(11):807-813 Henter JI, Horne A, Aricó M, Egeler RM, Filipovich AH, Imashuku S, et al. Diagnostic and therapeutic guidelines for hemophagocytic lymphohistiocytosis. Pediatr Blood Cancer 2007; 48:124-131.  Sallah S, Hanrahan L R,  Phillips D L. Intrathecal methotrexate induced megaloblastic anemia in patients with acute leukemia. Arch Pathol Lab Med. 1999 Sep; 123(9):774-7 Dearden C. Wade R. Else M. Richards S. Milligan D. 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Radiance Research AcademyInternational Journal of Current Research and Review2231-21960975-5241115EnglishN2019March14Life SciencesTwo New Cladosporium Species from Murshidabad District, West Bengal, India English1013Dinesh HaldarEnglishIntroduction: Cladosporium is a member of dematiaceous hyphomycetes which is characterized and easily recognised by its unique structure of the conidiogenous loci and conidial hila. Despite being imperfect in forms, they are the most versatile organisms in nature. While working on dematiaceous hyphomycetes from Murshidabad district, two specimens of Cladosporium had been collected which on critical examination characterised as two new species. Methods: The infected leaves with distinct symptoms were collected and dried to make herbarium specimens. The microscope slides were prepared in lacto-phenol cotton blue mixtures of the recognised species. The fungi, treated here were studied in their natural habitat on the leaves of Clitoria terrnatea and Hibiscus mutabilis. Results and Discussion: Two new species of Cladosporium viz. Cladosporium clitoriae Haldar and Cladosporium murshidabadense Haldar on Clitoria ternatea Land Hibiscus mutabilis L. respectively had been collected, figured and described in the present communication with notes on their phylogeny and ecology. EnglishAnamorphic, Hyphomycetes, Taxonomy, sp.novINTRODUCTION The genus Cladosporium, a well documented fungus and was established by Link  in  the year 1815 with Cladosporium herbarum (Pers: Fr.) Link ex S.F. Gray is the type species. It had been named as “Klados” means a branch, hence branched spore chains. The taxonomic posi­tion of the genus Cladosporium is almost accepted as being a member of the form family Cladosporiaceae under the order Hyphomycetes of the form class Deuteromycetes. The genus Cladosporium is well characterized and easily recognised by its unique structure of the conidiogenous loci and conidial hila which classified as coronate, i.e., composed of a central convex dome surrounded by a raised periclinal rim. Cladosporoid are common and worldwide distribution and it is one of the largest, most heterogeneous genera of hyphomycetes, comprising more than 772 names i.e., valid, invalid, legitimate and illegitimate species, varieties, formae as well as herbarium name. Reasons for this vast number of taxa probably reside in the imprecise, wide circumscription of this genus in literature, the strong morphological variability of most species and the occurrence of some species on a wide range of substrates. The members of Cladosporium is a dematiaceous fungi occurring  on all kinds of living and dead leaves and stems of herbaceous and woody plants, as secondary invaders on necrotic leaf lesions caused by other fungi, are frequently isolated from air, soil, food stuffs, paint, textiles and other organic matters. Reviews keys of Cladosporoid taxa on living plants were provided  by  a large number of worker from India and abroad. Some of them are : Amanelah, Beharvandi & Zafari2, Avasthi, Gautam &Bhadauria3  Bensch et al4 ., Braun &Hill5, Braun &Crous6,  Crous7, Crous et al,8. Crous 9 ,Das10,David &David11,Dugan et al.12,Ellis13, Ellis14, Heuchert, Braun &Schubert15, He & Zhanh16,Jang et al17,Kirk18,  Mu?enko, Schubert & Kozlowska 19, Partridge & Morgan-Jones 20, Schubert  et al,21 Schubert & Braun22,Seifert ,Nickerson  & Corlet23 Schubert & Braun24, Wirsel SGR & Runge- Frobose25, Thomma, Van Esse & Crous26 , Schubert et al 27 , Braun & Schubert28, Zhang et al29, Sharma et al30, Sandoval-Denis et al31, Subramania32, Kamal33 and Haldar34. During working on the foliicolous fungi of Murshidabad district in the year 2016 of West Bengal the author had collected two mem­bers of Hyphomycetes growing on the living leaves of Clitoria ternatea L. (Fabaceae)and Hibiscus mutabilis L.(Malvaceae). which on critical examination found to be two new species of the genus Cladospori­um. Hence, these two species. Cladosporium clitoriae  Haldar sp.nov. and Cladosporium murshidabadense Haldar sp.nov. have been created as new taxa (Myco Bank35). The district of Murshidabad is located in the central part of the state of West Bengal under Presidency Division, lies  between 23°43/ to24052/ N latitude and 87049/ to 88044/ E longitude and occupies an area of 5341sq km. The district is separated from the Maldah district by the river Ganga on its north. The eastern boundary of the district is formed by Bangladesh. It is bounded by the districts of Burdwan and Nadia on the south, the Birbhum district and the state of Jharkhand located in the western side. The river Bhagirathi flowing across the district divides it into almost two equal parts. The western part is known as Rarh which is substantially continuation of sub-vindhyan region. The soil of this district is primarily alluvial. The tract east to river is known as Bagri and is covered with recent alluvium consisting sandy clay. The elevation of the district is from 10m to 50m above MSL. Ganga and its tributaries such as Bhagirathi, Jalangi and Bhairab are the important rivers of the district. The district is oppressively having hot summer, short winter, high humidity and good rainfall during monsoon season. The average annual rainfall of the district is 1400 mm. MATERIALS AND METHODS Specimens with typical disease symptoms of Cladosporoid fungi  on living leaves were collected during field trips. The infected leaves were detached intact from the host plants and they were kept in polythene bags and processed by following standard techniques Castaneda85. Pho­tographs of the infected spots on the host leaves were cap­tured by Sony DSC-HX200, camera and for the examination of fungal structure and spore morphology, the microscope slides were prepared in lacto-phenol cotton blue mixtures of the recognised species. Morphotaxonomic study of the associated fungi was done through the low and high magnification 100x400 of the compound microscope, (Olympus-CX21i FS1 Research Microscope) by using USB INSTA CMOS camera. The mi­crophotographs were stored in electronic format JPEG. Holotypes being deposited at AMH, Agharkar Research Institute (ARI), Pune (MS), India and isotypes retained in the Departmental her­barium (KNC)for future reference. RESULTS Cladosporium clitoriae Haldar sp.nov.(Fig.1) Myco Bank MB 829285  Incidence in winter, infection foliicolous inciting whitish, necrotic,  spots amphigenous distinct on upper surface, numerous, subcircular to irregular, sometimes surrounded by grey margin, lesions occasionally developing shot-hole appearance, 1.5-2.5 in diam. caespituli epiphyllous, brown, mycelium immersed, superficial, smooth. stroma amphigenous,well developed, arising from the epidermal hairs of the host tissue, composed of thick walled isodiametric  type of cells, light brown: conidiophores solitary to fasciculate, with 2-8 divergent stalks in a fascicle arising from the base of the stroma and rarely through the stomata, straight to curved, pale brown, septate, simple to branched, sometimes nodose with terminal and intercalary swelling, thick walled, smooth with distinct scars, after succession of conidia, apex sub acute to obtuse, average length 277.56-627.92 µm  and breadth-29.16µm.conidia solitary to catenate up to 3 in a chain, pale olivaceous, usually straight, rarely curved, cylindrical, ellipsoidal, thin walled, smooth to verruculose, up to 4 septate, often with distinct protuberant scar at  both ends, average length 53.49-212.89 µm and breadth 29.16 µm. Etymology: Referring to the genus of the host. Habitat in folis vivis Clitoria ternatea L.(Fabaceae),Ring Road, Berhampore, Murshidabad, West Bengal, India, AMH 9847(Holotypus), KNC 142 (Isotypus). D.Haldar, 06.11.1016. Cladosporium murshidabadense sp.nov.(Fig.2) MycoBank MB 829286 Incidence in rainy season, spots formed on lamina, amphigenous, semi circular to circular, dark brown centre surrounded by blackish margin, sometimes no definite spots, only greyish discoloration without any boundary,0.5—10 mm in diam,; caespituli amphigenous ,vein limited, unevenly distributed over the spots, black; stroma present(23.60-297.87 µm),occasionally only a few brownish spherical to gobular hyphal cells, substomatal; conidiophores amphigenous, solitary to fasciculate, in fascicles  2-7 stalks emerging the stomatal opening, straight to flexus, brown to pale brownish, simple or branched, thick walled, smooth, very often nodose, plurisepted (1-9 septa),septa distinct, often geniculate, often terminal vesicular swelling with distinct spore scar lying at the apex or by the side wall of the conidiophores,(average length 559.33-1124.60 µm) and breadth(40.79-48.02 µm);conidia solitary to catenate, cylindrical, ellipsoidal, lemon shaped, oblong, pale brown, smooth, thick walled 0-3 septate, hilum distinct aat one end or both ends, average length,(53.26-137.97 µm) and average breadth (25.81-54.88 µm). Specimen studied : On Hibiscus mutabilis L. (Malvaceae), Kossim Bazar, Berhampore, Murshidabad, West Bengal, India, AMH-9748(Holotypus), KNC 317(Isotypus), D.Haldar,2 June,2016. Etymology: Referring to the genus of the host. No species of Cladosporium has yet been recorded and described on the hosts Clitoria ternatea L.(Fabaceae) and Hibiscus mutabilis L.(Malvaceae) and there is no record of this genus so far on other members of the family Fabaceae and Malvaceae respectively. So, the present fungus is described and illustrated here as a new taxon. DISCUSSION The fungi Cladosporium clitoriae Haldar and Cladosporium murshidabadense Haldar has been found to occur profusely during winter months to spring and early summer. The host plants Clitoria ternatea L. and Hibiscus mutabilis L. both are economically important. The roots of Clitoria ternatea L. is used as powerful cathartic and diuretic. Flowers yield a blue dye. Seeds contain a fixed oil. Both seeds and root-bark contain tannin. Purgative and leaves used as fodder. On the other hand Hiiscus mutabilis L. bark yields a strong fibre of inferior quality. Leaves used for cough, menorrhagia, dysuria and wounds caused by burns and scalds. Flowers given in pectoral and   pulmonary complaints36. CONCLUSION Two new species of Cladosporium  viz   Cladosporium clitoriae Haldar and Cladosporium murshidabadense Haldar  growing on two ornamental plants namely Clitoria ternatea L. and Hibiscus mutabilis L. They cause severe leaf infection of the host plants resulted which the growth of the plants become stunted. A pathologist should develop the control measures of the disease. ACKNOWLEDGEMENTS The author is thankful to the Principal, Krishnath College, Murshidabad, West Bengal for rendering help during the pre­sent work.. The author expresses his sincere gratitude to the Curator of AMH-ARI, Pune for depositing holotype of the specimens and providing the accession numbers. I wish to acknowledge the extended help of Dr. S. 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