Prevalence of Anaplasma marginale in different tick species from Ngorongoro Crater, Tanzania.

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Fyumagwa, Robert D; Simmler, Pascale; Meli, Marina L; Hoare, Richard; Hofmann-Lehmann, Regina; Lutz, Hans

2009-04-06

In 2001, Ngorongoro Crater was infested with high density of ticks on grassland, livestock and wildlife which was also associated with high mortality. Adult ticks were collected, identified, processed for nucleic acids extraction and a molecular analysis was performed to determine the range of tick species harboring Anaplasma marginale. The real-time PCR was used in the amplification of rickettsia DNA in tick pools (n=527) from 11 identified tick species. Six tick species were detected with A. marginale DNA including Amblyomma gemma, Rhipicephalus appendiculatus, R. compositus, R.decoloratus, R. praetextatus and R. pulchellus. The detection rate in each tick species was 3%, 0.7%, 2%, 13%, 1.8%, and 6.2%, respectively. Five of the positive tick species excluding R.decoloratus have previously not been described to transmit A. marginale. High diversity of tick species detected with A. marginale in Ngorongoro Crater is likely to increase a risk to susceptible animals of contracting the infection.

Immune responses in cattle vaccinated with gamma-irradiated Anaplasma marginale

International Nuclear Information System (INIS)

Sharma, S.P.; Bansal, G.C.

1986-01-01

The infectivity and immunogenecity of gamma-irradiated Anaplasma marginale organisms were studied in bovine calves. The severity of Anaplasma infection based on per cent infected red blood cells, haematological values and mortality was more in animals immunized with blood exposed to 60 kR in comparison to those inoculated with blood irradiated at 70, 80 and 90 kR. The immunizing controls demonstrated a significantly high parasitaemia, marked anaemia and more deaths. Marked and prolonged cell-mediated and humoral immune responses detectable in the first 3 weeks of post-immunization may be responsible for conferring of protective immunity. (author)

Anaplasma Marginale isolation from infected bovine erythrocytes or from its floating culture

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Canon Q, Y.

1986-01-01

Isolation of Anaplasma Marginale is of great importance because this is the cause of anaplasmosis in cattle. Anaplasmosis is a mortal disease and spreads easily. To isolate the anaplasm, four different experiments were developed; in the first experiment, the parasite was isolated from parasitic blood, by means of three methods; osmotic shock, sonic vibration and treatment with hemolisine. In the second experiment, the parasite source was the top of the bacteriologic culture of anaplasma marginale obtained by means of slow centrifugation. In the third experiment, it was used parasitic blood diluted in PBS and liquid nitrogen criopreserved. In the fourth experiment.it was used parasitic blood which was separated by means of sonic oscillation. This method was more adequate to free the parasite from the host cell. Differential centrifugation was the best method to separate parasite of the stroma and ghost cells

Identification of Anaplasma marginale type IV secretion system effector proteins.

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Svetlana Lockwood

Full Text Available Anaplasma marginale, an obligate intracellular alphaproteobacterium in the order Rickettsiales, is a tick-borne pathogen and the leading cause of anaplasmosis in cattle worldwide. Complete genome sequencing of A. marginale revealed that it has a type IV secretion system (T4SS. The T4SS is one of seven known types of secretion systems utilized by bacteria, with the type III and IV secretion systems particularly prevalent among pathogenic Gram-negative bacteria. The T4SS is predicted to play an important role in the invasion and pathogenesis of A. marginale by translocating effector proteins across its membrane into eukaryotic target cells. However, T4SS effector proteins have not been identified and tested in the laboratory until now.By combining computational methods with phylogenetic analysis and sequence identity searches, we identified a subset of potential T4SS effectors in A. marginale strain St. Maries and chose six for laboratory testing. Four (AM185, AM470, AM705 [AnkA], and AM1141 of these six proteins were translocated in a T4SS-dependent manner using Legionella pneumophila as a reporter system.The algorithm employed to find T4SS effector proteins in A. marginale identified four such proteins that were verified by laboratory testing. L. pneumophila was shown to work as a model system for A. marginale and thus can be used as a screening tool for A. marginale effector proteins. The first T4SS effector proteins for A. marginale have been identified in this work.

Anaplasma marginale major surface protein 2 CD4+-T-cell epitopes are evenly distributed in conserved and hypervariable regions (HVR), whereas linear B-cell epitopes are predominantly located in the HVR.

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Abbott, Jeffrey R; Palmer, Guy H; Howard, Chris J; Hope, Jayne C; Brown, Wendy C

2004-12-01

Organisms in the genus Anaplasma express an immunodominant major surface protein 2 (MSP2), composed of a central hypervariable region (HVR) flanked by highly conserved regions. Throughout Anaplasma marginale infection, recombination results in the sequential appearance of novel MSP2 variants and subsequent control of rickettsemia by the immune response, leading to persistent infection. To determine whether immune evasion and selection for variant organisms is associated with a predominant response against HVR epitopes, T-cell and linear B-cell epitopes were localized by measuring peripheral blood gamma interferon-secreting cells, proliferation, and antibody binding to 27 overlapping peptides spanning MSP2 in 16 cattle. Similar numbers of MSP2-specific CD4(+) T-cell epitopes eliciting responses of similar magnitude were found in conserved and hypervariable regions. T-cell epitope clusters recognized by the majority of animals were identified in the HVR (amino acids [aa] 171 to 229) and conserved regions (aa 101 to 170 and 272 to 361). In contrast, linear B-cell epitopes were concentrated in the HVR, residing within hydrophilic sequences. The pattern of recognition of epitope clusters by T cells and of HVR epitopes by B cells is consistent with the influence of protein structure on epitope recognition.

Knockout of an outer membrane protein operon of anaplasma marginale by transposon mutagenesis

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Large amounts of data generated by genomics, transcriptomics and proteomics technologies have increased our understanding of the biology of Anaplasma marginale. However, these data have also led to new assumptions that require testing, ideally through classic genetic mutation. One example is the def...

Prevalence of Babesia spp. and Anaplasma marginale in cattle in the municipality of Palma, MG

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Marluce Aparecida Mattos Paula

2015-12-01

Full Text Available ABSTRACT. Paula M.A.M., Oliveira F.C.R., Melo Jr O.A. & Frazão-Teixeira E. [Prevalence of Babesia spp. and Anaplasma marginale in cattle in the municipality of Palma, MG.] Prevalência de Babesia spp. e Anaplasma marginale em bovinos no município de Palma, MG. Revista Brasileira de Medicina Veterinária, 37(4: 359-365, 2015. Laboratório de Biologia Estrutural, Instituto Oswaldo Cruz/ Fiocruz, Avenida Brasil, 4365, Manguinhos, Rio de Janeiro, RJ 21040-361, Brasil. E-mail: edwards.teixeira@ioc.fiocruz.br We verified the prevalence of hemoparasites in 40 cattle with ages varying from one month to 12 years old, in two farms of the Municipality of Palma, Minas Gerais state, Brazil. Two blood smear samples were collected from each animal: one from the tail tip and another from the ear tip. The smears were fixed, stained and observed under 100X lighted microscope magnifying glass. Twenty- -seven out of 40 animals studied (67.5% had at least one species of hemoparasite. Among these, 21 (52.5% were infected with Babesia spp., 10 (25% with Anaplasma marginale and four (10% parasitized with both hemoparasites. The studied region is potentially enzootic for the detected parasites and there is high risk for clinical cases of tick-borne disease. Both anatomic points, tail and ear tips, are good spots for blood collection and smear confection for hemoparasite investigation.

Transplacental transmission of Anaplasma marginale in beef cattle chronically infected in southern Brazil

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Hermann Eduardo Gonzalez Grau

Full Text Available In this study, we have investigated the incidence of transplacental transmission of Anaplasma marginale in chronically infected cows with no history of acute anaplasmosis during gestation. In addition, we evaluated various techniques for detection of transplacental transmission ofA. marginale. Blood samples were collected from 30 cows at three different periods: at the time of artificial insemination, at gestational diagnosis, and after calving. Also, blood was collected from the newborn calves, including one sample before colostrum intake, and another three days after birth. A. marginale-specific antibodies were detected in 100% of the cows with an indirect fluorescent antibody test (IFAT, and in 97% of them, using an indirect enzyme-linked immunosorbent assay (ELISA. Also, we observed that all of the three-day-old newborn calves were seropositive by IFAT. According to polymerase chain reaction, 63.3% of the cows were carriers of A. marginale, as well as 6.7% of the newborn calves. This represented a transplacental transmission rate of 10.5%. Furthermore, a correlation of 93.3% was observed between the two serodiagnostic techniques, demonstrating that both ELISA and IFAT can be used in epidemiological surveys of A. marginale. These results confirm the occurrence of transplacental transmission of A. marginale in chronically infected cows and suggest the importance of this transmission route in areas of enzootic instability.

Frequency of antibodies to Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax and Borrelia burdgorferi in cattle from the northeastern region of the state of Pará, Brazil Freqüência de anticorpos para Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax e Borrelia burgdorferi em bovinos do nordeste do Estado do Pará, Brasil

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Daniel S. Guedes Junior

2008-06-01

Full Text Available Babesiosis, anaplasmosis, and trypanosomosis are relevant diseases, potentially causing morbidity in cattle, leading to economic losses. Borreliosis is import as a potential zoonosis. The objective of this study was to determine, by indirect enzyme-linked immunosorbent assay (ELISA, the frequency of seropositive cattle to Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax and Borrelia burgdorferi in cattle from the Northeastern region of Pará, Brazil. Sera samples from 246 female adult cattle from municipalities of Castanhal and São Miguel do Guamá were used. Crude antigens ELISAs were used to detect antibodies to all agents, except to A. marginale, to which an indirect ELISA with recombinant major surface 1a protein (MSP1a antigen was used. Overall frequencies of seropositive animals were: B. bigemina - 99.2%; B. bovis - 98.8%; A. marginale - 68.3%; T. vivax - 93.1% and B. burgdorferi - 54.9%. The frequencies of seropositive cattle to B. bovis and B. bigemina suggest a high rate of transmission of these organisms by tick in the studied region, which can be classified as enzootically stable to these hemoprotozoans. The low frequency of seropositive cattle to A. marginale may be attributed to a lower sensitivity of the recombinant antigen ELISA utilized or a distinct rate of inoculation of this rickettsia by ticks, as compared with Babesia sp. transmission. The high frequency of seropositive cattle to T. vivax indicates that this hemoprotozoan is prevalent in herds from the Northeastern region of Pará. The rate of animal that showed homologues antibodies to B. burgdorferi indicates the presence of the tickborne spirochaetal agent in the cattle population in the studied region.A babesiose, a anaplasmose e a tripanossomose são enfermidades relevantes, potencialmente causadoras de morbidade em bovinos, levando a perdas econômicas. A borreliose assume importância como zoonose potencial. O objetivo desse estudo foi determinar

Diversity of ankA and msp4 genes of Anaplasma phagocytophilum in Slovenia.

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Strašek Smrdel, Katja; von Loewenich, Friederike D; Petrovec, Miroslav; Avšič Županc, Tatjana

2015-03-01

Granulocytic anaplasmosis is a tick transmitted emerging disease in Europe and worldwide. The agent, Anaplasma phagocytophilum is transmitted by ticks of the genus Ixodes and causes infections in humans and domestic animals. The analysis of different target genes showed that in nature several genetic variants of A. phagocytophilum were present. The purpose of our study was to genetically characterize A. phagocytophilum strains from eight humans, 16 dogs, 12 wild boars, one bear and 18 tick pools from Slovenia. Therefore, the ankA and msp4 genes of A. phagocytophilum were chosen. The same genetic ankA and msp4 variant of A. phagocytophilum was detected in humans, wild boar and a part of the pooled ticks indicating that it circulates in a zoonotic cycle between wild boar and ticks. In dogs, three ankA variants of A. phagocytophilum were detected. One of them was identical to the one that was found in humans. In contrast, all dogs harboured the same msp4 variant as humans and wild boar. In ticks, numerous ankA and msp4 variants were present. Copyright © 2014 Elsevier GmbH. All rights reserved.

Spatio-temporal variations and genetic diversity of Anaplasma spp. in cattle from the North of Tunisia.

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Belkahia, Hanène; Ben Said, Mourad; El Mabrouk, Narjesse; Saidani, Mariem; Cherni, Chayma; Ben Hassen, Mariem; Bouattour, Ali; Messadi, Lilia

2017-09-01

In cattle, anaplasmosis is a tick-borne rickettsial disease caused by Anaplasma marginale, A. centrale, A. phagocytophilum, and A. bovis. To date, no information concerning the seasonal dynamics of single and/or mixed infections by different Anaplasma species in bovines are available in Tunisia. In this work, a total of 1035 blood bovine samples were collected in spring (n=367), summer (n=248), autumn (n=244) and winter (n=176) from five different governorates belonging to three bioclimatic zones from the North of Tunisia. Molecular survey of A. marginale, A. centrale and A. bovis in cattle showed that average prevalence rates were 4.7% (minimum 4.1% in autumn and maximum 5.6% in summer), 7% (minimum 3.9% in winter and maximum 10.7% in autumn) and 4.9% (minimum 2.7% in spring and maximum 7.3% in summer), respectively. A. phagocytophilum was not detected in all investigated cattle. Seasonal variations of Anaplasma spp. infection and co-infection rates in overall and/or according to each bioclimatic area were recorded. Molecular characterization of A. marginale msp4 gene indicated a high sequence homology of revealed strains with A. marginale sequences from African countries. Alignment of 16S rRNA A. centrale sequences showed that Tunisian strains were identical to the vaccine strain from several sub-Saharan African and European countries. The comparison of the 16S rRNA sequences of A. bovis variants showed a perfect homology between Tunisian variants isolated from cattle, goats and sheep. These present data are essential to estimate the risk of bovine anaplasmosis in order to develop integrated control policies against multi-species pathogen communities, infecting humans and different animal species, in the country. Copyright © 2017 Elsevier B.V. All rights reserved.

Nanoparticle-Based Delivery of Anaplasma marginale Membrane Proteins; VirB9-1 and VirB10 Produced in the Pichia pastoris Expression System.

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Zhang, Bing; Cavallaro, Antonio S; Mody, Karishma T; Zhang, Jun; Deringer, James R; Brown, Wendy C; Mahony, Timothy J; Yu, Chengzhong; Mitter, Neena

2016-11-05

Bovine anaplasmosis or cattle-tick fever is a tick-borne haemolytic disease caused by the rickettsial haemoparasite Anaplasma marginale in tropical and subtropical areas of the world. While difficult to express, the proteins VirB9-1 and VirB10 are immunogenic components of the outer membrane type IV secretion system that have been identified as candidate antigens for vaccines targeting of A. marginale . Soluble VirB9-1 and VirB10 were successfully expressed using Pichia pastoris . When formulated with the self-adjuvanting silica vesicles, SV-100 (diameter: 50 nm, and pore entrance size: 6 nm), 200 µg of VirB9-1 and VirB10 were adsorbed per milligram of nanoparticle. The VirB9-1 and VirB10, SV-100 formulations were shown to induce higher antibody responses in mice compared to the QuilA formulations. Moreover, intracellular staining of selected cytokines demonstrated that both VirB9-1 and VirB10 formulations induced cell-mediated immune responses in mice. Importantly, the SV-100 VirB9-1 and VirB10 complexes were shown to specifically stimulate bovine T-cell linages derived from calves immunised with A. marginale outer membrane fractions, suggesting formulations will be useful for bovine immunisation and protection studies. Overall this study demonstrates the potential of self-adjuvanting silica vesicle formulations to address current deficiencies in vaccine delivery applications.

Molecular Epidemiological Survey and Genetic Characterization of Anaplasma Species in Mongolian Livestock.

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Ochirkhuu, Nyamsuren; Konnai, Satoru; Odbileg, Raadan; Murata, Shiro; Ohashi, Kazuhiko

2017-08-01

Anaplasma species are obligate intracellular rickettsial pathogens that cause great economic loss to the animal industry. Few studies on Anaplasma infections in Mongolian livestock have been conducted. This study examined the prevalence of Anaplasma marginale, Anaplasma ovis, Anaplasma phagocytophilum, and Anaplasma bovis by polymerase chain reaction assay in 928 blood samples collected from native cattle and dairy cattle (Bos taurus), yaks (Bos grunniens), sheep (Ovis aries), and goats (Capra aegagrus hircus) in four provinces of Ulaanbaatar city in Mongolia. We genetically characterized positive samples through sequencing analysis based on the heat-shock protein groEL, major surface protein 4 (msp4), and 16S rRNA genes. Only A. ovis was detected in Mongolian livestock (cattle, yaks, sheep, and goats), with 413 animals (44.5%) positive for groEL and 308 animals (33.2%) positive for msp4 genes. In the phylogenetic tree, we separated A. ovis sequences into two distinct clusters based on the groEL gene. One cluster comprised sequences derived mainly from sheep and goats, which was similar to that in A. ovis isolates from other countries. The other divergent cluster comprised sequences derived from cattle and yaks and appeared to be newly branched from that in previously published single isolates in Mongolian cattle. In addition, the msp4 gene of A. ovis using same and different samples with groEL gene of the pathogen demonstrated that all sequences derived from all animal species, except for three sequences derived from cattle and yak, were clustered together, and were identical or similar to those in isolates from other countries. We used 16S rRNA gene sequences to investigate the genetically divergent A. ovis and identified high homology of 99.3-100%. However, the sequences derived from cattle did not match those derived from sheep and goats. The results of this study on the prevalence and molecular characterization of A. ovis in Mongolian livestock can facilitate

Nanoparticle-Based Delivery of Anaplasma marginale Membrane Proteins; VirB9-1 and VirB10 Produced in the Pichia pastoris Expression System

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Bing Zhang

2016-11-01

Full Text Available Bovine anaplasmosis or cattle-tick fever is a tick-borne haemolytic disease caused by the rickettsial haemoparasite Anaplasma marginale in tropical and subtropical areas of the world. While difficult to express, the proteins VirB9-1 and VirB10 are immunogenic components of the outer membrane type IV secretion system that have been identified as candidate antigens for vaccines targeting of A. marginale. Soluble VirB9-1 and VirB10 were successfully expressed using Pichia pastoris. When formulated with the self-adjuvanting silica vesicles, SV-100 (diameter: 50 nm, and pore entrance size: 6 nm, 200 µg of VirB9-1 and VirB10 were adsorbed per milligram of nanoparticle. The VirB9-1 and VirB10, SV-100 formulations were shown to induce higher antibody responses in mice compared to the QuilA formulations. Moreover, intracellular staining of selected cytokines demonstrated that both VirB9-1 and VirB10 formulations induced cell-mediated immune responses in mice. Importantly, the SV-100 VirB9-1 and VirB10 complexes were shown to specifically stimulate bovine T-cell linages derived from calves immunised with A. marginale outer membrane fractions, suggesting formulations will be useful for bovine immunisation and protection studies. Overall this study demonstrates the potential of self-adjuvanting silica vesicle formulations to address current deficiencies in vaccine delivery applications.

Uso de inóculo padronizado de Anaplasma marginale e da quimioprofilaxia no controle da anaplasmose bovina

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Ribeiro M.F.B.

2003-01-01

Full Text Available O uso de inóculo homólogo padronizado de Anaplasma marginale foi comparado à prática de quimioprofilaxia com diidrato de oxitetraciclina na redução da riquetsemia e do volume globular (VG causada pela anaplasmose bovina. Os animais que receberam o inóculo (10(7 hemácias com Anaplasma marginaleapresentaram, ao serem desafiados em campo, riquetsemia média de 1,2% e redução média de VG de 23,0%, significativamente inferiores às do grupo-controle (P<0,05. No experimento de quimioprofilaxia os animais que receberam três doses de diidrato de oxitetraciclina (20mg/kg, com intervalos de 25 dias, apresentaram riquetsemia de 2,7% e redução de 36,3% no VG, significativamente inferiores às do grupo-controle (P<0,05. Ambas as medidas preventivas testadas foram eficientes na redução da intensidade da riquetsemia e na queda do VG.

Transmisión de Anaplasma marginale por garrapatas

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Kelly A. Brayton

2012-01-01

Full Text Available Anaplasma marginale, patógeno de distribución mundial, es transmitido por garrapatas Ixódidas. Comprender su complejo desarrollo dentro de la garrapata vector, permitirá la predicción de brotes y ofrecerá oportunidades para controlar su transmisión. En este trabajo se revisa su ciclo básico de desarrollo junto con los estudios recientes acerca de las diferencias de transmisión entre cepas, que delinean aspectos de la interacción patógeno - vector. Bacterias, virus o protozoarios transmitidos por artrópodos causan enfermedades severas, tanto en humanos como en animales. Las enfermedades infecciosas transmitidas por garrapatas, entre las que incluimos a la Anaplasmosis (A. marginale, babesiosis (Babesia bigemina, B. bovis, B. divergens y Theileriosis (Theileria annulata, T. parva, se encuentran entre las más importantes en el ámbito mundial, con pérdidas cercanas a los siete mil millones de dólares anualmente; y, a pesar de su impacto, permanecen escasamente bajo control, basado primordialmente en la aplicación de acaricidas, para interrumpir su transmisión. La aparición de garrapatas resistentes a múltiples sustancias acaricidas, representa una amenaza en este tipo de control y, como resultado, hay un resurgimiento de la investigación para el desarrollo de nuevas estrategias para su control. Nuevas opciones para prevenir la transmisión de patógenos de animales por garrapatas, será el resultado de entender las interacciones garrapata patógeno; proceso que culmina con el desarrollo de la infección y transmisión exitosa. En todos los casos de patógenos transmitidos por garrapatas, el desarrollo de la infección se realiza coordinamente a los momentos de adhesión y alimentación del vector sobre el animal. Esto sucede por la interdependencia en la señalización entre el patógeno y el vector al alimentarse y, por ello, será susceptible de intervención.

Molecular detection and characterization of Babesia bovis, Babesia bigemina, Theileria species and Anaplasma marginale isolated from cattle in Kenya.

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Adjou Moumouni, Paul Franck; Aboge, Gabriel Oluga; Terkawi, Mohamad Alaa; Masatani, Tatsunori; Cao, Shinuo; Kamyingkird, Ketsarin; Jirapattharasate, Charoonluk; Zhou, Mo; Wang, Guanbo; Liu, Mingming; Iguchi, Aiko; Vudriko, Patrick; Ybanez, Adrian Patalinghug; Inokuma, Hisashi; Shirafuji-Umemiya, Rika; Suzuki, Hiroshi; Xuan, Xuenan

2015-09-30

Infections with Babesia bovis, Babesia bigemina, Theileria species and Anaplasma marginale are endemic in Kenya yet there is a lack of adequate information on their genotypes. This study established the genetic diversities of the above tick-borne hemoparasites infecting cattle in Kenya. Nested PCR and sequencing were used to determine the prevalence and genetic diversity of the above parasites in 192 cattle blood samples collected from Ngong and Machakos farms. B. bovis spherical body protein 4, B. bigemina rhoptry-associated protein 1a, A. marginale major surface protein 5, Theileria spp. 18S rRNA, T. parva p104 and T. orientalis major piroplasm surface protein were used as the marker genes. B. bovis, B. bigemina, T. parva, T. velifera, T. taurotragi, T. mutans and A. marginale were prevalent in both farms, whereas T. ovis, Theileria sp. (buffalo) and T. orientalis were found only in Ngong farm. Co-infections were observed in more than 50 % of positive samples in both farms. Babesia parasites and A. marginale sequences were highly conserved while T. parva and T. orientalis were polymorphic. Cattle-derived T. parva was detected in Machakos farm. However, cattle and buffalo-derived Theileria were detected in Ngong farm suggesting interactions between cattle and wild buffaloes. Generally, the pathogens detected in Kenya were genetically related to the other African isolates but different from the isolates in other continents. The current findings reaffirm the endemicity and co-infection of cattle with tick-borne hemoparasites, and the role of wildlife in pathogens transmission and population genetics in Kenya.

Eficácia do dipropionato de imidocarb, da enrofloxacina e do cloridrato de oxitetraciclina no tratamento de bovinos naturalmente infectados por Anaplasma marginale

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L.R. Alberton

2015-08-01

Full Text Available O agente de maior importância, em relação à anaplasmose bovina, é o Anaplasma marginale. Os principais sinais clínicos dessa enfermidade são anemia hemolítica, icterícia, dispneia, taquicardia, febre, fadiga, lacrimejamento, sialorreia, micção frequente, anorexia, perda de peso, aborto e morte. A terapia antimicrobiana é o principal protocolo terapêutico. O objetivo do presente trabalho foi avaliar a eficácia do dipropionato de imidocarb, da enrofloxacina e do cloridrato de oxitetraciclina no tratamento de bovinos leiteiros naturalmente infectados por Anaplasma marginale. Para isso, foram avaliados 48 zebuínos mestiços que apresentavam os sinais clínicos sugestivos da doença. Os animais foram submetidos à coleta de sangue para a realização de hemograma e à extração de DNA para a confirmação da presença de A. marginale, por meio da reação em cadeia pela polimerase (PCR. Os animais foram divididos em três grupos experimentais, para realização dos protocolos terapêuticos, utilizando-se dipropionato de imidocarb, oxitetraciclina e enrofloxacina. Trinta e seis animais (75% apresentaram reação positiva ao PCR. Os animais positivos não apresentaram diferenças significativas quanto ao hemograma e ao leucograma quando comparados com os negativos, no entanto os níveis de proteínas séricas foram inferiores nos animais positivos (P<0,05. Os três protocolos terapêuticos foram capazes de reduzir a infecção ao longo do tratamento (P<0,01, porém, após cinco dias de tratamento, a enrofloxacina apresentou maior efetividade em relação aos demais (P<0,01. Após o final do tratamento, nenhum protocolo foi capaz de eliminar totalmente a infecção pelo A. marginale em bovinos naturalmente infectados e manejados a campo.

Anaplasma phagocytophilum MSP4 and HSP70 Proteins Are Involved in Interactions with Host Cells during Pathogen Infection

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Marinela Contreras

2017-07-01

Full Text Available Anaplasma phagocytophilum transmembrane and surface proteins play a role during infection and multiplication in host neutrophils and tick vector cells. Recently, A. phagocytophilum Major surface protein 4 (MSP4 and Heat shock protein 70 (HSP70 were shown to be localized on the bacterial membrane, with a possible role during pathogen infection in ticks. In this study, we hypothesized that A. phagocytophilum MSP4 and HSP70 have similar functions in tick-pathogen and host-pathogen interactions. To address this hypothesis, herein we characterized the role of these bacterial proteins in interaction and infection of vertebrate host cells. The results showed that A. phagocytophilum MSP4 and HSP70 are involved in host-pathogen interactions, with a role for HSP70 during pathogen infection. The analysis of the potential protective capacity of MSP4 and MSP4-HSP70 antigens in immunized sheep showed that MSP4-HSP70 was only partially protective against pathogen infection. This limited protection may be associated with several factors, including the recognition of non-protective epitopes by IgG in immunized lambs. Nevertheless, these antigens may be combined with other candidate protective antigens for the development of vaccines for the control of human and animal granulocytic anaplasmosis. Focusing on the characterization of host protective immune mechanisms and protein-protein interactions at the host-pathogen interface may lead to the discovery and design of new effective protective antigens.

Validation of an improved anaplasma antibody cELISA kit for detection of anaplasma ovis antibody in domestic sheep at the U.S. Sheep Experiment Station in Dubois, ID

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An accurate and simple-to-perform new version of a competitive ELISA (cELISA) kit that became commercially available in 2015 for testing of cattle for antibody to Anaplasma marginale was validated for detection of Anaplasma ovis antibody in domestic sheep. True positives and negatives were identifie...

Effect of gamma irradiation on anaplasma marginale in bovines

International Nuclear Information System (INIS)

Romero, H.Q.; Fernandez, M.V.G. de; Meza, B.R.; Rodriguez, E.

1981-01-01

Because the costs of treatment of anaplasmosis are very high, it was considered of great usefulness to obtain the modification of the pathogenecity and the antigenecity of A. marginale by means of gamma radiation ( 60 Co) to obtain a vaccine. This would be reflected favorably in saving lives as well as to obviate losses of meat and dairy production and the elimination of the risks derived from the introduction of selected cattle into the tropics. The main purpose of this study was to determine the optimal modification of the pathogenecity of A. marginale in splenectomized calves and adult cows, among 4 different radiation doses. (author)

Subdominant Outer Membrane Antigens in Anaplasma marginale: Conservation, Antigenicity, and Protective Capacity Using Recombinant Protein.

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Deirdre R Ducken

Full Text Available Anaplasma marginale is a tick-borne rickettsial pathogen of cattle with a worldwide distribution. Currently a safe and efficacious vaccine is unavailable. Outer membrane protein (OMP extracts or a defined surface protein complex reproducibly induce protective immunity. However, there are several knowledge gaps limiting progress in vaccine development. First, are these OMPs conserved among the diversity of A. marginale strains circulating in endemic regions? Second, are the most highly conserved outer membrane proteins in the immunogens recognized by immunized and protected animals? Lastly, can this subset of OMPs recognized by antibody from protected vaccinates and conserved among strains recapitulate the protection of outer membrane vaccines? To address the first goal, genes encoding OMPs AM202, AM368, AM854, AM936, AM1041, and AM1096, major subdominant components of the outer membrane, were cloned and sequenced from geographically diverse strains and isolates. AM202, AM936, AM854, and AM1096 share 99.9 to 100% amino acid identity. AM1041 has 97.1 to 100% and AM368 has 98.3 to 99.9% amino acid identity. While all four of the most highly conserved OMPs were recognized by IgG from animals immunized with outer membranes, linked surface protein complexes, or unlinked surface protein complexes and shown to be protected from challenge, the highest titers and consistent recognition among vaccinates were to AM854 and AM936. Consequently, animals were immunized with recombinant AM854 and AM936 and challenged. Recombinant vaccinates and purified outer membrane vaccinates had similar IgG and IgG2 responses to both proteins. However, the recombinant vaccinates developed higher bacteremia after challenge as compared to adjuvant-only controls and outer membrane vaccinates. These results provide the first evidence that vaccination with specific antigens may exacerbate disease. Progressing from the protective capacity of outer membrane formulations to

Anaplasma ovis genetic diversity detected by major surface protein 1a and its prevalence in small ruminants.

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Aktas, Munir; Özübek, Sezayi

2018-04-01

Anaplasma ovis is a widely distributed tick-borne rickettsial pathogen of sheep, goats, and wild ruminants. The aims of this study were to assess the prevalence, associations of Anaplasma ovis in sheep and goats, as well as its genetic diversity based on analysis of the msp1α gene. A total of 416 DNA samples from sheep (n = 236) and goats (n = 180) from four provinces in southeastern Turkey were analyzed by PCR. The overall A. ovis prevalence was 18% (CI 14.4-22.1). The infection rates of A. ovis varied from 15.9% to 21.8% in sampled provinces, and they were not significantly different. There was no difference between Anaplasma ovis infection in sheep (20.3%, CI 15.4-26.0) and goats (15.0%, CI 10.1-21.1) or in infection rate of animals 1 year (16.4%, CI 12.4-21.2). A significant association between A. ovis infection and the presence of Rhipicephalus bursa and Rhipicephalus turanicus was observed (P diversity of A. ovis were found in small ruminants in Turkey. Copyright © 2018 Elsevier B.V. All rights reserved.

Prevalencia y detección por PCR anidada de Anaplasma marginale en bovinos y garrapatas en la zona central del Litoral ecuatoriano

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Ariel Escobar Troya

2015-09-01

Full Text Available La bacteria que provoca la anaplasmosis en bovinos se conoce como Anaplasma marginale, es potencialmente transmitida de forma biológica por garrapatas, moscas y fómites o mediante sangre infectada como consecuencia del uso incorrecto de herramientas quirúrgicas. Hasta la fecha, Ecuador carece de estudios actualizados sobre procedimientos eficientes para el diagnóstico específico y, erradicación de A. marginale a través del control de estos insectos, por lo que resulta de gran importancia desarrollar e implementar trabajos relacionados con el uso de esta herramienta molecular, para la detección de la bacteria en vectores de transmisión que provocan la enfermedad en bovinos. En este trabajo, se extrajo ADN eficazmente con el método de Salting Out. Un total de 255 muestras fueron analizadas por PCR anidada, distribuidas del siguiente modo|108| Riphicephalus (Boophilus microplus, |85| bovinos, de estos el 13.46% y 85.48% dieron positivas para la rickettsia, las muestras de Amblyomma spp. |62| todas fueron negativas. El índice de concordancia Kappa |total de bovinos infestados frente a Riphicephalus (Boophilus microplus|, no fue significativo (0.28. Subsiguientemente, se determinó por χ2 (p=0.66 que la presencia o ausencia de la enfermedad es independiente del lugar de donde proviene el bovino.

Molecular detection of tick-borne bacteria and protozoa in cervids and wild boars from Portugal.

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Pereira, André; Parreira, Ricardo; Nunes, Mónica; Casadinho, Afonso; Vieira, Maria Luísa; Campino, Lenea; Maia, Carla

2016-05-10

Wildlife can act as reservoir of different tick-borne pathogens, such as bacteria, parasites and viruses. The aim of the present study was to assess the presence of tick-borne bacteria and protozoa with veterinary and zoonotic importance in cervids and wild boars from the Centre and South of Portugal. One hundred and forty one blood samples from free-ranging ungulates including 73 red deer (Cervus elaphus), 65 wild boars (Sus scrofa) and three fallow deer (Dama dama) were tested for the presence of Anaplasma marginale/A. ovis, A. phagocytophilum, Anaplasma/Ehrlichia spp., Babesia/Theileria spp., Borrelia burgdorferi (sensu lato) (s.l.), and Rickettsia spp. DNA by PCR. Anaplasma spp. DNA was detected in 33 (43.4 %) cervids (31 red deer and two fallow deer) and in two (3.1 %) wild boars while Theileria spp. were found in 34 (44.7 %) cervids (32 red deer and two fallow deer) and in three (4.6 %) wild boar blood samples. Sequence analysis of msp4 sequences identified A. marginale, A. ovis, while the analysis of rDNA sequence data disclosed the presence of A. platys and A. phagocytophilum and T. capreoli and Theileria sp. OT3. Anaplasma spp./Theileria spp. mixed infections were found in 17 cervids (22.4 %) and in two wild boars (3.1 %). All samples were negative for Babesia sp., B. burgdorferi (s.l.), Ehrlichia sp. or Rickettsia sp. This is the first detection of Anaplasma marginale, A. ovis, A. phagocytophilum, A. platys, Theileria capreoli and Theileria sp. OT3 in cervids and wild boars from Portugal. Further studies concerning the potential pathogenicity of the different species of  Anaplasma and Theileria infecting wild ungulates, the identification of their vector range, and their putative infectivity to domestic livestock and humans should be undertaken.

High Prevalence of Anaplasma spp. in Small Ruminants in Morocco.

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Ait Lbacha, H; Alali, S; Zouagui, Z; El Mamoun, L; Rhalem, A; Petit, E; Haddad, N; Gandoin, C; Boulouis, H-J; Maillard, R

2017-02-01

The prevalence of infection by Anaplasma spp. (including Anaplasma phagocytophilum) was determined using blood smear microscopy and PCR through screening of small ruminant blood samples collected from seven regions of Morocco. Co-infections of Anaplasma spp., Babesia spp, Theileria spp. and Mycoplasma spp. were investigated and risk factors for Anaplasma spp. infection assessed. A total of 422 small ruminant blood samples were randomly collected from 70 flocks. Individual animal (breed, age, tick burden and previous treatment) and flock data (GPS coordinate of farm, size of flock and livestock production system) were collected. Upon examination of blood smears, 375 blood samples (88.9%) were found to contain Anaplasma-like erythrocytic inclusion bodies. Upon screening with a large spectrum PCR targeting the Anaplasma 16S rRNA region, 303 (71%) samples were found to be positive. All 303 samples screened with the A. phagocytophilum-specific PCR, which targets the msp2 region, were found to be negative. Differences in prevalence were found to be statistically significant with regard to region, altitude, flock size, livestock production system, grazing system, presence of clinical cases and application of tick and tick-borne diseases prophylactic measures. Kappa analysis revealed a poor concordance between microscopy and PCR (k = 0.14). Agreement with PCR is improved by considering microscopy and packed cell volume (PCV) in parallel. The prevalence of double infections was found to be 1.7, 2.5 and 24% for Anaplasma-Babesia, Anaplasma-Mycoplasma and Anaplasma-Theileria, respectively. Co-infection with three or more haemoparasites was found in 1.6% of animals examined. In conclusion, we demonstrate the high burden of anaplasmosis in small ruminants in Morocco and the high prevalence of co-infections of tick-borne diseases. There is an urgent need to improve the control of this neglected group of diseases. © 2015 Blackwell Verlag GmbH.

Molecular biological identification of Babesia, Theileria, and Anaplasma species in cattle in Egypt using PCR assays, gene sequence analysis and a novel DNA microarray.

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El-Ashker, Maged; Hotzel, Helmut; Gwida, Mayada; El-Beskawy, Mohamed; Silaghi, Cornelia; Tomaso, Herbert

2015-01-30

In this preliminary study, a novel DNA microarray system was tested for the diagnosis of bovine piroplasmosis and anaplasmosis in comparison with microscopy and PCR assay results. In the Dakahlia Governorate, Egypt, 164 cattle were investigated for the presence of piroplasms and Anaplasma species. All investigated cattle were clinically examined. Blood samples were screened for the presence of blood parasites using microscopy and PCR assays. Seventy-one animals were acutely ill, whereas 93 were apparently healthy. In acutely ill cattle, Babesia/Theileria species (n=11) and Anaplasma marginale (n=10) were detected. Mixed infections with Babesia/Theileria spp. and A. marginale were present in two further cases. A. marginale infections were also detected in apparently healthy subjects (n=23). The results of PCR assays were confirmed by DNA sequencing. All samples that were positive by PCR for Babesia/Theileria spp. gave also positive results in the microarray analysis. The microarray chips identified Babesia bovis (n=12) and Babesia bigemina (n=2). Cattle with babesiosis were likely to have hemoglobinuria and nervous signs when compared to those with anaplasmosis that frequently had bloody feces. We conclude that clinical examination in combination with microscopy are still very useful in diagnosing acute cases of babesiosis and anaplasmosis, but a combination of molecular biological diagnostic assays will detect even asymptomatic carriers. In perspective, parallel detection of Babesia/Theileria spp. and A. marginale infections using a single microarray system will be a valuable improvement. Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

Caracterização da imunogenicidade das proteínas recombinantes Virb9, Virb10 e fator termo instável de Elongação de Peptídeos de Anaplasma marginale em camundongos

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Ana Beatriz Canevari Castelão

2012-10-01

Full Text Available Considerando as limitações dos atuais métodos de controle contra a anaplasmose bovina, o desenvolvimento de uma vacina efetiva se faz necessário. A partir do advento da análise genômica e proteômica, novas proteínas de membrana de Anaplasma marginale foram identificadas como possíveis candidatas a componentes de uma vacina, tais como, VirB9,VirB10 e Fator Termo Instavél de Elongação de Peptídeos (EF-Tu. Embora estas proteínas ainda não estejam bem caracterizadas na membrana de A. marginale, a produção destas na forma recombinante (rVirB 9, rVirB10 e rEF-Tutem sido realizada, mas as mesmas ainda não foram exploradas em formulações vacinais. Neste trabalho, avaliou-se ouso de rVirB9, rVirB10 e rEF-Tu emulsionadas em adjuvante Montanide em camundongos. Nas condições testadas, verificou-se a indução de forte resposta imune humoral com a produção de IgG1 e IgG2a, sendo que as proporções dos níveis de produção destas subclasses indicam predomínio de IgG1. Entretanto, esplenócitos de animais, que foram injetados com rVirB9 ou rVirB10, produziram interferon-gama acima do limite de detecção do ensaio após estimulação in vitro, sinalizando assim resposta celular específica. Assim, novas avaliações serão realizadas com a finalidade de modular o perfil de resposta imune obtido em bovinos e avaliar a proteção contra A. marginale.

NCBI nr-aa BLAST: CBRC-ACAR-01-1026 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-ACAR-01-1026 ref|YP_153799.1| hypothetical protein AM518 [Anaplasma marginale ...str. St. Maries] gb|AAV86544.1| hypothetical protein AM518 [Anaplasma marginale str. St. Maries] YP_153799.1 3.7 24% ...

NCBI nr-aa BLAST: CBRC-LAFR-01-0905 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-LAFR-01-0905 ref|YP_153919.1| hypothetical protein AM688 [Anaplasma marginale ...str. St. Maries] gb|AAV86664.1| hypothetical protein AM688 [Anaplasma marginale str. St. Maries] YP_153919.1 5e-08 62% ...

Anaplasma spp. in dogs and owners in north-western Morocco.

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Elhamiani Khatat, Sarah; Daminet, Sylvie; Kachani, Malika; Leutenegger, Christian M; Duchateau, Luc; El Amri, Hamid; Hing, Mony; Azrib, Rahma; Sahibi, Hamid

2017-04-24

Anaplasma phagocytophilum is an emerging tick-borne zoonotic pathogen of increased interest worldwide which has been detected in northern Africa. Anaplasma platys is also present in this region and could possibly have a zoonotic potential. However, only one recent article reports on the human esposure to A. phagocytophilum in Morocco and no data are available on canine exposure to both bacteria. Therefore, we conducted a cross-sectional epidemiological study aiming to assess both canine and human exposure to Anaplasma spp. in Morocco. A total of 425 dogs (95 urban, 160 rural and 175 working dogs) and 11 dog owners were sampled from four cities of Morocco. Canine blood samples were screened for Anaplasma spp. antibodies by an enzyme-linked immunosorbent assay (ELISA) and for A. phagocytophilum and A. platys DNA by a real-time polymerase chain reaction (RT-PCR) targeting the msp2 gene. Human sera were tested for specific A. phagocytophilum immunoglobulin G (IgG) using a commercial immunofluorescence assay (IFA) kit. Anaplasma spp. antibodies and A. platys DNA were detected in 21.9 and 7.5% of the dogs, respectively. Anaplasma phagocytophilum DNA was not amplified. Anaplasma platys DNA was significantly more frequently amplified for working dogs. No statistically significant differences in the prevalence of Anaplasma spp. antibodies or A. platys DNA detection were observed between sexes, age classes or in relation to exposure to ticks. A total of 348 Rhipicephalus sanguineus (sensu lato) ticks were removed from 35 urban and working dogs. The majority of dog owners (7/10) were seroreactive to A. phagoyctophilum IgG (one sample was excluded because of hemolysis). This study demonstrates the occurrence of Anaplasma spp. exposure and A. platys infection in dogs, and A. phagocytophilum exposure in humans in Morocco.

Msp1 Is a Membrane Protein Dislocase for Tail-Anchored Proteins.

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Wohlever, Matthew L; Mateja, Agnieszka; McGilvray, Philip T; Day, Kasey J; Keenan, Robert J

2017-07-20

Mislocalized tail-anchored (TA) proteins of the outer mitochondrial membrane are cleared by a newly identified quality control pathway involving the conserved eukaryotic protein Msp1 (ATAD1 in humans). Msp1 is a transmembrane AAA-ATPase, but its role in TA protein clearance is not known. Here, using purified components reconstituted into proteoliposomes, we show that Msp1 is both necessary and sufficient to drive the ATP-dependent extraction of TA proteins from the membrane. A crystal structure of the Msp1 cytosolic region modeled into a ring hexamer suggests that active Msp1 contains a conserved membrane-facing surface adjacent to a central pore. Structure-guided mutagenesis of the pore residues shows that they are critical for TA protein extraction in vitro and for functional complementation of an msp1 deletion in yeast. Together, these data provide a molecular framework for Msp1-dependent extraction of mislocalized TA proteins from the outer mitochondrial membrane. Copyright © 2017 Elsevier Inc. All rights reserved.

Anaplasma marginale attenuated by irradiation, its potential use as vaccine

International Nuclear Information System (INIS)

Gil, A.; Pupiales, N.; Castro, J.; Higuera, B.; Orrego, J.

1986-01-01

Five groups of 5 calves each were inoculated: group 1 (control) with 5 x 10 8 non-irradiated parasites (day 0) and group 2 with the same dose of irradiated (900 Gy) parasites on day 0. Both groups showed clinical signs of the infection, although group 2 was less severely affected. On day 41 group 3 was inoculated with blood (1 x 10 9 parasites) from an animal in group 2. These parasites were irradiated again (900 Gy) prior to inoculation. On day 59 group 4 was inoculated with blood (1 x 10 9 parasites) from the same animal in group 2. This inoculum was irradiated a second time (900 Gy) and the cells were washed and culture ''IN VITRO'' for 4 days prior to inoculation. Parasitaemia increased abruptly in group 3 (peak = 25%) concomitant with a sudden drop in hematocrit. Two animals died, the others required treatment. Group 4 had a transient and low parasitaemia (peak = 2%) and normal hematocrit values. On day 201 all animals including a new control (group 5) of 5 animals, were inoculated with 2 x 10 9 heterologous parasites. All 5 control animals reached 20% parasitaemia and averaged a -40% hematocrit decrease when treated. In contrast, the remaining animals had from -20% to normal hematocrit values and from 5.6% to non-detectable parasitaemias. All inoculated calves showed high antibody titers which increased once the parasitaemia was overcome, and after challenge. It appears that two (900 Gy) irradiations effectively attenuated A. marginale, conferring an active and long lasting protection. However, inoculum cells had to be washed and cultured to prevent hemolitic shock

First report of Anaplasma ovis in pupal and adult Melophagus ovinus (sheep ked) collected in South Xinjiang, China.

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Zhao, Li; He, Bo; Li, Kai-Rui; Li, Fei; Zhang, Lu-Yao; Li, Xian-Qiang; Liu, Yong-Hong

2018-04-19

Melophagus ovinus (sheep ked) is a blood-feeding ectoparasite that belongs to the family Hippoboscidae (Diptera: Hippoboscoidea) and mainly parasitizes sheep. The life-cycle of M. ovinus consists of three stages: larva, pupa and adult. It has a worldwide distribution and has been found in four provinces of China, especially South Xinjiang. In addition to causing direct damage to animal hosts, M. ovinus serves as a vector for disease transmission. In this study, our aim was to investigate the presence of Anaplasma spp. in pupal and adult M. ovinus. A total of 93 specimens (including eight pupal specimens) of M. ovinus collected in South Xinjiang were selected for isolation of genomic DNA, followed by PCR amplification and sequencing of the msp4 gene of Anaplasma spp. The sequences were analyzed in MEGA 7.0 software and via online BLAST. PCR and sequencing results showed that all the specimens collected in 2013 were free of Anaplasma spp., whereas three and 25 specimens (including five pupal specimens) collected in 2016 and 2017, respectively, tested positive for Anaplasma spp. The analysis of 24 msp4 gene sequences (from four pupal specimens) confirmed the presence of A. ovis in M. ovinus specimens collected in South Xinjiang, China. The detected A. ovis isolates belong to Genotypes II and III. To the best of our knowledge, this is the first report of the detection of A. ovis DNA in pupal M. ovinus, confirming the vertical transmission of A. ovis in M. ovinus and the potential of M. ovinus to serve as a vector for A. ovis.

Genetic diversity of the msp-1, msp-2, and glurp genes of Plasmodium falciparum isolates along the Thai-Myanmar borders.

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Congpuong, Kanungnit; Sukaram, Rungniran; Prompan, Yuparat; Dornae, Aibteesam

2014-08-01

To study the genetic diversity at the msp-1, msp-2, and glurp genes of Plasmodium falciparum (P. falciparum) isolates from 3 endemic areas in Thailand: Tak, Kanchanaburi and Ranong provinces. A total of 144 P. falciparum isolates collected prior to treatment during January, 2012 to June, 2013 were genotyped. DNA was extracted; allele frequency and diversity of msp-1, msp-2, and glurp genes were investigated by nested polymerase chain reaction. P. falciparum isolates in this study had high rate of multiple genotypes infection (96.5%) with an overall mean multiplicity of infection of 3.21. The distribution of allelic families of msp-1 was significantly different among isolates from Tak, Kanchanaburi, and Ranong but not for the msp-2. K1 and MAD20 were the predominant allelic families at the msp-1 gene, whereas alleles belonging to 3D7 were more frequent at the msp-2 gene. The glurp gene had the least diverse alleles. Population structure of P. falciparum isolates from Tak and Ranong was quite similar as revealed by the presence of similar proportions of MAD20 and K1 alleles at msp-1 loci, 3D7 and FC27 alleles at msp-2 loci as well as comparable mean MOI. Isolates from Kanchanaburi had different structures; the most prevalent alleles were K1 and RO33. The present study shows that P. falciparum isolates from Tak and Ranong provinces had similar allelic pattern of msp-1 and msp-2 and diversity but different from Kanchanaburi isolates. These allelic variant profiles are valuable baseline data for future epidemiological study of malaria transmission and for continued monitoring of polymorphisms associated with antimalarial drug resistance in these areas.

Detection of Anaplasma phagocytophilum DNA in Ixodes ticks (Acari: Ixodidae) from Madeira Island and Setubal District, mainland Portugal.

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Santos, Ana Sofia; Santos-Silva, Maria Margarida; Almeida, Victor Carlos; Bacellar, Fátima; Dumler, John Stephen

2004-09-01

A total of 278 Ixodes ticks, collected from Madeira Island and Setubal District, mainland Portugal, were examined by polymerase chain reaction (PCR) for the presence of Anaplasma phagocytophilum. Six (4%) of 142 Ixodes ricinus nymphs collected in Madeira Island and 1 nymph and 1 male (2%) of 93 I. ventalloi collected in Setubal District tested positive for A. phagocytophilum msp2 genes or rrs. Infection was not detected among 43 I. ricinus on mainland Portugal. All PCR products were confirmed by nucleotide sequencing to be identical or to be most closely related to A. phagocytophilum. To our knowledge, this is the first evidence of A. phagocytophilum in ticks from Setubal District, mainland Portugal, and the first documentation of Anaplasma infection in I. ventalloi. Moreover, these findings confirm the persistence of A. phagocytophilum in Madeira Island's I. ricinus.

Detection and phylogenetic characterisation of novel Anaplasma and Ehrlichia species in Amblyomma triguttatum subsp. from four allopatric populations in Australia.

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Gofton, Alexander W; Waudby, Helen P; Petit, Sophie; Greay, Telleasha L; Ryan, Una M; Irwin, Peter J

2017-08-01

Anaplasma and Ehrlichia spp. are tick-borne pathogens that can cause severe disease in domestic animals, and several species are responsible for emerging zoonoses in the northern hemisphere. Until recently, the only members of these genera reported in Australia (A. marginale, A. centrale, and A. platys) were introduced from other continents, through the importation of domestic animals and their associated ticks. However, unique Anaplasma and Ehrlichia 16S rRNA gene sequences were recently detected for the first time in native Australian ticks, particularly in Amblyomma triguttatum subsp. ticks from southwest Western Australia (WA). We used molecular techniques to survey Am. triguttatum subsp. ticks from four allopatric populations in southern and western Australia for Anaplasma and Ehrlichia species, and described here the phylogeny of these novel organisms. An A. bovis variant (genotype Y11) was detected in ticks from two study sites; Yanchep National Park (12/280, 4.3%) and Barrow Island (1/69, 1.4%). Phylogenetic analysis of 16S rRNA and groEL gene sequences concluded that A. bovis genotype Y11 is a unique genetic variant, distinct from other A. bovis isolates worldwide. Additionally, a novel Ehrlichia species was detected in Am. triguttatum subsp. from three of the four study sites; Yanchep National Park (18/280, 6.4%), Bungendore Park (8/46, 17.4%), and Innes National Park (9/214, 4.2%), but not from Barrow Island. Phylogenetic analysis of 16S, groEL, gltA, and map1 gene sequences revealed that this Ehrlichia sp. is most closely related to, but clearly distinct from, E. ruminantium and Ehrlichia sp. Panola Mountain. We propose to designate this new species 'Candidatus Ehrlichia occidentalis'. Anaplasma bovis genotype Y11 and 'Candidatus E. occidentalis' are the first Anaplasma and Ehrlichia species to be recorded in native Australian ticks. Copyright © 2017 Elsevier GmbH. All rights reserved.

Detection of Anaplasma phagocytophilum DNA in Ixodes Ticks (Acari: Ixodidae) from Madeira Island and Setúbal District, Mainland Portugal

Science.gov (United States)

Santos-Silva, Maria Margarida; Almeida, Victor Carlos; Bacellar, Fátima; Dumler, John Stephen

2004-01-01

A total of 278 Ixodes ticks, collected from Madeira Island and Setúbal District, mainland Portugal, were examined by polymerase chain reaction (PCR) for the presence of Anaplasma phagocytophilum. Six (4%) of 142 Ixodes ricinus nymphs collected in Madeira Island and 1 nymph and 1 male (2%) of 93 I. ventalloi collected in Setúbal District tested positive for A. phagocytophilum msp2 genes or rrs. Infection was not detected among 43 I. ricinus on mainland Portugal. All PCR products were confirmed by nucleotide sequencing to be identical or to be most closely related to A. phagocytophilum. To our knowledge, this is the first evidence of A. phagocytophilum in ticks from Setúbal District, mainland Portugal, and the first documentation of Anaplasma infection in I. ventalloi. Moreover, these findings confirm the persistence of A. phagocytophilum in Madeira Island's I. ricinus. PMID:15498168

RINT-1 interacts with MSP58 within nucleoli and plays a role in ribosomal gene transcription

International Nuclear Information System (INIS)

Yang, Chuan-Pin; Kuo, Yu-Liang; Lee, Yi-Chao; Lee, Kuen-Haur; Chiang, Chi-Wu; Wang, Ju-Ming; Hsu, Che-Chia

2016-01-01

The nucleolus is the cellular site of ribosomal (r)DNA transcription and ribosome biogenesis. The 58-kDa microspherule protein (MSP58) is a nucleolar protein involved in rDNA transcription and cell proliferation. However, regulation of MSP58-mediated rDNA transcription remains unknown. Using a yeast two-hybrid system with MSP58 as bait, we isolated complementary (c)DNA encoding Rad50-interacting protein 1 (RINT-1), as a MSP58-binding protein. RINT-1 was implicated in the cell cycle checkpoint, membrane trafficking, Golgi apparatus and centrosome dynamic integrity, and telomere length control. Both in vitro and in vivo interaction assays showed that MSP58 directly interacts with RINT-1. Interestingly, microscopic studies revealed the co-localization of MSP58, RINT-1, and the upstream binding factor (UBF), a rRNA transcription factor, in the nucleolus. We showed that ectopic expression of MSP58 or RINT-1 resulted in decreased rRNA expression and rDNA promoter activity, whereas knockdown of MSP58 or RINT-1 by siRNA exerted the opposite effect. Coexpression of MSP58 and RINT-1 robustly decreased rRNA synthesis compared to overexpression of either protein alone, whereas depletion of RINT-1 from MSP58-transfected cells enhanced rRNA synthesis. We also found that MSP58, RINT-1, and the UBF were associated with the rDNA promoter using a chromatin immunoprecipitation assay. Because aberrant ribosome biogenesis contributes to neoplastic transformation, our results revealed a novel protein complex involved in the regulation of rRNA gene expression, suggesting a role for MSP58 and RINT-1 in cancer development. - Highlights: • RINT-1 is a novel MSP58-interacting protein. • RINT-1 is a nucleolar protein that suppresses ribosomal RNA gene transcription. • RINT-1 and MSP58 cooperate to suppress ribosomal RNA gene transcription. • RINT-1, MSP58, and UBF form a complex on the rDNA promoter.

RINT-1 interacts with MSP58 within nucleoli and plays a role in ribosomal gene transcription.

Science.gov (United States)

Yang, Chuan-Pin; Kuo, Yu-Liang; Lee, Yi-Chao; Lee, Kuen-Haur; Chiang, Chi-Wu; Wang, Ju-Ming; Hsu, Che-Chia; Chang, Wen-Chang; Lin, Ding-Yen

2016-09-16

The nucleolus is the cellular site of ribosomal (r)DNA transcription and ribosome biogenesis. The 58-kDa microspherule protein (MSP58) is a nucleolar protein involved in rDNA transcription and cell proliferation. However, regulation of MSP58-mediated rDNA transcription remains unknown. Using a yeast two-hybrid system with MSP58 as bait, we isolated complementary (c)DNA encoding Rad50-interacting protein 1 (RINT-1), as a MSP58-binding protein. RINT-1 was implicated in the cell cycle checkpoint, membrane trafficking, Golgi apparatus and centrosome dynamic integrity, and telomere length control. Both in vitro and in vivo interaction assays showed that MSP58 directly interacts with RINT-1. Interestingly, microscopic studies revealed the co-localization of MSP58, RINT-1, and the upstream binding factor (UBF), a rRNA transcription factor, in the nucleolus. We showed that ectopic expression of MSP58 or RINT-1 resulted in decreased rRNA expression and rDNA promoter activity, whereas knockdown of MSP58 or RINT-1 by siRNA exerted the opposite effect. Coexpression of MSP58 and RINT-1 robustly decreased rRNA synthesis compared to overexpression of either protein alone, whereas depletion of RINT-1 from MSP58-transfected cells enhanced rRNA synthesis. We also found that MSP58, RINT-1, and the UBF were associated with the rDNA promoter using a chromatin immunoprecipitation assay. Because aberrant ribosome biogenesis contributes to neoplastic transformation, our results revealed a novel protein complex involved in the regulation of rRNA gene expression, suggesting a role for MSP58 and RINT-1 in cancer development. Copyright © 2016 Elsevier Inc. All rights reserved.

First confirmed report of outbreak of theileriosis/anaplasmosis in a cattle farm in Henan, China.

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Cui, Yanyan; Wang, Xiaoxing; Zhang, Yan; Yan, Yaqun; Dong, Haiju; Jian, Fuchun; Shi, Ke; Zhang, Longxian; Wang, Rongjun; Ning, Changshen

2018-01-01

Tick-borne diseases (TBDs) impose a significant constraint to livestock production world widely. In this paper, we presented a case of TBD in a cattle farm in Henan, China. 35 blood samples (7 samples sent by veterinarian, 28 samples gathered by our colleagues) were collected from ill, surviving and asymptomatic cattle and microscopic observation and PCR assays were conducted to characterize the pathogens. Genus Ixodes feeding on these cattle were collected and identified. Theileria annulata-like and Anaplasma marginale-like pathogens were observed in the blood smears stained with Giemsa staining under microscope. Furthermore, 5 out of 7 cattle blood samples were found to be positive for T. annulata by PCR. In the 28 blood specimens, three were positive for T. annulata, while A. marginale DNA was detected in nine blood DNA samples. Besides, 56 ticks feeding on cattle were collected from this farm and were all identified as Rhipisephalus microplus, meanwhile, 10 of them were found to be positive for A. marginale. In addition, phylogenetic analysis of the msp4 gene sequences of A. marginale obtained in this study showed that the isolate from cattle (KX840009) fell in the same clade with that of R. microplus (KX904527), sharing 100% similarity. To the best of our knowledge, this is the first confirmed report of outbreak of theileriosis/anaplasmosis in cattle farms in Henan, China. Copyright © 2017. Published by Elsevier B.V.

Ligasi dan Transformasi Gen MSP1 Plasmodium falciparum Penyebab Malaria di Kota Jayapura

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Arsyam Mawardi

2017-12-01

MSP1 is the most antigenic and expressed protein on merozoite surface when it infects the erythrocytes of malaria patients which leads to its use for vaccine therapy design development. The ligation and transformation process of the MSP1 gene is a gene duplication attempt for producing  the same product during expression. This study aimed to clone P. falciparum MSP-1 gene from tropical malaria patients in Jayapura using pJET1.2/blunt vectors and E. coli DH5a competent cells, to get the recombinant plasmid propagation of MSP1 gene. Blood that was positive for P. falciparum was molecularly processed, starting with genomic DNA isolation and then followed by PCR amplification, ligation into pJET1.2/blunt vector, and transformation into E. coli DH5α using the heat shock transformation method. The process was ended with PCR confirmation to confirm MSP1 gene insertion. The results showed that the presence of the  MSP1 gene in pJET1.2/blunt was successfully confirmed through PCR. From a total of 10 positive colonies grown in liquid culture,  plasmid was isolated. Electropherogram result presented bands  of about 1049bp, indicating the presence of the MSP1 gene in plasmid. Hence, MSP1 gene cloning using pJET1.2/blunt cloning vector and competent cell E. coli DH5α has been successfully performed.  Key words: Heat shock, ligation, MSP-1, P. falciparum, transformation

One particular Anaplasma phagocytophilum ecotype infects cattle in the Camargue, France.

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Dugat, Thibaud; Leblond, Agnès; Keck, Nicolas; Lagrée, Anne-Claire; Desjardins, Isabelle; Joulié, Aurélien; Pradier, Sophie; Durand, Benoit; Boulouis, Henri-Jean; Haddad, Nadia

2017-08-02

Anaplasma phagocytophilum is a zoonotic tick-borne pathogen responsible for granulocytic anaplasmosis, a mild to a severe febrile disease that affects man and several animal species, including cows and horses. In Europe, I. ricinus is the only proven vector for this pathogen, but studies suggest that other tick genera and species could be involved in its transmission. Our objective was to assess the presence and genetic diversity of A. phagocytophilum in domestic animals and different tick species from the Camargue region, located in the south of France. A total of 140 ticks and blood samples from 998 cattle and 337 horses were collected in Camargue and tested for the presence of A. phagocytophilum DNA by msp2 quantitative real-time PCR. Molecular typing with four markers was performed on positive samples. Anaplasma phagocytophilum DNA was detected in 6/993 (0.6%) cows, 1/20 (5%) Haemaphysalis punctata, 1/57 (1.75%) Rhipicephalus pusillus, and was absent in horses (0%). All cattle A. phagocytophilum presented a profile identical to an A. phagocytophilum variant previously detected in Dermacentor marginatus, Hyalomma marginatum, and Rhipicephalus spp. in Camargue. Our results demonstrate that one particular A. phagocytophilum variant infects cattle in Camargue, where I. ricinus is supposed to be rare or even absent. Dermacentor marginatus, Rhipicephalus spp. and Hyalomma spp., and possibly other tick species could be involved in the transmission of this variant in this region.

Effect of milk production system on the enzootic stability to Anaplasma marginale and Babesia bovis in calves in the Campo das Vertentes region of Minas Gerais state, BrazilEfeito do sistema de produção de leite sobre a estabilidade enzoótica para Anaplasma marginale e Babesia bovis em bezerras na região do Campo das Vertentes de Minas Gerais, Brasil

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Christiane Maria Barcellos Magalhães Rocha

2012-04-01

Full Text Available We conducted a cross-sectional observational study, in order to determine the frequency of anti-A. marginale and anti-B. bovis antibodies in calves from four to 12 months of age from ten farms that producing B type milk and an equal number that produce raw milk refrigerated, located in the Campo das Vertentes region of Minas Gerais state, in the period September 2008 to August 2009. Blood smears were performed, serologic testing by indirect immunofluorescence technique (IFAT, given the packed cell volume, rickettsemia, and the clinical scores of animals infected by A. marginale. In the farms that produce B type milk, the overall average frequency of seropositive calves was 94.47% (166/176 and 89.20% (157/176 for A. marginale and B. bovis, respectively. Already on the farms that produce raw milk refrigerated, the overall average frequency of A. marginale was 92.59% (149/161 and for B. bovis from 86.30% (139/161, and there was no significant difference (p > 0.05 in the frequency of calves infected for both hemoparasitic between the two systems of milk production. Statistically significant (p 0.05 among calves from properties that produce B type milk and raw milk refrigerated. The results of this study indicate that, in the Campos das Vertentes region of Minas Gerais, the production system does not interfere with the enzootic stability for A. marginale and B. bovis in calves from dairy farms B milk or raw milk refrigerated, with low probability of anaplasmosis and/or babesiosis in adults animals.Foi realizado um estudo observacional do tipo transversal, com o objetivo de determinar a frequência de anticorpos anti-A. marginale e B. bovis em 337 bezerras com idade entre quatro a 12 meses, oriundas de dez propriedades produtoras de leite B e igual número de fazendas de leite cru refrigerado (leite C, na região do Campo das Vertentes de Minas Gerais, no período de setembro de 2008 a agosto de 2009. Foram realizados esfregaços sangu

Anaplasma phagocytophilum MSP4 and HSP70 Proteins Are Involved in Interactions with Host Cells during Pathogen Infection

Czech Academy of Sciences Publication Activity Database

Contreras, M.; Alberdi, P.; Mateos-Hernández, L.; Fernández de Mera, I.G.; García-Pérez, A. L.; Vancová, Marie; Villar, M.; Ayllón, N.; Cabezas Cruz, Alejandro; Valdés, James J.; Stuen, S.; Gortazar, C.; de la Fuente, J.

2017-01-01

Roč. 7, JUL 5 (2017), č. článku 307. ISSN 2235-2988 EU Projects: European Commission(XE) 278976 - ANTIGONE Institutional support: RVO:60077344 Keywords : anaplasmosis * immunology * hl60 * tick * vaccine * sheep * Anaplasma phagocytophilum Subject RIV: EC - Immunology OBOR OECD: Immunology Impact factor: 4.300, year: 2016

ORF Alignment: NC_004842 [GENIUS II[Archive

Lifescience Database Archive (English)

Full Text Available [Anaplasma ... marginale str. St. Maries] ... Length = 114 ... Query: 16 ... GVNKVILIGHLGKDPDVRVMQNGKEMASFSLATSESWL...DKASGARTEKTEWHSIVVFSEG 75 ... GVNKVILIGHLGKDPDVRVMQNGKEMASFSLATSESWL...DKASGARTEKTEWHSIVVFSEG Sbjct: 1 ... GVNKVILIGHLGKDPDVRVMQNGKEMASFSLATSESWLDKASGARTEKTEWHSIVVFSEG 60 ...

Quantifying the importance of MSP1-19 as a target of growth-inhibitory and protective antibodies against Plasmodium falciparum in humans.

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Danny W Wilson

Full Text Available BACKGROUND: Antibodies targeting blood stage antigens are important in protection against malaria, but the key targets and mechanisms of immunity are not well understood. Merozoite surface protein 1 (MSP1 is an abundant and essential protein. The C-terminal 19 kDa region (MSP1-19 is regarded as a promising vaccine candidate and may also be an important target of immunity. METHODOLOGY/FINDINGS: Growth inhibitory antibodies against asexual-stage parasites and IgG to recombinant MSP1-19 were measured in plasma samples from a longitudinal cohort of 206 children in Papua New Guinea. Differential inhibition by samples of mutant P. falciparum lines that expressed either the P. falciparum or P. chabaudi form of MSP1-19 were used to quantify MSP1-19 specific growth-inhibitory antibodies. The great majority of children had detectable IgG to MSP1-19, and high levels of IgG were significantly associated with a reduced risk of symptomatic P. falciparum malaria during the 6-month follow-up period. However, there was little evidence of PfMSP1-19 specific growth inhibition by plasma samples from children. Similar results were found when testing non-dialysed or dialysed plasma, or purified antibodies, or when measuring growth inhibition in flow cytometry or microscopy-based assays. Rabbit antisera generated by immunization with recombinant MSP1-19 demonstrated strong MSP1-19 specific growth-inhibitory activity, which appeared to be due to much higher antibody levels than human samples; antibody avidity was similar between rabbit antisera and human plasma. CONCLUSIONS/SIGNIFICANCE: These data suggest that MSP1-19 is not a major target of growth inhibitory antibodies and that the protective effects of antibodies to MSP1-19 are not due to growth inhibitory activity, but may instead be mediated by other mechanisms. Alternatively, antibodies to MSP1-19 may act as a marker of protective immunity.

Epidemiology of Babesia, Anaplasma and Trypanosoma species using a new expanded reverse line blot hybridization assay.

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Paoletta, Martina Soledad; López Arias, Ludmila; de la Fournière, Sofía; Guillemi, Eliana Carolina; Luciani, Carlos; Sarmiento, Néstor Fabián; Mosqueda, Juan; Farber, Marisa Diana; Wilkowsky, Silvina Elizabeth

2018-02-01

Vector-borne hemoparasitic infections are a major problem that affects livestock industries worldwide, particularly in tropical and subtropical regions. In this work, a reverse line blot (RLB) hybridization assay was developed for the simultaneous detection and identification of Anaplasma, Babesia and bovine trypanosomes, encompassing in this way the most relevant hemoparasites that affect cattle. A total of 186 bovine blood samples collected from two different ecoepidemiological regions of northeast Argentina, with and without tick control, were analyzed with this new RLB. High diversity of parasites, such as Babesia bovis, B. bigemina, Anaplasma marginale and three different Trypanosoma species, was found. High rates of coinfections were also detected, and significant differences were observed not only in the prevalence of parasites but also in the level of coinfections between the two analyzed areas. Regarding the Trypanosoma genus, we provide molecular evidence of the presence of T. vivax and T. theileri for the first time in Argentina. Besides, since the RLB is a prospective tool, it allowed the identification of a yet unknown bovine trypanosome which could not be assigned to any of the bovine species known so far. In the present study we provide new insights on the prevalence of several pathogens that directly impact on livestock production in Argentina. The RLB assay developed here allows to identify simultaneously numerous pathogenic species which can also be easily expanded to detect other blood borne pathogens. These characteristics make the RLB hybridization assay an essential tool for epidemiological survey of all vector-borne pathogens. Copyright © 2017 Elsevier GmbH. All rights reserved.

Natural Anaplasma phagocytophilum infection in ticks from a forest area of Selenge province, Mongolia

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G Javkhlan

2014-03-01

Full Text Available Anaplasma phagocytophilum is a zoonotic agent of public health importance, infecting both humans and animals. An investigation of the presence of Anaplasma phagocytophilum as well as Anaplasma platys was conducted in a forest area of Selenge province, Mongolia, where ticks are widely distributed and tick-borne diseases are highly endemic. Ticks were collected and tested using polymerase chain reaction based on groEL methodology. Anaplasma phagocytophilum was detected in 14 (6% of Ixodes persulcatus ticks and four (1% Dermacentor nuttalli ticks; infection of Anaplasma platys was detected in 1% of Ixodes persulcatus ticks and 10% of Dermacentor nuttalli ticks. The phylogenetic tree showed that the Anaplasma phagocytophilum clustered with the Russian group, most likely due to similar geographical locations. This finding is significant for both veterinary and public health officials given that these agents can cause both animal and human illness.

Formation of the food vacuole in Plasmodium falciparum: a potential role for the 19 kDa fragment of merozoite surface protein 1 (MSP1(19.

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Anton R Dluzewski

2008-08-01

Full Text Available Plasmodium falciparum Merozoite Surface Protein 1 (MSP1 is synthesized during schizogony as a 195-kDa precursor that is processed into four fragments on the parasite surface. Following a second proteolytic cleavage during merozoite invasion of the red blood cell, most of the protein is shed from the surface except for the C-terminal 19-kDa fragment (MSP1(19, which is still attached to the merozoite via its GPI-anchor. We have examined the fate of MSP1(19 during the parasite's subsequent intracellular development using immunochemical analysis of metabolically labeled MSP1(19, fluorescence imaging, and immuno-electronmicroscopy. Our data show that MSP1(19 remains intact and persists to the end of the intracellular cycle. This protein is the first marker for the biogenesis of the food vacuole; it is rapidly endocytosed into small vacuoles in the ring stage, which coalesce to form the single food vacuole containing hemozoin, and persists into the discarded residual body. The food vacuole is marked by the presence of both MSP1(19 and the chloroquine resistance transporter (CRT as components of the vacuolar membrane. Newly synthesized MSP1 is excluded from the vacuole. This behavior indicates that MSP1(19 does not simply follow a classical lysosome-like clearance pathway, instead, it may play a significant role in the biogenesis and function of the food vacuole throughout the intra-erythrocytic phase.

Solution structure of a Plasmodium falciparum AMA-1/MSP 1 chimeric protein vaccine candidate (PfCP-2.9 for malaria

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Jin Changwen

2010-03-01

Full Text Available Abstract Background The Plasmodium falciparum chimeric protein PfCP-2.9 is a promising asexual-stage malaria vaccine evaluated in clinical trials. This chimeric protein consists of two cysteine-rich domains: domain III of the apical membrane antigen 1 (AMA-1 [III] and the C-terminal region of the merozoite surface protein 1 (MSP1-19. It has been reported that the fusion of these two antigens enhanced their immunogenicity and antibody-mediated inhibition of parasite growth in vitro. Methods The 15N-labeled and 13C/15N-labeled PfCP-2.9 was produced in Pichia pastoris for nuclear magnetic resonance (NMR structure analysis. The chemical shift assignments of PfCP-2.9 were compared with those previously reported for the individual domains (i.e., PfAMA-1(III or PfMSP 1-19. The two-dimensional spectra and transverse relaxation rates (R2 of the PfMSP1-19 alone were compared with that of the PfCP-2.9. Results Confident backbone assignments were obtained for 122 out of 241 residues of PfCP-2.9. The assigned residues in PfCP-2.9 were very similar to those previously reported for the individual domains. The conformation of the PfMSP1-19 in different constructs is essentially the same. Comparison of transverse relaxation rates (R2 strongly suggests no weak interaction between the domains. Conclusions These data indicate that the fusion of AMA-1(III and MSP1-19 as chimeric protein did not change their structures, supporting the use of the chimeric protein as a potential malaria vaccine.

Lineage-specific positive selection at the merozoite surface protein 1 (msp1 locus of Plasmodium vivax and related simian malaria parasites

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Kawai Satoru

2010-02-01

Full Text Available Abstract Background The 200 kDa merozoite surface protein 1 (MSP-1 of malaria parasites, a strong vaccine candidate, plays a key role during erythrocyte invasion and is a target of host protective immune response. Plasmodium vivax, the most widespread human malaria parasite, is closely related to parasites that infect Asian Old World monkeys, and has been considered to have become a parasite of man by host switch from a macaque malaria parasite. Several Asian monkey parasites have a range of natural hosts. The same parasite species shows different disease manifestations among host species. This suggests that host immune responses to P. vivax-related malaria parasites greatly differ among host species (albeit other factors. It is thus tempting to invoke that a major immune target parasite protein such as MSP-1 underwent unique evolution, depending on parasite species that exhibit difference in host range and host specificity. Results We performed comparative phylogenetic and population genetic analyses of the gene encoding MSP-1 (msp1 from P. vivax and nine P. vivax-related simian malaria parasites. The inferred phylogenetic tree of msp1 significantly differed from that of the mitochondrial genome, with a striking displacement of P. vivax from a position close to P. cynomolgi in the mitochondrial genome tree to an outlier of Asian monkey parasites. Importantly, positive selection was inferred for two ancestral branches, one leading to P. inui and P. hylobati and the other leading to P. vivax, P. fieldi and P. cynomolgi. This ancestral positive selection was estimated to have occurred three to six million years ago, coinciding with the period of radiation of Asian macaques. Comparisons of msp1 polymorphisms between P. vivax, P. inui and P. cynomolgi revealed that while some positively selected amino acid sites or regions are shared by these parasites, amino acid changes greatly differ, suggesting that diversifying selection is acting species

The risk of developing cervical cancer in Mexican women is associated to CYP1A1 MspI polymorphism.

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Juárez-Cedillo, Teresa; Vallejo, Maite; Fragoso, José Manuel; Hernández-Hernández, Dulce Maria; Rodríguez-Pérez, José Manuel; Sánchez-García, Sergio; del Carmen García-Peña, María; García-Carrancá, Alejandro; Mohar-Betancourt, Alejandro; Granados, Julio; Vargas-Alarcón, Gilberto

2007-07-01

The aim of the study was to evaluate the association of two CYP1A1 polymorphisms (Msp1 and exon 7) with cervical cancer in Mexican women considering their smoking habit. The polymorphisms were determined in 310 individuals (155 with cervical cancer and 155 healthy controls). Women with MspI T/C or C/C showed increased risk of developing cervical cancer (3.7- and 8.3-fold increase, respectively) compared to women with T/T genotype. When smoking habit was considered, the risk for non-smokers with T/C and C/C genotypes was similar (5.2 and 4.1, respectively), whereas smoking women with C/C genotype showed a 19.4-fold increase of cervical cancer. Number of child births, number of sexual partners and marital status were strong risk factors for developing cervical cancer in women with T/T genotype; however, in women with T/C genotype, only the number of child births and sexual partners had a significant influence. These results suggest an important role of the CYP1A1 MspI polymorphism in the risk of developing cervical cancer.

Phase 1 trial of the Plasmodium falciparum blood stage vaccine MSP1(42-C1/Alhydrogel with and without CPG 7909 in malaria naïve adults.

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Ruth D Ellis

2010-01-01

Full Text Available Merozoite surface protein 1(42 (MSP1(42 is a leading blood stage malaria vaccine candidate. In order to induce immune responses that cover the major antigenic polymorphisms, FVO and 3D7 recombinant proteins of MSP1(42 were mixed (MSP1(42-C1. To improve the level of antibody response, MSP1(42-C1 was formulated with Alhydrogel plus the novel adjuvant CPG 7909.A Phase 1 clinical trial was conducted in healthy malaria-naïve adults at the Center for Immunization Research in Washington, D.C., to evaluate the safety and immunogenicity of MSP1(42-C1/Alhydrogel +/- CPG 7909. Sixty volunteers were enrolled in dose escalating cohorts and randomized to receive three vaccinations of either 40 or 160 microg protein adsorbed to Alhydrogel +/- 560 microg CPG 7909 at 0, 1 and 2 months.Vaccinations were well tolerated, with only one related adverse event graded as severe (Grade 3 injection site erythema and all other vaccine related adverse events graded as either mild or moderate. Local adverse events were more frequent and severe in the groups receiving CPG. The addition of CPG enhanced anti-MSP1(42 antibody responses following vaccination by up to 49-fold two weeks after second immunization and 8-fold two weeks after the third immunization when compared to MSP1(42-C1/Alhydrogel alone (p<0.0001. After the third immunization, functionality of the antibody was tested by an in vitro growth inhibition assay. Inhibition was a function of antibody titer, with an average of 3% (range -2 to 10% in the non CPG groups versus 14% (3 to 32% in the CPG groups.The favorable safety profile and high antibody responses induced with MSP1(42-C1/Alhydrogel + CPG 7909 are encouraging. MSP1(42-C1/Alhydrogel is being combined with other blood stage antigens and will be taken forward in a formulation adjuvanted with CPG 7909.ClinicalTrials.gov Identifier: NCT00320658.

CYP1A1 MspI Polymorphism and Cervical Carcinoma Risk in the Multi-Ethnic Population of Malaysia: a Case-Control Study.

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Tan, Yee Hock; Sidik, Shiran Mohd; Syed Husain, Sharifah Noor Akmal; Lye, Munn Sann; Chong, Pei Pei

2016-01-01

Tobacco smoking is considered a risk factor for cervical cancer development due to the presence of tobacco based carcinogenic metabolites in cervical cells of female smokers. In this study, we investigated the role of the T3801C (MspI) polymorphism of CYP1A1, a gene encoding an enzyme necessary for the initiation of tobacco based carcinogen metabolism, on cervical cancer risk. The T to C substitution may alter CYP1A1 activities, potentially elevating cervical cancer risk. Since results of gene-disease association studies vary according to the study population, the multi-ethnic population of Malaysia provides an excellent representative cohort for identifying and comparing the cervical cancer risk among the 3 major ethnics in Southeast Asia in relation to CYP1A1 MspI polymorphism. A total of 195 Thin Prep Pap smear samples from HPV negative and cancer free females were randomly selected as controls while 106 formalin fixed paraffin embedded samples from females with invasive cervical cancer were randomly selected for the cases group. The polymorphisms were identified using restriction fragment length polymorphism (RFLP) PCR. We found no significant associations between CYP1A1 MspI polymorphism and cervical cancer in the general Malaysian female population. However, upon ethnic stratification, the variant C/C genotype was significantly associated with a 4.66-fold increase in cervical cancer risk in Malay females (95% CI= 1.21-17.9; p=0.03). No significant association was observed in the Chinese and Indian females. Additionally, there were no significant associations in the dominant model and allele frequency model analysis in both the general and ethnically stratified female population of Malaysia. Our findings suggest that the C/C genotype of CYP1A1 MspI polymorphism is associated with the development of cervical carcinoma in the Malay females of Malaysia.

Global phylogeography of the avian malaria pathogen Plasmodium relictum based on MSP1 allelic diversity

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Hellgren, Olof; Atkinson, Carter T.; Bensch, Staffan; Albayrak, Tamer; Dimitrov, Dimitar; Ewen, John G.; Kim, Kyeong Soon; Lima, Marcos R.; Martin, Lynn; Palinauskas, Vaidas; Ricklefs, Robert; Sehgal, Ravinder N. M.; Gediminas, Valkiunas; Tsuda, Yoshio; Marzal, Alfonso

2015-01-01

Knowing the genetic variation that occurs in pathogen populations and how it is distributed across geographical areas is essential to understand parasite epidemiology, local patterns of virulence, and evolution of host-resistance. In addition, it is important to identify populations of pathogens that are evolutionarily independent and thus ‘free’ to adapt to hosts and environments. Here, we investigated genetic variation in the globally distributed, highly invasive avian malaria parasite Plasmodium relictum, which has several distinctive mitochondrial haplotyps (cyt b lineages, SGS1, GRW11 and GRW4). The phylogeography of P. relictum was accessed using the highly variable nuclear gene merozoite surface protein 1 (MSP1), a gene linked to the invasion biology of the parasite. We show that the lineage GRW4 is evolutionarily independent of GRW11 and SGS1 whereas GRW11 and SGS1 share MSP1 alleles and thus suggesting the presence of two distinct species (GRW4 versus SGS1 and GRW11). Further, there were significant differences in the global distribution of MSP1 alleles with differences between GRW4 alleles in the New and the Old World. For SGS1, a lineage formerly believed to have both tropical and temperate transmission, there were clear differences in MSP1 alleles transmitted in tropical Africa compared to the temperate regions of Europe and Asia. Further, we highlight the occurrence of multiple MSP1 alleles in GRW4 isolates from the Hawaiian Islands, where the parasite has contributed to declines and extinctions of endemic forest birds since it was introduced. This study stresses the importance of multiple independent loci for understanding patterns of transmission and evolutionary independence across avian malaria parasites.

Milk fat globule-epidermal growth factor-factor VIII-derived peptide MSP68 is a cytoskeletal immunomodulator of neutrophils that inhibits Rac1.

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Hendricks, Louie; Aziz, Monowar; Yang, Weng-Lang; Nicastro, Jeffrey; Coppa, Gene F; Symons, Marc; Wang, Ping

2017-02-01

Prolonged neutrophil infiltration leads to exaggerated inflammation and tissue damage during sepsis. Neutrophil migration requires rearrangement of their cytoskeleton. Milk fat globule-epidermal growth factor-factor VIII-derived short peptide 68 (MSP68) has recently been shown to be beneficial in sepsis-induced tissue injury and mortality. We hypothesize that MSP68 inhibits neutrophil migration by modulating small GTPase Rac1-dependent cytoskeletal rearrangements. Bone marrow-derived neutrophils (BMDNs) or whole lung digest isolated neutrophils were isolated from 8 to 10 wk old C57BL/6 mice by Percoll density gradient centrifugation. The purity of BMDN was verified by flow cytometry with CD11b/Gr-1 staining. Neutrophils were stimulated with N-formylmethionine-leucine-phenylalanine (f-MLP) (10 nM) in the presence or absence of MSP68 at 10 nM or cecal ligation and puncture (CLP) was used to induce sepsis, and MSP68 was administered at 1 mg/kg intravenously. Cytoskeletal organization was assessed by phalloidin staining, followed by analysis using fluorescence microscopy. Activity of the Rac1 GTPase in f-MLP or CLP-activated BMDN in the presence or absence of MSP68 was assessed by GTPase enzyme-linked immunosorbent assay. Mitogen-activated protein (MAP) kinase activity was determined by western blot densitometry. BMDN treatment with f-MLP increased cytoskeletal remodeling as revealed by the localization of filamentous actin to the periphery of the neutrophil. By contrast, cells pretreated with MSP68 had considerably reduced filamentous actin polymerization. Cytoskeletal spreading is associated with the activation of the small GTPase Rac1. We found BMDN-treated with f-MLP or that were exposed to sepsis by CLP had increased Rac1 signaling, whereas the cells pretreated with MSP68 had significantly reduced Rac1 activation (P Rac1-MAP kinase-mediated neutrophil motility. Thus, MSP68 is a novel therapeutic candidate for regulating inflammation and tissue damage caused

Nanoscopic surfactant behavior of the porin MspA in aqueous media

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Ayomi S. Perera

2013-04-01

Full Text Available The mycobacterial porin MspA is one of the most stable channel proteins known to date. MspA forms vesicles at low concentrations in aqueous buffers. Evidence from dynamic light scattering, transmission electron microscopy and zeta-potential measurements by electrophoretic light scattering indicate that MspA behaves like a nanoscale surfactant. The extreme thermostability of MspA allows these investigations to be carried out at temperatures as high as 343 K, at which most other proteins would quickly denature. The principles of vesicle formation of MspA as a function of temperature and the underlying thermodynamic factors are discussed here. The results obtained provide crucial evidence in support of the hypothesis that, during vesicle formation, nanoscopic surfactant molecules, such as MspA, deviate from the principles underlined in classical surface chemistry.

MspI and PvuII polymorphisms in the Na,K-ATPase. beta. subunit gene ATP1B1

Energy Technology Data Exchange (ETDEWEB)

Shull, M.M.; Pugh, D.G.; Lane, L.K.; Lingrel, J.B. (Univ. of Cincinnati College of Medicine, OH (USA))

1990-02-25

ATP1B HH1.2 is a 1.2 kb HindIII fragment from the 3{prime} portion of the human Na,K-ATPase {beta} subunit gene, ATP1B1. MspI identifies a two allele polymorphism (M1: 6.7 kb, M2: 5.3 kb). PvuII also detects a two-allele polymorphism (P1: 5.1 kb, P2: 4.7 kb). ATP1B1 has been assigned to chromosome 1q by somatic cell hybrid analysis. Codominant segregation of the MspI RFLP was observed in one two-generation family (5 individuals). Codominant segregation of the PvuII RFLP was observed in a two-generation (8 individuals) and a three-generation (12 individuals) family.

A molecular epidemiological survey of Babesia, Hepatozoon, Ehrlichia and Anaplasma infections of dogs in Japan

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KUBO, Shotaro; TATENO, Morihiro; ICHIKAWA, Yasuaki; ENDO, Yasuyuki

2015-01-01

Tick-borne diseases are often encountered in canine clinical practice. In the present study, a molecular epidemiological survey of dogs in Japan was conducted to understand the prevalence and geographical distribution of Babesia spp., Hepatozoon spp., Ehrlichia spp. and Anaplasma spp. Pathogen-derived DNA in blood samples obtained from 722 dogs with a history of exposure to ticks and/or fleas was examined by PCR. The prevalence of Babesia gibsoni, Babesia odocoilei-like species, Hepatozoon canis and Ehrlichia spp./Anaplasma spp. was 2.4% (16/722), 0.1% (1/722), 2.5% (18/722) and 1.5% (11/722), respectively. While B. gibsoni and Ehrlichia spp./Anaplasma spp. were detected in the western part of Japan, H. canis was detected in Tohoku area in addition to western and central parts of Japan. PMID:25947226

An O-Methyltransferase Is Required for Infection of Tick Cells by Anaplasma phagocytophilum.

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Oliva Chávez, Adela S; Fairman, James W; Felsheim, Roderick F; Nelson, Curtis M; Herron, Michael J; Higgins, LeeAnn; Burkhardt, Nicole Y; Oliver, Jonathan D; Markowski, Todd W; Kurtti, Timothy J; Edwards, Thomas E; Munderloh, Ulrike G

2015-01-01

Anaplasma phagocytophilum, the causative agent of Human Granulocytic Anaplasmosis (HGA), is an obligately intracellular α-proteobacterium that is transmitted by Ixodes spp ticks. However, the pathogen is not transovarially transmitted between tick generations and therefore needs to survive in both a mammalian host and the arthropod vector to complete its life cycle. To adapt to different environments, pathogens rely on differential gene expression as well as the modification of proteins and other molecules. Random transposon mutagenesis of A. phagocytophilum resulted in an insertion within the coding region of an o-methyltransferase (omt) family 3 gene. In wild-type bacteria, expression of omt was up-regulated during binding to tick cells (ISE6) at 2 hr post-inoculation, but nearly absent by 4 hr p.i. Gene disruption reduced bacterial binding to ISE6 cells, and the mutant bacteria that were able to enter the cells were arrested in their replication and development. Analyses of the proteomes of wild-type versus mutant bacteria during binding to ISE6 cells identified Major Surface Protein 4 (Msp4), but also hypothetical protein APH_0406, as the most differentially methylated. Importantly, two glutamic acid residues (the targets of the OMT) were methyl-modified in wild-type Msp4, whereas a single asparagine (not a target of the OMT) was methylated in APH_0406. In vitro methylation assays demonstrated that recombinant OMT specifically methylated Msp4. Towards a greater understanding of the overall structure and catalytic activity of the OMT, we solved the apo (PDB_ID:4OA8), the S-adenosine homocystein-bound (PDB_ID:4OA5), the SAH-Mn2+ bound (PDB_ID:4PCA), and SAM- Mn2+ bound (PDB_ID:4PCL) X-ray crystal structures of the enzyme. Here, we characterized a mutation in A. phagocytophilum that affected the ability of the bacteria to productively infect cells from its natural vector. Nevertheless, due to the lack of complementation, we cannot rule out secondary mutations.

An O-Methyltransferase Is Required for Infection of Tick Cells by Anaplasma phagocytophilum.

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Adela S Oliva Chávez

Full Text Available Anaplasma phagocytophilum, the causative agent of Human Granulocytic Anaplasmosis (HGA, is an obligately intracellular α-proteobacterium that is transmitted by Ixodes spp ticks. However, the pathogen is not transovarially transmitted between tick generations and therefore needs to survive in both a mammalian host and the arthropod vector to complete its life cycle. To adapt to different environments, pathogens rely on differential gene expression as well as the modification of proteins and other molecules. Random transposon mutagenesis of A. phagocytophilum resulted in an insertion within the coding region of an o-methyltransferase (omt family 3 gene. In wild-type bacteria, expression of omt was up-regulated during binding to tick cells (ISE6 at 2 hr post-inoculation, but nearly absent by 4 hr p.i. Gene disruption reduced bacterial binding to ISE6 cells, and the mutant bacteria that were able to enter the cells were arrested in their replication and development. Analyses of the proteomes of wild-type versus mutant bacteria during binding to ISE6 cells identified Major Surface Protein 4 (Msp4, but also hypothetical protein APH_0406, as the most differentially methylated. Importantly, two glutamic acid residues (the targets of the OMT were methyl-modified in wild-type Msp4, whereas a single asparagine (not a target of the OMT was methylated in APH_0406. In vitro methylation assays demonstrated that recombinant OMT specifically methylated Msp4. Towards a greater understanding of the overall structure and catalytic activity of the OMT, we solved the apo (PDB_ID:4OA8, the S-adenosine homocystein-bound (PDB_ID:4OA5, the SAH-Mn2+ bound (PDB_ID:4PCA, and SAM- Mn2+ bound (PDB_ID:4PCL X-ray crystal structures of the enzyme. Here, we characterized a mutation in A. phagocytophilum that affected the ability of the bacteria to productively infect cells from its natural vector. Nevertheless, due to the lack of complementation, we cannot rule out secondary

ChAd63-MVA-vectored blood-stage malaria vaccines targeting MSP1 and AMA1: assessment of efficacy against mosquito bite challenge in humans.

Science.gov (United States)

Sheehy, Susanne H; Duncan, Christopher J A; Elias, Sean C; Choudhary, Prateek; Biswas, Sumi; Halstead, Fenella D; Collins, Katharine A; Edwards, Nick J; Douglas, Alexander D; Anagnostou, Nicholas A; Ewer, Katie J; Havelock, Tom; Mahungu, Tabitha; Bliss, Carly M; Miura, Kazutoyo; Poulton, Ian D; Lillie, Patrick J; Antrobus, Richard D; Berrie, Eleanor; Moyle, Sarah; Gantlett, Katherine; Colloca, Stefano; Cortese, Riccardo; Long, Carole A; Sinden, Robert E; Gilbert, Sarah C; Lawrie, Alison M; Doherty, Tom; Faust, Saul N; Nicosia, Alfredo; Hill, Adrian V S; Draper, Simon J

2012-12-01

The induction of cellular immunity, in conjunction with antibodies, may be essential for vaccines to protect against blood-stage infection with the human malaria parasite Plasmodium falciparum. We have shown that prime-boost delivery of P. falciparum blood-stage antigens by chimpanzee adenovirus 63 (ChAd63) followed by the attenuated orthopoxvirus MVA is safe and immunogenic in healthy adults. Here, we report on vaccine efficacy against controlled human malaria infection delivered by mosquito bites. The blood-stage malaria vaccines were administered alone, or together (MSP1+AMA1), or with a pre-erythrocytic malaria vaccine candidate (MSP1+ME-TRAP). In this first human use of coadministered ChAd63-MVA regimes, we demonstrate immune interference whereby responses against merozoite surface protein 1 (MSP1) are dominant over apical membrane antigen 1 (AMA1) and ME-TRAP. We also show that induction of strong cellular immunity against MSP1 and AMA1 is safe, but does not impact on parasite growth rates in the blood. In a subset of vaccinated volunteers, a delay in time to diagnosis was observed and sterilizing protection was observed in one volunteer coimmunized with MSP1+AMA1-results consistent with vaccine-induced pre-erythrocytic, rather than blood-stage, immunity. These data call into question the utility of T cell-inducing blood-stage malaria vaccines and suggest that the focus should remain on high-titer antibody induction against susceptible antigen targets.

Genotipificación de Plasmodium falciparum por PCR múltiple por medio de los genes msp1, msp2 y glurp, en cuatro localidades de Colombia

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Sandra Milena Barrera

2010-12-01

Conclusiones. Nuestros resultados demuestran que el marcador molecular msp1 no proporciona información útil para diferenciar las poblaciones parasitarias de P. falciparum. El gen msp2 es apropiado para evaluar la diversidad genética, pero requiere ensayos más finos que permitan diferenciar claramente el polimorfismo de tamaño en las dos familias. Los resultados obtenidos con el gen glurp evidenciaron una gran diversidad genética en las poblaciones de P. falciparum que circulan en el país y sugieren que este gen puede ser útil para diferenciar nuevas infecciones de infecciones recurrentes o recrudecimientos.

Anaplasma, Ehrlichia and Rickettsia species infections in cats

NARCIS (Netherlands)

Pennisi, Maria Grazia; Hofmann-Lehmann, Regina; Radford, Alan D; Tasker, Séverine; Belák, Sándor; Addie, Diane D; Boucraut-Baralon, Corine; Egberink, Herman; Frymus, Tadeusz; Gruffydd-Jones, Tim; Hartmann, Katrin; Horzinek, Marian C; Hosie, Margaret J; Lloret, Albert; Lutz, Hans; Marsilio, Fulvio; Thiry, Etienne; Truyen, Uwe; Möstl, Karin

2017-01-01

OVERVIEW: Anaplasma species, Ehrlichia species and Rickettsia species are vector-borne pathogens infecting a wide variety of mammals, but causing disease in very few of them. Infection in cats: Anaplasma phagocytophilum is the most important feline pathogen among these rickettsial organisms, and

Molecular and serological prevalence of Toxoplasma gondii and Anaplasma spp. infection in goats from Chongqing Municipality, China

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Zhou Zuoyong

2018-01-01

Full Text Available Toxoplasmosis and anaplasmosis are severe zoonotic diseases, the former caused by Toxoplasma gondii and the latter by Anaplasma spp. In the present study, 332 goat blood samples were randomly collected from Chongqing Municipality, China to screen for T. gondii and Anaplasma spp. We used a polymerase chain reaction (PCR to detect DNA, and enzyme-linked immunosorbent assay (ELISA to test for T. gondii antibodies. The prevalence of T. gondii and Anaplasma spp. was 38% and 35% respectively by PCR, and 42% for T. gondii antibodies by ELISA. The co-infection rate by T. gondii and Anaplasma was 13%, where the two predominant pathogens co-infecting were Anaplasma phagocytophilum + A. bovis (10%, followed by T. gondii + A. phagocytophilum (9.64%. While co-infection by three pathogens varied ranging from 1.81% to 5.72%, less than 1% of goats were found to be positive for four pathogens. This is the first investigation of T. gondii and Anaplasma spp. infection in goats from Chongqing.

MspI and PvuII polymorphisms in the Na,K-ATPase. alpha. subunit related gene ATP1AL1

Energy Technology Data Exchange (ETDEWEB)

Shull, M.M.; Pugh, D.G.; Lingrel, J.B. (Univ. of Cincinnati, OH (USA))

1990-01-11

ATP1AL1 78-1-3 is a 0.56 kb genomic EcoRI-XbaI fragment from within the Na,K-ATPase {alpha} subunit related gene, previously referred to as {alpha}D on chromosome 13. The fragment was subcloned into pIBI31. MspI identifies a two-allele polymorphism (M1: 2.8 kb, M2: 2.5 kb). PvuII, which cuts within the probe sequence, detects two two-allele polymorphism (A1: 6.0 kb, A2: 5.7 kb, B1: 1.3 kb, B2: 1.1 kb). A1 and A2 appear to result from an insertion/deletion polymorphism that is also identified by MspI. ATP1AL1 78-1-3 has been assigned to chromosome 13q by somatic cell hybrid analysis. Codominant segregation of the RELPs was observed in 2 two-generation families.

Common MspI RFLP of the human fibronectin gene (FN1)

Energy Technology Data Exchange (ETDEWEB)

Gardella, R; Colombi, M; Barlati, S

1988-02-25

The probe used, pFH154, contains a sequence of 2.5 kb of the fibronectin cDNA inserted in pAT153/Pvu/8 plasmid vector. The recombinant plasmid and the HindIII/BamHI purified cDNA insert give identical patterns. MspI (C/CGG) (Promega Biotec) detects invariant bands at 8.5, 4.2, 2.5, 2.1, 1.55, 1.4, 1.1 and 0.75 kb in addition to a simple two allele polymorphism with a band at either 0.35 kb (F1) or 2.7 kb (F2). Co-dominant segregation was demonstrated in 6 families.

Intrauterine Transmission of Anaplasma phagocytophilum in Persistently Infected Lambs

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Snorre Stuen

2018-02-01

Full Text Available Anaplasma phagocytophilum, which causes the disease tick-borne fever (TBF, is the most important tick-borne pathogen in European animals. TBF may contribute to severe welfare challenges and economic losses in the Norwegian sheep industry. The bacterium causes a persistent infection in sheep and several other animal species. The objective of this study was to investigate whether intrauterine transmission occurs in persistently infected sheep. The study included thirteen 5–6-month-old unmated ewes, of which twelve were experimentally infected with A. phagocytophilum (GenBank acc. no. M73220. Four to six weeks later, all ewes were mated, and nine became pregnant. Blood samples were collected from these ewes and their offspring. If the lamb died, tissue samples were collected. The samples were analyzed with real-time PCR (qPCR targeting the msp2 gene. PCR-positive samples were further analyzed by semi-nested PCR and 16S rDNA sequencing. A total of 20 lambs were born, of which six died within two days. Six newborn lambs (30% were PCR-positive (qPCR, of which one was verified by 16S rDNA sequencing. The present study indicates that intrauterine transmission of A. phagocytophilum in persistently infected sheep may occur. The importance of these findings for the epidemiology of A. phagocytophilum needs to be further investigated.

Prevalence of Anaplasma phagocytophilum in North Carolina Eastern Black Bears ( Ursus americanus ).

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Westmoreland, Lori S H; Stoskopf, Michael K; Maggi, Ricardo G

2016-10-01

We detected Anaplasma phagocytophilum by DNA amplification in whole blood from free-ranging, hunter-killed American black bears ( Ursus americanus ) from the east coast of North Carolina, US. Molecular prevalence for Anaplasma phagocytophilum was 3% from 68 black bears. No DNA of other Anaplasma or Ehrlichia spp. was identified.

First case of Anaplasma platys infection in a dog from Croatia

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Dyachenko Viktor

2012-03-01

Full Text Available Abstract Background It is known that Anaplasma (A. platys, the causative agent of infectious canine cyclic thrombocytopenia, is endemic in countries of the Mediterranean basin. However, few reports are available from the Balkans. This case report describes a dog, which was imported from Croatia to Germany in May 2010. One month later the dog was presented to a local veterinarian in Germany due to intermittent/recurrent diarrhoea. Diagnostic tests were performed to identify infections caused by Anaplasma spp., Ehrlichia spp., Hepatozoon canis, Babesia spp., Leishmania spp., Borrelia burgdorferi and/or Dirofilaria immitis. Findings Haematological examination of a blood smear revealed basophilic inclusions in thrombocytes, which were confirmed as A. platys with a species-specific real-time PCR. Additionally, an infection with Babesia (B. vogeli was also detected (PCR and serology. No specific antibodies against Anaplasma antigen were detectable. Although the dog showed no specific clinical signs, thrombocytopenia, anaemia and elevated C-reactive protein (CRP were observed. Sequencing of a 1,348-bp partial ribosomal RNA gene revealed highest homology to A. platys sequences from Thailand, Japan and France. Conclusions A. platys was detected for first time in a dog imported from Croatia. As the dog was also co-infected by B. vogeli, unique serological and haematological findings were recorded. Thrombocytopenia, anaemia and elevated values of C-reactive protein were the laboratory test abnormalities observed in this case. A. platys infections should be considered in dogs coming from Croatia and adjacent regions.

Infecção natural por hemoparasitos em bezerros submetidos à quimio-profilaxia aos 30 dias de idade Natural infection by hemoparasites in calves submitted to chemo -prophylaxis wonder 30 days of age

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Rosângela A. Da Silva

2007-09-01

Full Text Available O complexo Tristeza Parasitária acarreta grandes prejuízos à pecuária bovina nacional. Os principais agentes etiológicos são Babesia bigemina, B. bovis e Anaplasma marginale, sendo o carrapato Boophilus microplus o principal vetor. Este trabalho relata a ocorrência de infecção natural por hemoparasitos da tristeza parasitária bovina em 36 bezerros com alta infestação natural por carrapatos e submetidos à quimioprofilaxia aos 30 dias de idade. Babesia bigemina (33,3%, B. bovis (11,1% e A. marginale (13,9% foram detectados em esfregaços sangüíneos de 16 animais (44,4% de diferentes idades. Seis bezerros apresentaram sintomas (16,7% e um morreu (2,8%. O número de casos clínicos foi decorrente de uma associação de fatores, destacando-se a alta infestação precoce por carrapatos e a baixa imunidade passiva em período em que os bezerros ainda não haviam desenvolvido imunidade ativa suficiente.The tick-borne disease (TBD brings great damages to cattle breeding. The most important etiologic agents are Babesia bigemina, B. bovis and Anaplasma marginale, being the tick Boophilus microplus the main vector. This work reports the occurrence of natural infection by hemoparasites of TBD in 36 calves with high ticks natural infestation submitted to chemoprophylaxis with 30 days year-old. The blood smears from animals of different ages were analized and were found B. bigemina (33.3%, B. bovis (11.1% and A. marginale (13.9%. Six animals had clinical symptoms (16.7% and one dead (2.8%. The number of clinical cases ocurred in consequence of an association of factors as high infestation of ticks and low passive immunity in period that calves had not developed enough active immunity.

Immunisation of smallholder dairy cattle against anaplasmosis and babesiosis in Malawi

DEFF Research Database (Denmark)

Tjørnehøj, Kirsten; Lawrence, J. A.; Kafuwa, P. T.

1997-01-01

A field study was conducted in the Southern Region of Malawi to evaluate the possible benefits of immunisation of improved dairy cattle against Anaplasma marginale, Babesia bigemina and Babesia bovis. Friesian crossbred heifers were immunised when they were being reared on Government farms. They ...

Development of a Pulse Oximeter Using MSP430 Microcontroller

OpenAIRE

Gayathri.R; Hepsiba.D

2013-01-01

In this paper,we propose and demonstrate the design of a Pulse Oximeter using MSP430FG439 Microcontroller.The Oxygen Saturation and Heart Rate are the two important key parameters for health monitoring of a patients.The proposed system consists of spo₂ Sensor,MSP430FG439 Pulse Oximeter board With LCD Display,PC.The oxygen saturation is calculated by measuring the ratio between the intensities of two different lights.The heart rate is calculated by measuring the elapsed time between the peaks ...

EL MISTICISMO de los espíritus marginales

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JOSÉ JORGE DE CARVALHO

2001-01-01

Full Text Available EL ARTÍCULO OFRECE UNA VISIÓN PANORÁMICA DE LAS MANIFESTACIONES RELIGIOSAS brasileñas marginales con relación al cristianismo dominante, entre las cuales se destacan el espiritismo kardecista y las varias formas de religiones de origen africano, todas de tradición oral. Un punto común a esas expresiones religiosas es la experiencia del trance y la posesión, que busco teorizar con la idea de la apófasis radical. En la segunda mitad del ensayo presento una antología de textos sagrados afrobrasileños, los cuales comento en una perspectiva de simbología comparada. Ellos señalan la presencia de un misticismo análogo al de los místicos de las llamadas "grandes religiones" o "religiones del libro". Propongo, entonces, incluir esas tradiciones marginales como parte constitutiva de la experiencia religiosa de la humanidad como un todo.

Detection of Rickettsia and Anaplasma from hard ticks in Thailand.

Science.gov (United States)

Malaisri, Premnika; Hirunkanokpun, Supanee; Baimai, Visut; Trinachartvanit, Wachareeporn; Ahantarig, Arunee

2015-12-01

We collected a total of 169 adult hard ticks and 120 nymphs from under the leaves of plants located along tourist nature trails in ten localities. The results present data examining the vector competence of ticks of different genera and the presence of Rickettsia and Anaplasma species. The ticks belonged to three genera, Amblyomma, Dermacentor, and Haemaphysalis, comprising 11 species. Rickettsia bacteria were detected at three collection sites, while Anaplasma bacteria were detected at only one site. Phylogenetic analysis revealed new rickettsia genotypes from Thailand that were closely related to Rickettsia tamurae, Rickettsia monacensis, and Rickettsia montana. This study was also the first to show that Anaplasma bacteria are found in Haemaphysalis shimoga ticks and are closely related evolutionarily to Anaplasma bovis. These results provide additional information for the geographical distribution of tick species and tick-borne bacteria in Thailand and can therefore be applied for ecotourism management. © 2015 The Society for Vector Ecology.

Quantitation of 4-Methyl-4-sulfanylpentan-2-one (4MSP) in Hops by a Stable Isotope Dilution Assay in Combination with GC×GC-TOFMS: Method Development and Application To Study the Influence of Variety, Provenance, Harvest Year, and Processing on 4MSP Concentrations.

Science.gov (United States)

Reglitz, Klaas; Steinhaus, Martin

2017-03-22

A stable isotope dilution assay was developed for quantitation of 4-methyl-4-sulfanylpentan-2-one (4MSP) in hops. The approach included the use of 4-( 13 C)methyl-4-sulfanyl(1,3,5- 13 C 3 )pentan-2-one as internal standard, selective isolation of hop thiols by mercurated agarose, and GC×GC-TOFMS analysis. Application of the method to 53 different hop samples revealed 4MSP concentrations between <1 and 114 μg/kg. Notably high concentrations were associated with United States varieties such as Citra, Eureka, Simcoe, and Apollo, whereas 4MSP was absent from traditional German and English varieties. Further experiments showed that besides the variety, also harvest year and storage vitally influenced 4MSP concentrations, whereas the impact of provenance was less pronounced. Hop processing such as drying and pelletizing had only a minor impact on 4MSP concentrations. Like the majority of other hop volatiles, 4MSP is predominantly located in the lupulin glands.

Identificación de un epítope conformacional en el extremo carboxilo de la proteína MSP-1 de P. Falciparum

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Julio C. Calvo

2010-11-01

Full Text Available El fragmento carboxilo terminal, rico en cisteínas, de la proteína 1 de superficie del merozoito (MSP-1 desempeña un papel importante en la invasión del merozoito de la malaria al eritrocito. Anticuerpos contra esta región de MSP-l inhiben la invasión.

Estimated seroprevalence of Anaplasma spp. and spotted fever group Rickettsia exposure among herders and livestock in Mongolia.

Science.gov (United States)

von Fricken, Michael E; Lkhagvatseren, Sukhbaatar; Boldbaatar, Bazartseren; Nymadawa, Pagbajab; Weppelmann, Thomas A; Baigalmaa, Bekh-Ochir; Anderson, Benjamin D; Reller, Megan E; Lantos, Paul M; Gray, Gregory C

2018-01-01

To better understand the epidemiology of tick-borne disease in Mongolia, a comprehensive seroprevalence study was conducted investigating exposure to Anaplasma spp. and spotted fever group (SFG) Rickettsia spp. in nomadic herders and their livestock across three provinces from 2014 to 2015. Blood was collected from 397 herders and 2370 livestock, including sheep, goats, cattle, horses and camels. Antibodies against Anaplasma spp. and SFG Rickettsia were determined by indirect immunofluorescence using commercially available slides coated with Anaplasma phagocytophilum and Rickettsia rickettsii antigens. Logistic regression was used to determine if the odds of previous exposure differed by gender, location, and species, with or without adjustment for age. To examine the association between seroprevalence and environmental variables we used ArcGIS to circumscribe the five major clusters where human and animal data were collected. Anaplasma spp. exposure was detected in 37.3% (136/365) of humans and 47.3% (1120/2370) of livestock; SFG Rickettsia exposure was detected in 19.5% (73/374) humans and 20.4% (478/2342) livestock. Compared to the southern province (aimag) of Dornogovi, located in the Gobi Desert, humans were significantly more likely to be exposed to Anaplasma spp. and SFG Rickettsia in the northern provinces of Tov (OR=7.3, 95% CI: 3.5, 15.1; OR=3.3, 95% CI: 1.7, 7.5), and Selenge (OR=6.9, 95% CI: 3.4, 14.0; OR=2.2, 95% CI: 1.1, 4.8). The high seroprevalence of Anaplasma spp. and SFG Rickettsia in humans and livestock suggests that exposure to tick-borne pathogens may be common in herders and livestock in Mongolia, particularly in the more northern regions of the country. Until more is known about these pathogens in Mongolia, physicians and veterinarians in the countryside should consider testing for Anaplasma and SFG Rickettsia infections and treating clinically compatible cases, while public health authorities should expand surveillance efforts for these

Modular Integrated Stackable Layers (MISL) MI_MSP430A Board Design Document (BDD)

Science.gov (United States)

Yim, Hester

2013-01-01

This is a board-level design document for Modular Integrated Stackable Layers (MISL) MI_MSP430A board (PIN MSP430F5438A). The Board Design Document (BDD) contains the description, features of microcontroller, electrical and mechanical design, and drawings.

Characterization of AlgMsp, an alginate lyase from Microbulbifer sp. 6532A.

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Steven M Swift

Full Text Available Alginate is a polysaccharide produced by certain seaweeds and bacteria that consists of mannuronic acid and guluronic acid residues. Seaweed alginate is used in food and industrial chemical processes, while the biosynthesis of bacterial alginate is associated with pathogenic Pseudomonas aeruginosa. Alginate lyases cleave this polysaccharide into short oligo-uronates and thus have the potential to be utilized for both industrial and medicinal applications. An alginate lyase gene, algMsp, from Microbulbifer sp. 6532A, was synthesized as an E.coli codon-optimized clone. The resulting 37 kDa recombinant protein, AlgMsp, was expressed, purified and characterized. The alginate lyase displayed highest activity at pH 8 and 0.2 M NaCl. Activity of the alginate lyase was greatest at 50°C; however the enzyme was not stable over time when incubated at 50°C. The alginate lyase was still highly active at 25°C and displayed little or no loss of activity after 24 hours at 25°C. The activity of AlgMsp was not dependent on the presence of divalent cations. Comparing activity of the lyase against polymannuronic acid and polyguluronic acid substrates showed a higher turnover rate for polymannuronic acid. However, AlgMSP exhibited greater catalytic efficiency with the polyguluronic acid substrate. Prolonged AlgMsp-mediated degradation of alginate produced dimer, trimer, tetramer, and pentamer oligo-uronates.

Molecular approaches to detect and study the organisms causing ...

African Journals Online (AJOL)

This review will summarise the molecular approaches used to detect and analyse the genomes of Babesia bovis, B. bigemina and Anaplasma marginale which cause bovine babesiosis and anaplasmosis. These tick borne diseases are widely distributed in Africa, Asia, Australia, and Central and South America and for ...

Molecular and phylogenetic analysis of Anaplasma spp. in sheep and goats from six provinces of China.

Science.gov (United States)

Zhang, Yan; Lv, Yali; Zhang, Feifei; Zhang, Wenjing; Wang, Jinhong; Cui, Yanyan; Wang, Rongjun; Jian, Fuchun; Zhang, Longxian; Ning, Changshen

2016-12-30

Members of the genus Anaplasma are important emerging tick-borne pathogens in both humans and animals in tropical and subtropical areas. Here, we investigated the presence of Anaplasma spp. in 621 sheep and 710 goats from six provinces of China. Polymerase chain reaction (PCR) and DNA sequencing were conducted to determine the prevalence of Anaplasma (A.) phagocytophilum, A. ovis and A. bovis targeting the 16S ribosomal RNA or the major surface protein 4 gene. PCR revealed Anaplasma in 39.0% (240/621) of sheep and 45.5% (323/710) of goats. The most frequently detected species was A. ovis (88/621, 14.2% for sheep; 129/710, 18.2% for goats), followed by A. bovis (60/621, 9.7% for sheep; 74/710, 10.4% for goats) and A. phagocytophilum (33/621, 5.3% for sheep; 15/710, 2.1% for goats). Additionally, eight sheep and 20 goats were found to be infected with three pathogens simultaneously. DNA sequencing confirmed the presence of these three Anaplasma species in the investigated areas, and phylogenetic analysis indicated that there was geographic segregation to a certain extent, as well as a relationship between the host and cluster of A. ovis. The results of the present study provide valuable data that helps understand the epidemiology of anaplasmosis in ruminants from China.

Cellular responses to modified Plasmodium falciparum MSP119 antigens in individuals previously exposed to natural malaria infection

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Awobode Henrietta O

2009-11-01

Full Text Available Abstract Background MSP1 processing-inhibitory antibodies bind to epitopes on the 19 kDa C-terminal region of the Plasmodium falciparum merozoite surface protein 1 (MSP119, inhibiting erythrocyte invasion. Blocking antibodies also bind to this antigen but prevent inhibitory antibodies binding, allowing invasion to proceed. Recombinant MSP119 had been modified previously to allow inhibitory but not blocking antibodies to continue to bind. Immunization with these modified proteins, therefore, has the potential to induce more effective protective antibodies. However, it was unclear whether the modification of MSP119 would affect critical T-cell responses to epitopes in this antigen. Methods The cellular responses to wild-type MSP119 and a panel of modified MSP119 antigens were measured using an in-vitro assay for two groups of individuals: the first were malaria-naïve and the second had been naturally exposed to Plasmodium falciparum infection. The cellular responses to the modified proteins were examined using cells from malaria-exposed infants and adults. Results Interestingly, stimulation indices (SI for responses induced by some of the modified proteins were at least two-fold higher than those elicited by the wild-type MSP119. A protein with four amino acid substitutions (Glu27→Tyr, Leu31→Arg, Tyr34→Ser and Glu43→Leu had the highest stimulation index (SI up to 360 and induced large responses in 64% of the samples that had significant cellular responses to the modified proteins. Conclusion This study suggests that specific MSP119 variants that have been engineered to improve their antigenicity for inhibitory antibodies, retain T-cell epitopes and the ability to induce cellular responses. These proteins are candidates for the development of MSP1-based malaria vaccines.

Bovine babesiosis and anaplasmosis complex: diagnosis and evaluation of the risk factors from Bahia, Brazil

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Lucimar Souza Amorim

Full Text Available Direct diagnoses were made by using - blood smears and nested PCR (nPCR tests on 309 blood samples from crossbred dairy cattle in the municipality of Ibicaraí, Bahia. From diagnostic blood smear slides, the observed parasitic frequencies were 31.1% for Anaplasma marginale and 20.4% for Babesia sp. From nPCR diagnoses, they were 63% for A. marginale, 34% for Babesia bigemina and 20.4% for Babesia bovis. There were significant differences (P <0.01 between the two diagnostic methods (nPCR and blood smear slides. The compliance obtained from the kappa test was 0.41 and 0.48 for A. marginale and Babesia sp., respectively. The tick samples from the six farms analyzed using nPCR were only positive for A. marginale. Evaluation of the risk factors relating to the presence of ticks and the age of the animals showed that there was a significant association (P <0.01 with the frequency of animals infected with both pathogens. Therefore, under the conditions studied, nPCR proved to be a good tool for diagnosing the agents of the bovine babesiosis and anaplasmosis complex because of its sensitivity and specificity in comparison with blood smears. The municipality of Ibicaraí is an area with endemic prevalence of bovine babesiosis and anaplasmosis confirmed by nPCR and A. marginale is the main agent of the disease.

Seroprevalence and risk factors associated with Ehrlichia canis, Anaplasma spp., Borrelia burgdorferi sensu lato, and D. immitis in hunting dogs from southern Italy.

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Piantedosi, Diego; Neola, Benedetto; D'Alessio, Nicola; Di Prisco, Francesca; Santoro, Mario; Pacifico, Laura; Sgroi, Giovanni; Auletta, Luigi; Buch, Jesse; Chandrashekar, Ramaswamy; Breitschwerdt, Edward B; Veneziano, Vincenzo

2017-10-01

Canine vector-borne diseases (CVBDs) are caused by a range of pathogens transmitted to dogs by arthropods. The present study investigates Ehrlichia canis, Anaplasma spp., Borrelia burgdorferi sensu lato, and Dirofilaria immitis seroprevalences in hunting dogs from southern Italy. Dogs (no. 1335) were tested using a commercial in-clinic enzyme-linked immunosorbent assay kit. Odds ratios (ORs) were calculated by logistic regression analysis to identify risk factors. Overall, 138/1335 dogs (10.3%) were seroreactive to at least one CVBD pathogen. E. canis, Anaplasma spp., B. burgdorferi s.l., and D. immitis seroprevalences were 7.6, 4.4, 0.3, and 0.2%, respectively. E. canis and Anaplasma spp. co-exposures were found in 30 dogs (2.2%), compared with Anaplasma spp. and B. burgdorferi s.l. co-exposures in 2 dogs (0.1%). Adult age was a risk factor for E. canis (OR 2.35) seroreactivity whereas hunting fur-bearing animals for E. canis (OR 4.75) and Anaplasma spp. (OR 1.87), respectively. The historical presence of tick infestation was identified as a risk factor for positivity to E. canis (OR 2.08) and Anaplasma spp. (OR 2.15). Finally, larger dog pack size was significantly associated with E. canis (OR 1.85) and Anaplasma spp. (OR 2.42) exposures. The results of the present survey indicated that hunting dog populations are at relative risk of CVBDs in southern Italy. Further studies are needed to evaluate the role of hunting dogs in the epidemiology of vector-borne organisms due to sharing common environments with wild, sympatric animal populations.

Impact of pre-existing MSP142-allele specific immunity on potency of an erythrocytic Plasmodium falciparum vaccine

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Bergmann-Leitner Elke S

2012-09-01

Full Text Available Abstract Background MSP1 is the major surface protein on merozoites and a prime candidate for a blood stage malaria vaccine. Preclinical and seroepidemiological studies have implicated antibodies to MSP1 in protection against blood stage parasitaemia and/or reduced parasite densities, respectively. Malaria endemic areas have multiple strains of Plasmodium falciparum circulating at any given time, giving rise to complex immune responses, an issue which is generally not addressed in clinical trials conducted in non-endemic areas. A lack of understanding of the effect of pre-existing immunity to heterologous parasite strains may significantly contribute to vaccine failure in the field. The purpose of this study was to model the effect of pre-existing immunity to MSP142 on the immunogenicity of blood-stage malaria vaccines based on alternative MSP1 alleles. Methods Inbred and outbred mice were immunized with various recombinant P. falciparum MSP142 proteins that represent the two major alleles of MSP142, MAD20 (3D7 and Wellcome (K1, FVO. Humoral immune responses were analysed by ELISA and LuminexTM, and functional activity of induced MSP142-specific antibodies was assessed by growth inhibition assays. T-cell responses were characterized using ex vivo ELISpot assays. Results Analysis of the immune responses induced by various immunization regimens demonstrated a strong allele-specific response at the T cell level in both inbred and outbred mice. The success of heterologous regimens depended on the degree of homology of the N-terminal p33 portion of the MSP142, likely due to the fact that most T cell epitopes reside in this part of the molecule. Analysis of humoral immune responses revealed a marked cross-reactivity between the alleles. Functional analyses showed that some of the heterologous regimens induced antibodies with improved growth inhibitory activities. Conclusion The development of a more broadly efficacious MSP1 based vaccine may be

The Life and Death of Anaplasma

Centers for Disease Control (CDC) Podcasts

Dr. Setu Vora, medical director of critical care and physician director of performance improvement at Backus Hospital in Norwich, Connecticut, reads his poem The Life and Death of Anaplasma and discusses the poem’s origins.

Diagnosis of hemoparasitosis trough the spread-smear technique using a drop of blood clot

OpenAIRE

Neusa Saltiél Stobbe; Eunice Leonora Chaplin; Maria das Graças de Souza Paiva; Nilton Rogério Santos Silva; Flávio A. Pacheco Araújo; Elinor Fortes

1992-01-01

The spread-smear technique using a drop of blood clot and stained by diluted and undimitcd Giemsa stain was evaluated for the search of blood cell types and recognition of Babesia bovis, Babesia bigemina and Anaplasma marginale hemoparasites. The blood clot samples were taken from a 6 month-old calf experimentally inoculated with the blood from a Cattle Tick Fever carrier. The Technique was sumitable for the observation of different blood cells and for the recognition of the hemoparasites.

Broadly reactive antibodies specific for Plasmodium falciparum MSP-119 are associated with the protection of naturally exposed children against infection

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Dent Arlene E

2012-08-01

Full Text Available Abstract Background The 19 kDa C-terminal region of Plasmodium falciparum Merozoite Surface Protein-1 is a known target of naturally acquired humoral immunity and a malaria vaccine candidate. MSP-119 has four predominant haplotypes resulting in amino acid changes labelled EKNG, QKNG, QTSR and ETSR. IgG antibodies directed against all four variants have been detected, but it is not known if these variant specific antibodies are associated with haplotype-specific protection from infection. Methods Blood samples from 201 healthy Kenyan adults and children who participated in a 12-week treatment time-to-infection study were evaluated. Venous blood drawn at baseline (week 0 was examined for functional and serologic antibodies to MSP-119 and MSP-142 variants. MSP-119 haplotypes were detected by a multiplex PCR assay at baseline and weekly throughout the study. Generalized linear models controlling for age, baseline MSP-119 haplotype and parasite density were used to determine the relationship between infecting P. falciparum MSP-119 haplotype and variant-specific antibodies. Results A total of 964 infections resulting in 1,533 MSP-119 haplotypes detected were examined. The most common haplotypes were EKNG and QKNG, followed by ETSR and QTSR. Children had higher parasite densities, greater complexity of infection (>1 haplotype, and more frequent changes in haplotypes over time compared to adults. Infecting MSP-119 haplotype at baseline (week 0 had no influence on haplotypes detected over the subsequent 11 weeks among children or adults. Children but not adults with MSP-119 and some MSP-142 variant antibodies detected by serology at baseline had delayed time-to-infection. There was no significant association of variant-specific serology or functional antibodies at baseline with infecting haplotype at baseline or during 11 weeks of follow up among children or adults. Conclusions Variant transcending IgG antibodies to MSP-119 are associated with protection

Molecular detection of Rickettsia, Anaplasma, Coxiella and Francisella bacteria in ticks collected from Artiodactyla in Thailand.

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Sumrandee, Chalao; Baimai, Visut; Trinachartvanit, Wachareeporn; Ahantarig, Arunee

2016-07-01

A total of 79 ticks collected from Sambar deer (Cervus unicolor), Barking deer (Muntiacus muntjak) and Wild boar (Sus scrofa) were examined by PCR for the presence of Rickettsia, Anaplasma, Coxiella, and Francisella bacteria. Of the 79 ticks, 13% tested positive for Rickettsia, 15% tested positive for Anaplasma, 4% tested positive for Coxiella, and 3% tested positive for Francisella. Interestingly, triple infection with Anaplasma, Rickettsia and Francisella was determined in a Dermacentor auratus tick. Moreover, another triple infection with Rickettsia, Anaplasma, and Coxiella was found in a Haemaphysalis lagrangei tick. Double infection of Rickettsia with Coxiella was also detected in another H. lagrangei tick. From the phylogenetic analyses, we found a Rickettsia sp. with a close evolutionary relationship to Rickettsia bellii in the H. lagrangei tick. We also found the first evidence of a Rickettsia sp. that is closely related to Rickettsia tamurae in Rhipicephalus (Boophilus) microplus ticks from Thailand. H. lagrangei and Haemaphysalis obesa ticks collected from Sambar deer tested positive for Anaplasma species form the same clade with Anaplasma bovis. In contrast, other H. lagrangei ticks collected from Sambar deer and D. auratus ticks collected from Wild boar were also reported for the first time to be infected with an Anaplasma species that is closely related to Anaplasma platys. The phylogenetic analysis of the 16S rRNA gene of Coxiella bacteria revealed that Coxiella symbionts from H. lagrangei formed a distinctly different lineage from Coxiella burnetii (a human pathogen). Additionally, Francisella bacteria identified in D. auratus ticks were found to be distantly related to a group of pathogenic Francisella species. The identification of these bacteria in several feeding ticks suggests the risk of various emerging tick-borne diseases and endosymbionts in humans, wildlife, and domestic animals in Thailand. Copyright © 2016 Elsevier GmbH. All rights

Construcción de Distribuciones Multivariadas con Marginales Dependientes Usando Cópulas en R

OpenAIRE

Jaramillo-Elorza, M. C; Lozano, J. A

2014-01-01

Las cópulas se han convertido en una herramienta popular para la construcción de modelos multivariados en campos donde la dependencia multivariada es de gran interés. El propósito de este trabajo es presentar las cópulas tanto en su concepto teórico, como en su implementación en el software estadístico R y profundizar en la construcción de distribuciones multivariadas con marginales dependientes, usando la clase mvdc del paquete copula, la cual permite utilizar varias y diferentes marginales ...

[Natural infection by hemoparasites in calves submitted to chemoprophylaxis at 30 days of age].

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da Silva, Rosângela A; Corrêa, Fabíola do N; Botteon, Rita de Cássia C M; Botteon, Paulo de Tarso L

2007-01-01

The tick-borne disease (TBD) brings great damages to cattle breeding. The most important etiologic agents are Babesia bigemina, B. bovis and Anaplasma marginale, being the tick Boophilus microplus the main vector. This work reports the occurrence of natural infection by hemoparasites of TBD in 36 calves with high ticks natural infestation submitted to chemoprophylaxis with 30 days year-old. The blood smears from animals of different ages were analized and were found B. bigemina (33.3%), B. bovis (11.1%) and A. marginale (13.9%). Six animals had clinical symptoms (16.7%) and one dead (2.8%). The number of clinical cases ocurred in consequence of an association of factors as high infestation of ticks and low passive immunity in period that calves had not developed enough active immunity.

Isolation and Characterization of Vaccine Candidate Genes Including CSP and MSP1 in Plasmodium yoelii.

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Kim, Seon-Hee; Bae, Young-An; Seoh, Ju-Young; Yang, Hyun-Jong

2017-06-01

Malaria is an infectious disease affecting humans, which is transmitted by the bite of Anopheles mosquitoes harboring sporozoites of parasitic protozoans belonging to the genus Plasmodium . Despite past achievements to control the protozoan disease, malaria still remains a significant health threat up to now. In this study, we cloned and characterized the full-unit Plasmodium yoelii genes encoding merozoite surface protein 1 (MSP1), circumsporozoite protein (CSP), and Duffy-binding protein (DBP), each of which can be applied for investigations to obtain potent protective vaccines in the rodent malaria model, due to their specific expression patterns during the parasite life cycle. Recombinant fragments corresponding to the middle and C-terminal regions of PyMSP1 and PyCSP, respectively, displayed strong reactivity against P. yoelii -infected mice sera. Specific native antigens invoking strong humoral immune response during the primary and secondary infections of P. yoelii were also abundantly detected in experimental ICR mice. The low or negligible parasitemia observed in the secondary infected mice was likely to result from the neutralizing action of the protective antibodies. Identification of these antigenic proteins might provide the necessary information and means to characterize additional vaccine candidate antigens, selected solely on their ability to produce the protective antibodies.

HEMOPARASITOSIS EN GANADERÍA DOBLE PROPÓSITO VENEZOLANA, DIAGNÓSTICO Y CONTROL: UNA REVISIÓN

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Rita Tamasaukas

2010-01-01

Full Text Available Este trabajo de revisión de tesis doctoral incluye resultados de investigaciones realizadas en el periodo 1971 a 2009 sobre cuatro hemoparasitosis causados por protozoarios y rickettsiales en Venezuela y otros países, con énfasis en la infección mixta en rebaños bovinos, y su diagnóstico y control. En Venezuela, las afecciones causadas por agentes hemotrópicos parasitarios están distribuidas en todos los estados del país con vocación ganadera, en especial, en rebaños bovinos y bufalinos. Entre aquellos encontramos a especies del género Anaplasma (Anaplasma marginale, Babesia (Babesia bigemina y B. bovis, y Trypanosoma (Trypanosoma vivax, con una frecuencia endémica y con variaciones estacionales.

Molecular epidemiology of the emerging zoonosis agent Anaplasma phagocytophilum (Foggie, 1949) in dogs and ixodid ticks in Brazil.

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Santos, Huarrisson A; Thomé, Sandra M G; Baldani, Cristiane D; Silva, Claudia B; Peixoto, Maristela P; Pires, Marcus S; Vitari, Gabriela L V; Costa, Renata L; Santos, Tiago M; Angelo, Isabele C; Santos, Leandro A; Faccini, João L H; Massard, Carlos L

2013-12-11

Anaplasma phagocytophilum is an emerging pathogen of humans, dogs and other animals, and it is transmitted by ixodid ticks. The objective of the current study was a) detect A. phagocytophilum in dogs and ixodid ticks using real-time Polymerase Chain Reaction (qPCR); and b) Determine important variables associated to host, environment and potential tick vectors that are related to the presence of A. phagocytophilum in dogs domiciled in Rio de Janeiro, Brazil. We tested blood samples from 398 dogs and samples from 235 ticks, including 194 Rhipicephalus sanguineus sensu lato, 15 Amblyomma cajennense, 8 Amblyomma ovale and 18 pools of Amblyomma sp. nymphs. A semi-structured questionnaire was applied by interviewing each dog owner. Deoxyribonucleic acid obtained from ticks and dog buffy coat samples were amplified by qPCR (msp2 gene). The sequencing of 16S rRNA and groESL heat shock operon genes and a phylogenetic analysis was performed. The multiple logistic regression model was created as a function of testing positive dogs for A. phagocytophilum. Among the 398 blood samples from dogs, 6.03% were positive for A. phagocytophilum. Anaplasma phagocytophilum was detected in one A. cajennense female tick and in five R. sanguineus sensu lato ticks (four males and one female). The partial sequences of the 16S rRNA, and groESL genes obtained were highly similar to strains of A. phagocytophilum isolated from wild birds from Brazil and human pathogenic strains. The tick species collected in positive dogs were R. sanguineus sensu lato and A. cajennense, with A.cajennense being predominant. Tick infestation history (OR = 2.86, CI = 1.98-14.87), dog size (OR = 2.41, IC: 1.51-12.67), the access to forest areas (OR = 3:51, CI: 1.52-16.32), hygiene conditions of the environment in which the dogs lived (OR = 4.35, CI: 1.86-18.63) and Amblyomma sp. infestation (OR = 6.12; CI: 2.11-28.15) were associated with A. phagocytophilum infection in dogs. This is the

Retrospective study of hemoparasites in cattle in southern Italy by reverse line blot hybridization.

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Ceci, Luigi; Iarussi, Fabrizio; Greco, Beatrice; Lacinio, Rosanna; Fornelli, Stefania; Carelli, Grazia

2014-06-01

Tick-borne diseases are widespread in tropical and temperate regions and are responsible for important economic losses in those areas. In order to assess the presence and prevalence of various pathogens in southern Italy, we retrospectively analyzed cattle blood samples collected for a previous study in 2000 using reverse line blot (RLB) hybridization. The study had been carried out in three regions of southern Italy on 1,500 randomly selected and apparently healthy adult cattle. RLB showed that 43.7% of the cattle were positive for nine different species of hemoparasites with either a single infection or a mixed infection. Theileria buffeli was the most common species found, being present in 27.3% of the animals, followed by Anaplasma marginale in 18.1%, Anaplasma centrale in 13.8%, Babesia bigemina and Anaplasma bovis in 4.2%, Anaplasma phagocytophilum in 1.7%, Babesia bovis in 1.6%, Babesia major in 0.2% and Babesia divergens in 0.1%. Complete blood counts showed different degrees of anemia in 363 animals (24.2%) and of these, 169 were RLB-positive for at least one pathogen. Among the ticks that were collected from the cattle, the following species were identified: Rhipicephalus bursa, Ixodes ricinus, Hyalomma marginatum, Boophilus annulatus, Dermacentor marginatus and Haemaphysalis (sulcata, parva, inermis and punctata). The results obtained confirmed the spread of endemic tick-borne pathogens in the regions studied.

Alpha proteobacteria of genus Anaplasma (Rickettsiales: Anaplasmataceae): Epidemiology and characteristics of Anaplasma species related to veterinary and public health importance.

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Atif, Farhan Ahmad

2016-05-01

The Anaplasma species are important globally distributed tick-transmitted bacteria of veterinary and public health importance. These pathogens, cause anaplasmosis in domestic and wild animal species including humans. Rhipicephalus, Ixodes, Dermacentor and Amblyomma genera of ticks are the important vectors of Anaplasma. Acute anaplasmosis is usually diagnosed upon blood smear examination followed by antibodies and nucleic acid detection. All age groups are susceptible but prevalence increases with age. Serological cross-reactivity is one of the important issues among Anaplasma species. They co-exist and concurrent infections occur in animals and ticks in same geographic area. These are closely related bacteria and share various common attributes which should be considered while developing vaccines and diagnostic assays. Movement of susceptible animals from non-endemic to endemic regions is the major risk factor of bovine/ovine anaplasmosis and tick-borne fever. Tetracyclines are currently available drugs for clearance of infection and treatment in humans and animals. Worldwide vaccine is not yet available. Identification, elimination of reservoirs, vector control (chemical and biological), endemic stability, habitat modification, rearing of tick resistant breeds, chemotherapy and tick vaccination are major control measures of animal anaplasmosis. Identification of reservoirs and minimizing the high-risk tick exposure activities are important control strategies for human granulocytic anaplasmosis.

Neonatal Anaplasma platys infection in puppies: Further evidence for possible vertical transmission

Czech Academy of Sciences Publication Activity Database

Matei, I.A.; Stuen, S.; Modrý, David; Degan, A.; D'Amico, G.; Mihalca, A. D.

2017-01-01

Roč. 219, 1 January (2017), s. 40-41 ISSN 1090-0233 Institutional support: RVO:60077344 Keywords : Anaplasma platys * canine * vertical transmission Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine OBOR OECD: Veterinary science Impact factor: 1.802, year: 2016

Diagnosis of hemoparasitosis trough the spread-smear technique using a drop of blood clot

Directory of Open Access Journals (Sweden)

Neusa Saltiél Stobbe

1992-12-01

Full Text Available The spread-smear technique using a drop of blood clot and stained by diluted and undimitcd Giemsa stain was evaluated for the search of blood cell types and recognition of Babesia bovis, Babesia bigemina and Anaplasma marginale hemoparasites. The blood clot samples were taken from a 6 month-old calf experimentally inoculated with the blood from a Cattle Tick Fever carrier. The Technique was sumitable for the observation of different blood cells and for the recognition of the hemoparasites.

Naturally-acquired humoral immune responses against the N- and C-termini of the Plasmodium vivax MSP1 protein in endemic regions of Brazil and Papua New Guinea using a multiplex assay

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Alonso Pedro L

2010-01-01

Full Text Available Abstract Background Progress towards the development of a malaria vaccine against Plasmodium vivax, the most widely distributed human malaria parasite, will require a better understanding of the immune responses that confer clinical protection to patients in regions where malaria is endemic. Methods Glutathione S-transferase (GST and GST-fusion proteins representing the N- terminus of the merozoite surface protein 1 of P. vivax, PvMSP1-N, and the C-terminus, PvMSP1-C, were covalently coupled to BioPlex carboxylated beads. Recombinant proteins and coupled beads were used, respectively, in ELISA and Bioplex assays using immune sera of P. vivax patients from Brazil and PNG to determine IgG and subclass responses. Concordances between the two methods in the seropositivity responses were evaluated using the Kappa statistic and the Spearman's rank correlation. Results The results using this methodology were compared with the classical microtitre enzyme-linked immnosorbent assay (ELISA, showing that the assay was sensitive, reproducible and had good concordance with ELISA; yet, further research into different statistical analyses seems desirable before claiming conclusive results exclusively based on multiplex assays. As expected, results demonstrated that PvMSP1 was immunogenic in natural infections of patients from different endemic regions of Brazil and Papua New Guinea (PNG, and that age correlated only with antibodies against the C-terminus part of the molecule. Furthermore, the IgG subclass profiles were different in these endemic regions having IgG3 predominantly recognizing PvMSP1 in Brazil and IgG1 predominantly recognizing PvMSP1 in PNG. Conclusions This study validates the use of the multiplex assay to measure naturally-acquired IgG antibodies against the merozoite surface protein 1 of P. vivax.

In situ observations of meteor smoke particles (MSP during the Geminids 2010: constraints on MSP size, work function and composition

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M. Rapp

2012-12-01

Full Text Available We present in situ observations of meteoric smoke particles (MSP obtained during three sounding rocket flights in December 2010 in the frame of the final campaign of the Norwegian-German ECOMA project (ECOMA = Existence and Charge state Of meteoric smoke particles in the Middle Atmosphere. The flights were conducted before, at the maximum activity, and after the decline of the Geminids which is one of the major meteor showers over the year. Measurements with the ECOMA particle detector yield both profiles of naturally charged particles (Faraday cup measurement as well as profiles of photoelectrons emitted by the MSPs due to their irradiation by photons of a xenon-flash lamp. The column density of negatively charged MSPs decreased steadily from flight to flight which is in agreement with a corresponding decrease of the sporadic meteor flux recorded during the same period. This implies that the sporadic meteors are a major source of MSPs while the additional influx due to the shower meteors apparently did not play any significant role. Surprisingly, the profiles of photoelectrons are only partly compatible with this observation: while the photoelectron current profiles obtained during the first and third flight of the campaign showed a qualitatively similar behaviour as the MSP charge density data, the profile from the second flight (i.e., at the peak of the Geminids shows much smaller photoelectron currents. This may tentatively be interpreted as a different MSP composition (and, hence, different photoelectric properties during this second flight, but at this stage we are not in a position to conclude that there is a cause and effect relation between the Geminids and this observation. Finally, the ECOMA particle detector used during the first and third flight employed three instead of only one xenon flash lamp where each of the three lamps used for one flight had a different window material resulting in different cut off wavelengths for these

Remodeling of tick cytoskeleton in response to infection with Anaplasma phagocytophilum

Czech Academy of Sciences Publication Activity Database

Cabezas Cruz, Alejandro; Alberdi, P.; Valdés, James J.; Villar, M.; de la Fuente, J.

2017-01-01

Roč. 22, Jun 1 (2017), s. 1830-1844 ISSN 1093-4715 EU Projects: European Commission(XE) 278976 - ANTIGONE Institutional support: RVO:60077344 Keywords : cytoskeleton * proteomics * transcriptomics * Ixodes scapularis * Anaplasma phagocytophilum Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Biochemistry and molecular biology

Molecular Characterization and Phylogenetic Analysis of Anaplasma spp. and Ehrlichia spp. Isolated from Various Ticks in Southeastern and Northwestern Regions of Iran.

Science.gov (United States)

Jafar Bekloo, Ahmad; Ramzgouyan, Maryam Roya; Shirian, Sadegh; Faghihi, Faezeh; Bakhshi, Hassan; Naseri, Fatemeh; Sedaghat, Mehdi; Telmadarraiy, Zakkyeh

2018-05-01

Anaplasma/Ehrlichia species are tick-transmitted pathogens that cause infections in humans and numerous domestic and wild animal species. There is no information available on the molecular characteristics and phylogenetic position of Anaplasma/Ehrlichia spp. isolated from tick species from different geographic locations in Iran. The aim of this study was to determine the prevalence, molecular characteristics, and phylogenetic relationship of both Anaplasma spp. and Ehrlichia spp. in tick species isolated from different domestic animals from two different geographical locations of Iran. A total of 930 ticks were collected from 93 cattle, 250 sheep, and 587 goats inhabiting the study areas. The collected ticks were then investigated for the presence of Anaplasma/Ehrlichia spp. using nested PCR based on the 16S rRNA gene, followed by sequencing. Sequence analysis was done based on the data published in the GenBank on Anaplasma/Ehrlichia spp. isolates using bioinformatic tools such as the standard nucleotide BLAST. Genome of Anaplasma or Ehrlichia spp. was detected in 14 ticks collected in Heris, including 5 Dermacentor marginatus, 1 Haemaphysalis erinacei, 3 Hyalomma anatolicum, and 4 Rhipicephalus sanguineus, also in 29 ticks collected in Chabahar, including 14 R. sanguineus, 8 D. marginatus, 3 Hyalomma Anatolicum, and 4 Hyalomma dromedarii. Partial analysis of the 16S rRNA gene sequence of positive samples collected from goats and sheep showed that they were infected with Anaplasma/Ehrlichia spp. that were 94-98% identical to ovine Anaplasma and 91-96% identical to Neoehrlichia and Ehrlichia spp. The various ticks identified in this study suggest the possible emergence of tick-borne diseases in animals and humans in these regions. R. sanguineus and D. marginatus seem to be predominant vectors responsible for anaplasmosis in these regions. Partial sequence analysis of the 16S rRNA gene showed that A. ovis is genetically polymorphic in these regions. Furthermore, an

Insulin gene VNTR polymorphisms -2221MspI and -23HphI are associated with type 1 diabetes and latent autoimmune diabetes in adults: a meta-analysis.

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Zhang, Na; Huang, Weihuang; Dong, Fang; Liu, Yang; Zhang, Baohuan; Jing, Lipeng; Wang, Man; Yang, Guang; Jing, Chunxia

2015-12-01

A variable number of tandem repeat (VNTRs) region in the insulin gene (INS) possibly influences the progression of type 1 diabetes (T1D) and latent autoimmune diabetes in adults (LADA). However, effects of INS VNTR polymorphisms in these contexts remain inconclusive. We performed a systematic review of work on the INS VNTR -2221MspI and -23HphI polymorphisms to estimate the overall effects thereof on disease susceptibility; we included 17,498 T1D patients and 24,437 controls, and 1960 LADA patients and 5583 controls. For T1D, the C allele at -2221MspI and the A allele at -23HphI were associated with estimated relative risks of 2.13 (95 % CI 1.94, 2.35) and 0.46 (95 % CI 0.44, 0.48), which contributed to absolute increases of 46.76 and 46.98 % in the risk of all T1D, respectively. The estimated lambda values were 0.44 and 0.42, respectively, suggesting that a co-dominant model most likely explained the effects of -2221MspI and -23HphI on T1D. For -23HphI, the A allele carried an estimated relative risk of 0.55 (95 % CI 0.50, 0.61) for LADA and increased the risk of all LADA by 36.94 %. The λ value was 0.43, suggesting that a co-dominant model most likely explained the effect of -23HphI on LADA. Our results support the existence of associations of INS with T1D and LADA.

FAST's Discovery of a New Millisecond Pulsar (MSP) toward the Fermi-LAT unassociated source 3FGL J0318.1+0252

Science.gov (United States)

Wang, Pei; Li, Di; Zhu, Weiwei; Zhang, Chengmin; Yan, Jun; Hou, Xian; Clark, Colin J.; Saz Parkinson, Pablo M.; Michelson, Peter F.; Ferrara, Elizabeth C.; Thompson, David J.; Smith, David A.; Ray, Paul S.; Kerr, Matthew; Shen, Zhiqiang; Wang, Na; Fermi-LAT Collaboration

2018-04-01

The Five hundred-meter Aperture Spherical radio Telescope (FAST), operated by the National Astronomical Observatories, Chinese Academy of Sciences, has discovered a radio millisecond pulsar (MSP) coincident with the unassociated gamma-ray source 3FGL J0318.1+0252 (Acero et al. 2015 ApJS, 218, 23), also known as FL8Y J0318.2+0254 in the recently released Fermi Large Area Telescope (LAT) 8-year Point Source List (FL8Y).

Producción y caracterización de un anticuerpo monoclonal que reconoce el fragmento 28-30 kDa de la proteína MSP-1 de Plasmodium falciparum

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Nina M. Muñoz

2000-06-01

Full Text Available Los merozoitos del parásito Plasmodium falciparum contienen en su superficie los fragmentos producidos por el primer procesamiento proteolítico de la proteina MSP-1 (merozoite surface protein-1. Dichos fragmentos tienen pesos moleculares relativos aproximados de 83, 42, 38 y 28-30 kDa, respectivamente. En este estudio se describe la producción y caracterización de un anticuerpo monoclonal, denominado 8F4, que reconoce la proteina MSP-1 dadas las siguientes características del antigeno: 1 localización subcelular, 2 peso molecular relativo, 3 período de expresión durante el ciclo eritrocítrico, 4 tipo de asociación con la membrana plasmática del parásito y 5 presencia de epítopes compartidos con MSP-1. Adicionalmente. se estableció que 8F4 reconoce el fragmento de 28-30 kDa, producto del primer procesamiento proteolitico de MSP-1. Los análisis realizados permiten confirmar que MSP-1, 28-30 , no permanece en la superficie del merozoito desoués de ocurrida la invasión. lo cual suoiere oue hace oarte del com.o le.io de polipéptidos que es liberado por el merozoito previamente al evento de la invasión. 8F4 es el primer anticuerpo monoclonal reportado hasta el momento que reconoce específicamente este fragmento.

Role of a genetic polymorphism in the corticotropin-releasing factor receptor 1 gene in alcohol drinking and seeking behaviors of Marchigian Sardinian alcohol-preferring (msP rats

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Lydia Ojonemile Ayanwuyi

2013-04-01

Full Text Available Marchigian Sardinian alcohol-preferring (msP rats exhibit innate preference for alcohol, are highly sensitive to stress and stress-induced alcohol seeking. Genetic analysis showed that over-expression of the corticotropin-releasing factor (CRF system of msP rats is correlated with the presence of two single nucleotide polymorphisms (SNPs occurring in the promoter region (position -1836 and -2097 of the CRF1 receptor (CRF1-R gene. Here we examined whether these point mutations were associated to the innate alcohol preference, stress-induced drinking and seeking.We have recently re-derived the msP rats to obtain two distinct lines carrying the wild type (GG and the point mutations (AA, respectively. The phenotypic characteristics of these two lines were compared with those of unselected Wistar rats. Both AA and GG rats showed similar patterns of voluntary alcohol intake and preference. Similarly, the pharmacological stressor yohimbine (0.0, 0.625, 1.25 and 2.5 mg/kg elicited increased operant alcohol self-administration under fixed and progressive ratio reinforcement schedules in all three lines. Following extinction, yohimbine (0.0, 0.625, 1.25 and 2.5 mg/kg significantly reinstated alcohol seeking in the three groups. However, at the highest dose this effect was no longer evident in AA rats. Treatment with the CRF1-R antagonist antalarmin (0, 5, 10 and 20 mg/kg significantly reduced alcohol-reinforced lever pressing in the AA line (10 and 20 mg/kg while a weaker or no effect was observed in the Wistar and the GG group, respectively. Finally, antalarmin significantly reduced yohimbine-induced increase in alcohol drinking in all three groups.In conclusion, these specific SNPs in the CRF1-R gene do not seem to play a primary role in the expression of the msP excessive-drinking phenotype or stress-induced drinking but may be associated with a decreased threshold for stress-induced alcohol seeking and an increased sensitivity to the effects of

The evaluation of CLIA, RIA and MSP-ELISA for measurement of thyroid hormones

International Nuclear Information System (INIS)

Jing Hua; Li Dan; Chen Yiguang; Zhou Huiqin; Xu Liyan

2005-01-01

To compare the characteristics of chemiluminescent immunoassay (CLIA), RIA and magnetic solid phase enzyme-linked immnosorbent assay (MSP-ELISA) in measuring thyroid hormones, TT 3 , TT 4 , FT 3 , FT 4 and TSH were tested in 40 samples of human serum and in standard samples of thyroid hormones by CLIA, RIA and MSP-ELISA respectively. The linearity, relativiy, precision, recovery of these three met hods were compared. The result showed that there were no statistical differences between CLIA, RIA and MSP-ELISA in linearity and relativity. CLIA was better than RIA and MSP-ELISA in precision and accuracy. (authors)

Molecular detection of Anaplasma platys, Anaplasma phagocytophilum and Wolbachia sp. but not Ehrlichia canis in Croatian dogs.

Science.gov (United States)

Huber, Doroteja; Reil, Irena; Duvnjak, Sanja; Jurković, Daria; Lukačević, Damir; Pilat, Miroslav; Beck, Ana; Mihaljević, Željko; Vojta, Lea; Polkinghorne, Adam; Beck, Relja

2017-11-01

The bacteria Anaplasma platys, Anaplasma phagocytophilum and Ehrlichia canis are tick-borne agents that cause canine vector-borne disease. The prevalence of these pathogens in South Eastern Europe is unknown with the exception of an isolated case of A. platys detected in a dog imported into Germany from Croatia. To gain a better insight into their presence and prevalence, PCR-based screening for these bacterial pathogens was performed on domesticated dogs from different regions of Croatia. Blood samples from 1080 apparently healthy dogs from coastal and continental parts of Croatia as well as tissue samples collected from 63 deceased dogs with a history of anaemia and thrombocytopenia were collected for molecular screening by an Anaplasmataceae-specific 16S rRNA conventional PCR. Positive samples were confirmed using a second Anaplasmataceae-specific PCR assay with the PCR product sequenced for the purpose of bacterial species identification. All sequenced isolates were georeferenced and a kernel intensity estimator was used to identify clusters of greater case intensity. 42/1080 (3.8%; CI 2.7-5.0) of the healthy dogs were PCR positive for bacteria in the Anaplasmataceae. Sequencing of the 16S rRNA gene amplified from these positive samples revealed the presence of A. platys in 2.5% (CI 1.6-3.4%, 27 dogs), A. phagocytophilum in 0.3% (CI 0-0.6%, 3 dogs) and a Wolbachia endosymbiont in 1.1% (CI 0.4-1.6%, 12 dogs) of dogs screened in this study. Necropsied dogs were free from infection. Notably, no evidence of E. canis infection was found in any animal. This survey represents a rare molecular study of Anaplasmataceae in dogs in South Eastern Europe, confirming the presence of A. platys and A. phagocytophilum but not E. canis. The absence of E. canis was surprising given it has been described in all other Mediterranean countries surveyed and raises questions over the regional vector capacity of the Rhipicephalus sanguineus tick.

High infection of Anaplasma and Ehrlichia spp. among tick species collected from different geographical locations of Iran

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Leila Tajedin

2016-10-01

Full Text Available Objective: To ascertain the prevalence of the Anaplasma/Ehrlichia infections in tick population within four provinces of Iran. Methods: A total of 384 tick specimens were collected from domestic animals inhabiting in four provinces (East Azerbaijan, Gilan, South Khorasan and Yazd. Specimens were identified based on morphological analysis. The detection of Anaplasma spp./Ehrlichia spp. within tick samples was carried out by nested PCR amplification of the 16S ribosomal RNA gene accompanied by DNA sequencing and analysis for verification. Results: A total of 10 tick species were identified as follows: Ornithodoros lahorensis (O. lahorensis (44.8%, Hyalomma dromedarii (15.6%, Dermacentor marginatus (13.5%, Hyalomma anatolicum (11.2%, Hyalomma asiaticum (5.7%, Hyalomma marginatum (4.9%, Rhipicephalus sanguineus (2.3%, Hyalomma detritum (1.0%, Dermacentor niveus (0.5% and Argas persicus (0.3%. The percentage distribution of Anaplasma/Ehrlichia was 55.5% (213 across 384 studied ticks. Conclusions: To the best of our knowledge, this is the first report of Anaplasma ovis infection in O. lahorensis in Iran. We also conjecture the prevalence of Ehrlichia spp. in Yazd Province based on sequencing results; also, it is suggested that O. lahorensis is a potential vector in the studied area. This survey highlights the importance of Argasidae family to verify and correlate their threat in causing anaplasmosis and other diseases in animals.

The Life and Death of Anaplasma

Centers for Disease Control (CDC) Podcasts

2012-03-30

Dr. Setu Vora, medical director of critical care and physician director of performance improvement at Backus Hospital in Norwich, Connecticut, reads his poem The Life and Death of Anaplasma and discusses the poem’s origins.  Created: 3/30/2012 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 4/2/2012.

Ixodes scapularis saliva mitigates inflammatory cytokine secretion during Anaplasma phagocytophilum stimulation of immune cells

Czech Academy of Sciences Publication Activity Database

Chen, G.; Severo, M. S.; Sohail, M.; Sakhon, O. S.; Wikel, S. K.; Kotsyfakis, Michalis; Pedra, J. H. F.

2012-01-01

Roč. 5, č. 1 (2012), s. 229 ISSN 1756-3305 Institutional support: RVO:60077344 Keywords : Tick * Ixodes scapularis * Saliva * Anaplasma phagocytophilum * Rickettsial agent Subject RIV: EC - Immunology Impact factor: 3.246, year: 2012 http://www.parasitesandvectors.com/content/5/1/229

The physical and functional interaction of NDRG2 with MSP58 in cells

International Nuclear Information System (INIS)

Zhang Jing; Liu Junye; Li Xia; Li Fuyang; Wang Lifeng; Zhang Jian; Liu Xinping; Shen Lan; Liu Na; Deng Yanchun; Yang Angang; Han Hua; Zhao Mujun; Yao Libo

2007-01-01

NDRG2, a member of N-Myc downstream regulated gene family, exerts the important functions in cell differentiation and tumor suppression. Although the ectopic expressed Ndrg2 inhibits the proliferation of tumor cells, its intracellular signal transduction pathway is hardly known. Here, we identified MSP58, a 58-kDa microspherule protein, as an interacting partner of human Ndrg2 by using yeast two-hybrid screening. The interaction was confirmed by glutathione S-transferase pull-down assay in vitro and by co-immune-precipitation assay in vivo. The forkhead associated domain of MSP58 is essential for its interaction with Ndrg2. Ndrg2 could co-localize with MSP58 in nuclear of HeLa cell during cell stress. Furthermore, the modulation of Ndrg2 level influences the cell cycle process together with MSP58. In conclusion, the findings offered a novel insight into the physiological roles of Ndrg2

Investigating the Adult Ixodid Tick Populations and Their Associated Anaplasma, Ehrlichia, and Rickettsia Bacteria at a Rocky Mountain Spotted Fever Hotspot in Western Tennessee.

Science.gov (United States)

Trout Fryxell, Rebecca T; Hendricks, Brain M; Pompo, Kimberly; Mays, Sarah E; Paulsen, Dave J; Operario, Darwin J; Houston, Allan E

2017-08-01

Ehrlichiosis and rickettsiosis are two common bacterial tick-borne diseases in the southeastern United States. Ehrlichiosis is caused by ehrlichiae transmitted by Amblyomma americanum and rickettsiosis is caused by rickettsiae transmitted by Amblyomma maculatum and Dermacentor variabilis. These ticks are common and have overlapping distributions in the region. The objective of this study was to identify Anaplasma, Ehrlichia, and Rickettsia species associated with questing ticks in a Rocky Mountain spotted fever (RMSF) hotspot, and identify habitats, time periods, and collection methods for collecting questing-infected ticks. Using vegetation drags and CO 2 -baited traps, ticks were collected six times (May-September 2012) from 100 sites (upland deciduous, bottomland deciduous, grassland, and coniferous habitats) in western Tennessee. Adult collections were screened for Anaplasma and Ehrlichia (simultaneous polymerase chain reaction [PCR]) and Rickettsia using genus-specific PCRs, and resulting positive amplicons were sequenced. Anaplasma and Ehrlichia were only identified within A. americanum (Ehrlichia ewingii, Ehrlichia chaffeensis, Panola Mountain Ehrlichia, and Anaplasma odocoilei sp. nov.); more Ehrlichia-infected A. americanum were collected at the end of June regardless of habitat and collection method. Rickettsia was identified in three tick species; "Candidatus Rickettsia amblyommii" from A. americanum, R. parkeri and R. andeanae from A. maculatum, and R. montanensis ( = montana) from D. variabilis. Overall, significantly more Rickettsia-infected ticks were identified as A. americanum and A. maculatum compared to D. variabilis; more infected-ticks were collected from sites May-July and with dragging. In this study, we report in the Tennessee RMSF hotspot the following: (1) Anaplasma and Ehrlichia are only found in A. americanum, (2) each tick species has its own Rickettsia species, (3) a majority of questing-infected ticks are collected May-July, (4) A

Middlesex Sampling Plant [MSP] annual site environmental report, calendar year 1988

International Nuclear Information System (INIS)

1989-04-01

The environmental monitoring program, which began in 1980, was continued in 1988 at the former Middlesex Sampling Plant (MSP) site, located in the Borough of Middlesex, New Jersey. The MSP site is part of the Formerly Utilized Site Remedial Action Program (FUSRAP), a Department of Energy (DOE) program to decontaminate or otherwise control sites where residual radioactive materials remain either from the early years of the nation's atomic energy program or from commercial operations causing conditions that Congress has mandated DOE to remedy. The environmental monitoring program is carried out by Bechtel National, Inc. (BNI), project management contractor for FUSRAP. The monitoring program at the MSP measures radon concentrations in air; external gamma radiation levels; and uranium and radium concentrations in surface water, groundwater, and sediment. To verify that the site is in compliance with the DOE radiation protection standard (100 mrem/yr) and to assess its potential effect on public health, the radiation dose was calculated for a hypothetical maximally exposed individual. Results of the 1988 monitoring show that the MSP is in compliance with applicable DOE radiation protection standards and with applicable requirements specified by New Jersey Department of Environmental Protection groundwater permits. 17 refs., 15 figs., 21 tabs

Detection of Ehrlichia spp., Anaplasma spp., Rickettsia spp., and Other Eubacteria in Ticks from the Thai-Myanmar Border and Vietnam

OpenAIRE

Parola, Philippe; Cornet, Jean-Paul; Sanogo, Yibayiri Osée; Miller, R. Scott; Thien, Huynh Van; Gonzalez, Jean-Paul; Raoult, Didier; Telford III, Sam R.; Wongsrichanalai, Chansuda

2003-01-01

A total of 650 ticks, including 13 species from five genera, were collected from animals, from people, or by flagging of the vegetation at sites on the Thai-Myanmar border and in Vietnam. They were tested by PCR to detect DNA of bacteria of the order Rickettsiales. Three Anaplasma spp. were detected in ticks collected in Thailand, including (i) Anaplasma sp. strain AnDa465, which was considered a genotype of Anaplasma platys (formerly Ehrlichia platys) and which was obtained from Dermacentor ...

Arnaldo Saraiva y las literaturas marginales y marginalizadas

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Carlos Nogueira

2014-04-01

Full Text Available This article focuses on Arnaldo Saraiva’s studies and teaching in the area of the socalled marginal or marginalized literatures of Portugal and Brazil, and on the unusually significant impact they have had on successive generations of teachers, cultural agents and literary scholars. RESUMEN: El presente trabajo versa sobre los estudios y la actividad docente de Arnaldo Saraiva en el área de las llamadas literaturas marginales o marginalizadas de Portugal y de Brasil, así como el impacto extrañamente significativo que aquellos han tenido en sucesivas generaciones de profesores, agentes culturales o estudiosos de la literatura.

Whole genome transcription profiling of Anaplasma phagocytophilum in human and tick host cells by tiling array analysis

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Chavez Adela

2008-07-01

Full Text Available Abstract Background Anaplasma phagocytophilum (Ap is an obligate intracellular bacterium and the agent of human granulocytic anaplasmosis, an emerging tick-borne disease. Ap alternately infects ticks and mammals and a variety of cell types within each. Understanding the biology behind such versatile cellular parasitism may be derived through the use of tiling microarrays to establish high resolution, genome-wide transcription profiles of the organism as it infects cell lines representative of its life cycle (tick; ISE6 and pathogenesis (human; HL-60 and HMEC-1. Results Detailed, host cell specific transcriptional behavior was revealed. There was extensive differential Ap gene transcription between the tick (ISE6 and the human (HL-60 and HMEC-1 cell lines, with far fewer differentially transcribed genes between the human cell lines, and all disproportionately represented by membrane or surface proteins. There were Ap genes exclusively transcribed in each cell line, apparent human- and tick-specific operons and paralogs, and anti-sense transcripts that suggest novel expression regulation processes. Seven virB2 paralogs (of the bacterial type IV secretion system showed human or tick cell dependent transcription. Previously unrecognized genes and coding sequences were identified, as were the expressed p44/msp2 (major surface proteins paralogs (of 114 total, through elevated signal produced to the unique hypervariable region of each – 2/114 in HL-60, 3/114 in HMEC-1, and none in ISE6. Conclusion Using these methods, whole genome transcription profiles can likely be generated for Ap, as well as other obligate intracellular organisms, in any host cells and for all stages of the cell infection process. Visual representation of comprehensive transcription data alongside an annotated map of the genome renders complex transcription into discernable patterns.

MGMT promoter methylation determined by HRM in comparison to MSP and pyrosequencing for predicting high-grade glioma response.

Science.gov (United States)

Switzeny, Olivier J; Christmann, Markus; Renovanz, Mirjam; Giese, Alf; Sommer, Clemens; Kaina, Bernd

2016-01-01

The DNA repair protein O(6)-methylguanine-DNA methyltransferase (MGMT) causes resistance of cancer cells to alkylating agents and, therefore, is a well-established predictive marker for high-grade gliomas that are routinely treated with alkylating drugs. Since MGMT is highly epigenetically regulated, the MGMT promoter methylation status is taken as an indicator of MGMT silencing, predicting the outcome of glioma therapy. MGMT promoter methylation is usually determined by methylation specific PCR (MSP), which is a labor intensive and error-prone method often used semi-quantitatively. Searching for alternatives, we used closed-tube high resolution melt (HRM) analysis, which is a quantitative method, and compared it with MSP and pyrosequencing regarding its predictive value. We analyzed glioblastoma cell lines with known MGMT activity and formalin-fixed samples from IDH1 wild-type high-grade glioma patients (WHO grade III/IV) treated with radiation and temozolomide by HRM, MSP, and pyrosequencing. The data were compared as to progression-free survival (PFS) and overall survival (OS) of patients exhibiting the methylated and unmethylated MGMT status. A promoter methylation cut-off level relevant for PFS and OS was determined. In a multivariate Cox regression model, methylation of MGMT promoter of high-grade gliomas analyzed by HRM, but not MSP, was found to be an independent predictive marker for OS. Univariate Kaplan-Meier analyses revealed for PFS and OS a significant and better discrimination between methylated and unmethylated tumors when quantitative HRM was used instead of MSP. Compared to MSP and pyrosequencing, the HRM method is simple, cost effective, highly accurate and fast. HRM is at least equivalent to pyrosequencing in quantifying the methylation level. It is superior in predicting PFS and OS of high-grade glioma patients compared to MSP and, therefore, can be recommended being used routinely for determination of the MGMT status of gliomas.

Development of duplex PCR for simultaneous detection of Theileria spp. and Anaplasma spp. in sheep and goats.

Science.gov (United States)

Cui, Yanyan; Zhang, Yan; Jian, Fuchun; Zhang, Longxian; Wang, Rongjun; Cao, Shuxuan; Wang, Xiaoxing; Yan, Yaqun; Ning, Changshen

2017-05-01

Theileria spp. and Anaplasma spp., which are important tick-borne pathogens (TBPs), impact the health of humans and animals in tropical and subtropical areas. Theileria and Anaplasma co-infections are common in sheep and goats. Following alignment of the relevant DNA sequences, two primer sets were designed to specifically target the Theileria spp. 18S rRNA and Anaplasma spp. 16S rRNA gene sequences. Genomic DNA from the two genera was serially diluted tenfold for testing the sensitivities of detection of the primer sets. The specificities of the primer sets were confirmed when DNA from Anaplasma and Theileria (positive controls), other related hematoparasites (negative controls) and ddH 2 O were used as templates. Fifty field samples were also used to evaluate the utility of single PCR and duplex PCR assays, and the detection results were compared with those of the PCR methods previously published. An optimized duplex PCR assay was established from the two primer sets based on the relevant genes from the two TBPs, and this assay generated products of 298-bp (Theileria spp.) and 139-bp (Anaplasma spp.). The detection limit of the assay was 29.4 × 10 -3  ng per μl, and there was no cross-reaction with the DNA from other hematoparasites. The results showed that the newly developed duplex PCR assay had an efficiency of detection (P > 0.05) similar to other published PCR methods. In this study, a duplex PCR assay was developed that can simultaneously identify Theileria spp. and Anaplasma spp. in sheep and goats. This duplex PCR is a potentially valuable assay for epidemiological studies of TBPs in that it can detect cases of mixed infections of the pathogens. Copyright © 2017 Elsevier Inc. All rights reserved.

Satisfactory safety and immunogenicity of MSP3 malaria vaccine candidate in Tanzanian children aged 12–24 months

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Segeja Method D

2009-07-01

Full Text Available Abstract Background Development and deployment of an effective malaria vaccine would complement existing malaria control measures. A blood stage malaria vaccine candidate, Merozoite Surface Protein-3 (MSP3, produced as a long synthetic peptide, has been shown to be safe in non-immune and semi-immune adults. A phase Ib dose-escalating study was conducted to assess the vaccine's safety and immunogenicity in children aged 12 to 24 months in Korogwe, Tanzania (ClinicalTrials.gov number: NCT00469651. Methods This was a double-blind, randomized, controlled, dose escalation phase Ib trial, in which children were given one of two different doses of the MSP3 antigen (15 μg or 30 μg or a control vaccine (Engerix B. Children were randomly allocated either to the MSP3 candidate malaria vaccine or the control vaccine administered at a schedule of 0, 1, and 2 months. Immunization with lower and higher doses was staggered for safety reasons starting with the lower dose. The primary endpoint was safety and reactogenicity within 28 days post-vaccination. Blood samples were obtained at different time points to measure immunological responses. Results are presented up to 84 days post-vaccination. Results A total of 45 children were enrolled, 15 in each of the two MSP3 dose groups and 15 in the Engerix B group. There were no important differences in reactogenicity between the two MSP3 groups and Engerix B. Grade 3 adverse events were infrequent; only five were detected throughout the study, all of which were transient and resolved without sequelae. No serious adverse event reported was considered to be related to MSP3 vaccine. Both MSP3 dose regimens elicited strong cytophilic IgG responses (subclasses IgG1 and IgG3, the isotypes involved in the monocyte-dependant mechanism of Plasmodium falciparum parasite-killing. The titers reached are similar to those from African adults having reached a state of premunition. Furthermore, vaccination induced seroconversion in

MSP-HTPrimer: a high-throughput primer design tool to improve assay design for DNA methylation analysis in epigenetics.

Science.gov (United States)

Pandey, Ram Vinay; Pulverer, Walter; Kallmeyer, Rainer; Beikircher, Gabriel; Pabinger, Stephan; Kriegner, Albert; Weinhäusel, Andreas

2016-01-01

Bisulfite (BS) conversion-based and methylation-sensitive restriction enzyme (MSRE)-based PCR methods have been the most commonly used techniques for locus-specific DNA methylation analysis. However, both methods have advantages and limitations. Thus, an integrated approach would be extremely useful to quantify the DNA methylation status successfully with great sensitivity and specificity. Designing specific and optimized primers for target regions is the most critical and challenging step in obtaining the adequate DNA methylation results using PCR-based methods. Currently, no integrated, optimized, and high-throughput methylation-specific primer design software methods are available for both BS- and MSRE-based methods. Therefore an integrated, powerful, and easy-to-use methylation-specific primer design pipeline with great accuracy and success rate will be very useful. We have developed a new web-based pipeline, called MSP-HTPrimer, to design primers pairs for MSP, BSP, pyrosequencing, COBRA, and MSRE assays on both genomic strands. First, our pipeline converts all target sequences into bisulfite-treated templates for both forward and reverse strand and designs all possible primer pairs, followed by filtering for single nucleotide polymorphisms (SNPs) and known repeat regions. Next, each primer pairs are annotated with the upstream and downstream RefSeq genes, CpG island, and cut sites (for COBRA and MSRE). Finally, MSP-HTPrimer selects specific primers from both strands based on custom and user-defined hierarchical selection criteria. MSP-HTPrimer produces a primer pair summary output table in TXT and HTML format for display and UCSC custom tracks for resulting primer pairs in GTF format. MSP-HTPrimer is an integrated, web-based, and high-throughput pipeline and has no limitation on the number and size of target sequences and designs MSP, BSP, pyrosequencing, COBRA, and MSRE assays. It is the only pipeline, which automatically designs primers on both genomic

Molecular detection of tick-borne pathogens in cattle from Southwestern Ethiopia.

Directory of Open Access Journals (Sweden)

Zerihun Hailemariam

Full Text Available Tick-borne diseases (TBDs cause significant losses among livestock and impact the livelihoods of resource-poor farming communities worldwide. In Ethiopia, detailed studies on the epidemiology of tick-borne pathogens (TBPs in cattle using sensitive molecular detection methods are scarce. The objective of this study was to determine the prevalence and species composition of bovine TBPs of veterinary significance in local cattle populations. A comprehensive cross-sectional epidemiological study was conducted in cattle populations of Illubabor zone in Southwestern Ethiopia from June to August 2013. For this purpose, blood samples were collected from 392 cattle. A combination of polymerase chain reaction (PCR and a Reverse Line Blot (RLB hybridization assay was employed for the detection of TBPs in these samples. The PCR/RLB results of the 392 blood samples indicated a high overall prevalence of 96.9% for TBPs, including Theileria mutans (66.1%, Theileria orientalis (51.8%, Anaplasma sp. Omatjenne (25.5%, Anaplasma marginale (14.5%, Babesia bigemina (14.0% and Theileria velifera (13.0% and minor occurrences of Ehrlichia ruminantium (0.5% and Ehrlichia minasensis (0.26%. Moreover, three novel Anaplasma genotypes were detected in bovine blood samples. A phylogenetic analysis revealed that they most likely represent three, but at least two, new species. The prevalence of the three novel Anaplasma species, preliminary designated as Anaplasma sp. Hadesa, Anaplasma sp. Saso and Anaplasma sp. Dedessa, was 12.5%, 14.3% and 5.6%, respectively. Overall, a total of 227 cattle (57.9% were found to be co-infected with two or more TBPs simultaneously and 86 different species combinations were observed. The findings show a very high burden of infection of cattle with TBPs in Ethiopia. The high frequency of co-infections suggests that clinical manifestations might be complex. Further research is required to determine the pathogenicity, host cell types and vector of

Detection of Anaplasma sp. phylogenetically related to A. phagocytophilum in a free-living bird in Brazil

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Anna Claudia Baumel Mongruel

2017-08-01

Full Text Available Abstract Wild animals play an important role in carrying vectors that may potentially transmit pathogens. Several reports highlighted the participation of wild animals on the Anaplasma phagocytophilum cycle, including as hosts of the agent. The aim of this study was to report the molecular detection of an agent phylogenetically related to A. phagocytophilum isolated from a wild bird in the Midwest of the state of Paraná, Brazil. Fifteen blood samples were collected from eleven different bird species in the Guarapuava region. One sample collected from a Penelope obscura bird was positive in nested PCR targeting the 16S rRNA gene of Anaplasma spp. The phylogenetic tree based on the Maximum Likelihood analysis showed that the sequence obtained was placed in the same clade with A. phagocytophilum isolated from domestic cats in Brazil. The present study reports the first molecular detection of a phylogenetically related A. phagocytophilum bacterium in a bird from Paraná State.

Genome sequencing and annotation of Acinetobacter guillouiae strain MSP 4-18

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Nitin Kumar Singh

2014-12-01

Full Text Available The genus Acinetobacter consists of 31 validly published species ubiquitously distributed in nature and primarily associated with nosocomial infection. We report the 4.8 Mb genome of Acinetobacter guillouiae MSP 4-18, isolated from a mangrove soil sample from Parangipettai (11°30′N, 79°47′E, Tamil Nadu, India. The draft genome of A. guillouiae MSP 4-18 has a G + C content of 38.0% and includes 3 rRNA genes (5S, 23S, 16S and 69 aminoacyl-tRNA synthetase genes.

Region of interest methylation analysis: a comparison of MSP with MS-HRM and direct BSP.

Science.gov (United States)

Akika, Reem; Awada, Zainab; Mogharbil, Nahed; Zgheib, Nathalie K

2017-07-01

The aim of this study was to compare and contrast three DNA methylation methods of a specific region of interest (ROI): methylation-specific PCR (MSP), methylation-sensitive high resolution melting (MS-HRM) and direct bisulfite sequencing (BSP). The methylation of a CpG area in the promoter region of Estrogen receptor alpha (ESR1) was evaluated by these three methods with samples and standards of different methylation percentages. MSP data were neither reproducible nor sensitive, and the assay was not specific due to non-specific binding of primers. MS-HRM was highly reproducible and a step forward into categorizing the methylation status of the samples as percent ranges. Direct BSP was the most informative method regarding methylation percentage of each CpG site. Though not perfect, it was reproducible and sensitive. We recommend the use of either method depending on the research question and target amplicon, and provided that the designed primers and expected amplicons are within recommendations. If the research question targets a limited number of CpG sites and simple yes/no results are enough, MSP may be attempted. For short amplicons that are crowded with CpG sites and of single melting domain, MS-HRM may be the method of choice though it only indicates the overall methylation percentage of the entire amplicon. Although the assay is highly reproducible, being semi-quantitative makes it of lesser interest to study ROI methylation of samples with little methylation differences. Direct BSP is a step forward as it gives information about the methylation percentage at each CpG site.

Multiple-specimen absolute paleointensity determination with the MSP-DSC protocol: Advantages and drawbacks.

Science.gov (United States)

Camps, P.; Fanjat, G.; Poidras, T.; Carvallo, C.; Nicol, P.

2012-04-01

The MSP-DSC protocol (Dekkers & Bohnel, 2006, EPSL; Fabian & Leonhardt, 2010, EPSL) is a recent development in the methodology for documenting the intensity of the ancient Earth magnetic field. Applicable both on rocks or archaeological artifacts it allows us to use samples that until now were not measured because their magnetic properties do not meet selection criteria required by conventional methods. However, this new experimental protocol requires that samples be heated and cooled under a field parallel to its natural remanent magnetization (NRM). Currently, standard paleointensity furnaces do not match precisely this constraint. Yet, such new measurement protocol seems very promising since it would possibly double the number of available data. We are developing in Montpellier (France), a very fast-heating oven with infrared dedicated to this protocol. Two key points determine its characteristics. The first is to heat uniformly a rock sample of a 10-cc-standard volume as fast as possible. The second is to apply to the sample during the heating (and the cooling) a precise magnetic induction field, perfectly controlled in 3D. We tested and calibrated a preliminary version of this oven along with the MSP-DSC protocol with 3 historical lava flows, 2 from Reunion Island (erupted in 2002 and 2007) and one from Etna (erupted in 1983). These lava flows were selected because they have different magnetic behaviors. Reunion 2002 is rather SD-PSD-like, while Reunion 2007 is PSD-MD-like, and Etna 1983 is MD-like. The paleointensity determinations obtained with the original protocol of Dekkers and Bohnel (2006, EPSL) are within +- 1 μT of the known field for the three lava flows. The same precision is obtained when we applied the fraction correction (MSP-FC protocol). However, we systematically observed a loss in the linearity of the MSP-FC plots. In addition, like Muxworthy and Taylor (2011, GJI), we found that the Domain State Correction is difficult to apply since alpha

Effectiveness of enrofloxacin for the treatment of experimentally-induced bovine anaplasmosis Eficácia da enrofloxacina no tratamento da anaplasmose bovina experimental

Directory of Open Access Journals (Sweden)

Elias Jorge Facury-Filho

2012-03-01

Full Text Available Four groups of six Holstein calves were inoculated with 3.6 × 10(7 erythrocytes parasitized with Anaplasma marginale. The criteria for treatment of calves were increasing A. marginale rickettsemia and 30% reduction of baseline packed cell volume (PCV of each animal. Group 1 (G1 received 7.5 mg.kg-1 of enrofloxacin in a single dose; Group 2 (G2 7.5 mg.kg-1 of enrofloxacin twice every three days; Group 3 (G3 20 mg.kg-1 of long-acting oxytetracycline in a single dose; and Group 4 (G4 a single dose of PBS. Physical examinations, blood smears and PCV were performed daily. On day treatment, G1, G2 and G3 animals had a mean rickettsemia of 17, 23 and 12%, respectively. At 2 days after treatment (DAT G1 and G2 animals showed a significant reduction of rickettsemia (p Quatro grupos de seis bezerros da raça Holandesa foram inoculados com 3,6 × 10(7 eritrócitos parasitados com Anaplasma marginale. Os critérios para o tratamento dos bezerros foram aumento da rickettsemia do A. marginale e redução de 30% do valor basal de volume globular (VG de cada animal. O Grupo 1 (G1 recebeu 7,5 mg.kg-1 de enrofloxacina em dose única; o Grupo 2 (G2, 7,5 mg.kg-1 de enrofloxacina duas vezes a cada três dias; o Grupo 3 (G3, 20 mg.kg-1 de oxitetraciclina de longa ação em dose única; e o Grupo 4 (G4 uma única dose de PBS. Exames físicos, esfregaço sanguíneo e VG foram realizadas diariamente. No dia do tratamento, os animais G1, G2 e G3 apresentaram rickettsemia média de 17, 23 e 12%, respectivamente. Nos primeiros 2 dias após o tratamento (DAT os animais do G1 e G2 mostraram uma redução significativa de rickettsemia (p < 0,05 em relação ao G3. Animais do G3 tiveram altas taxas de rickettsemia nos 2 DAT e uma diminuição lenta até à estabilização em 9 de DAT. O VG médio no G1 e G2 aumentou e estabilizou após 7 e 8 DAT, respectivamente. A estabilização do VG do G3 foi aos 13 DAT. A enrofloxacina e a oxitetraciclina foram efetivas no tratamento da

Occurrence of Hepatozoon canis (Adeleorina: Hepatozoidae) and Anaplasma spp. (Rickettsiales: Anaplasmataceae) in black-backed jackals (Canis mesomelas) in South Africa.

Science.gov (United States)

Penzhorn, Barend L; Netherlands, Edward C; Cook, Courtney A; Smit, Nico J; Vorster, Ilse; Harrison-White, Robert F; Oosthuizen, Marinda C

2018-03-20

Domestic dogs are not native to sub-Saharan Africa, which may account for their susceptibility to Babesia rossi, of which endemic black-backed jackals (Canis mesomelas) are natural reservoirs. There is virtually no information on the occurrence of potentially pathogenic haemogregarines (e.g. Hepatozoon canis) or even rickettsial bacteria (e.g. Ehrlichia spp. and Anaplasma spp.) in indigenous canids in sub-Saharan Africa. Such organisms could pose a risk to domestic dogs, as well as to populations of endangered indigenous canid species. Genomic DNA extracted from blood samples taken from 126 free-ranging and 16 captive black-backed jackals was subjected to reverse line blot (RLB) hybridization assay; 82 (57.8%) specimens reacted only with the Ehrlichia/Anaplasma genera-specific probe. Full-length bacterial 16S rRNA gene of five of these specimens was cloned and the recombinants sequenced. The ten 16S rDNA sequences obtained were most closely related, with approximately 99% identity, to Anaplasma sp. South African Dog, various uncultured Anaplasma spp., as well as various Anaplasma phagocytophilum genotypes. Ninety-one specimens were screened for haemogregarines through PCR amplification using the 18S rRNA gene; 20 (21.9%) specimens reacted positively, of which 14 (15.4%) were confirmed positive for Hepatozoon genotypes from within H. canis. Two (2.2%) specimens were found positive for two different Hepatozoon genotypes. Sequence analyses confirmed the presence of 16S rDNA sequences closely related to A. phagocytophilum and Anaplasma sp. South African Dog as well as two H. canis genotypes in both free-ranging and captive black-backed jackals. Distinguishing between closely related lineages may provide insight into differences in pathogenicity and virulence of various Anaplasma and H. canis genotypes. By building up a more comprehensive understanding of the range and diversity of the bacteria and eukaryotic organisms (piroplasms and haemogregarines) in the blood of

Citrate synthase gene sequence: a new tool for phylogenetic analysis and identification of Ehrlichia.

Science.gov (United States)

Inokuma, H; Brouqui, P; Drancourt, M; Raoult, D

2001-09-01

The sequence of the citrate synthase gene (gltA) of 13 ehrlichial species (Ehrlichia chaffeensis, Ehrlichia canis, Ehrlichia muris, an Ehrlichia species recently detected from Ixodes ovatus, Cowdria ruminantium, Ehrlichia phagocytophila, Ehrlichia equi, the human granulocytic ehrlichiosis [HGE] agent, Anaplasma marginale, Anaplasma centrale, Ehrlichia sennetsu, Ehrlichia risticii, and Neorickettsia helminthoeca) have been determined by degenerate PCR and the Genome Walker method. The ehrlichial gltA genes are 1,197 bp (E. sennetsu and E. risticii) to 1,254 bp (A. marginale and A. centrale) long, and GC contents of the gene vary from 30.5% (Ehrlichia sp. detected from I. ovatus) to 51.0% (A. centrale). The percent identities of the gltA nucleotide sequences among ehrlichial species were 49.7% (E. risticii versus A. centrale) to 99.8% (HGE agent versus E. equi). The percent identities of deduced amino acid sequences were 44.4% (E. sennetsu versus E. muris) to 99.5% (HGE agent versus E. equi), whereas the homology range of 16S rRNA genes was 83.5% (E. risticii versus the Ehrlichia sp. detected from I. ovatus) to 99.9% (HGE agent, E. equi, and E. phagocytophila). The architecture of the phylogenetic trees constructed by gltA nucleotide sequences or amino acid sequences was similar to that derived from the 16S rRNA gene sequences but showed more-significant bootstrap values. Based upon the alignment analysis of the ehrlichial gltA sequences, two sets of primers were designed to amplify tick-borne Ehrlichia and Neorickettsia genogroup Ehrlichia (N. helminthoeca, E. sennetsu, and E. risticii), respectively. Tick-borne Ehrlichia species were specifically identified by restriction fragment length polymorphism (RFLP) patterns of AcsI and XhoI with the exception of E. muris and the very closely related ehrlichia derived from I. ovatus for which sequence analysis of the PCR product is needed. Similarly, Neorickettsia genogroup Ehrlichia species were specifically identified by

Antibodies against PfEMP1, RIFIN, MSP3 and GLURP are acquired during controlled Plasmodium falciparum malaria infections in naïve volunteers

DEFF Research Database (Denmark)

Turner, Louise; Wang, Christian W; Lavstsen, Thomas

2011-01-01

Antibodies to polymorphic antigens expressed during the parasites erythrocytic stages are important mediators of protective immunity against P. falciparum malaria. Therefore, polymorphic blood stage antigens like MSP3, EBA-175 and GLURP and variant surface antigens PfEMP1 and RIFIN are considered...

Usos marginales de los marcadores del discurso. Su efecto en la comprensión lectora en español como lengua extranjera

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María Fernanda Pelayo

2003-04-01

Full Text Available En este trabajo abordamos lo que hemos denominado usos marginales de los marcadores del discurso y cómo afectan a la comprensión lectora en español como lengua extranjera. Tras un análisis y definición de estos usos no ortodoxos de los marcadores del discurso, realizamos una clasificación preliminar de los mismos. Finalmente, diseñamos y llevamos a cabo un estudio empírico acerca de la manera en que los usos marginales de los marcadores del discurso podrían afectar a la comprensión lectora de textos en español como lengua extranjera. Losresultados obtenidos en este estudio experimental muestran que existe una tendencia bastante generalizada que nos hace aventurarnos a concluir que efectivamente los lectores de español como lengua extranjera muestran tenerproblemas para comprender textos en los que se han empleado usos marginales de los marcadores del discurso. Por otra parte, el uso ortodoxo de los marcadores del discurso parece favorecer la comprensión adecuada del texto.

Molecular detection of Anaplasma bovis, Ehrlichia canis and Hepatozoon felis in cats from Luanda, Angola.

Science.gov (United States)

Oliveira, Ana Cristina; Luz, Maria Francisca; Granada, Sara; Vilhena, Hugo; Nachum-Biala, Yaarit; Lopes, Ana Patrícia; Cardoso, Luís; Baneth, Gad

2018-03-20

Molecular identification of tick-borne pathogen infection in cats from Africa is scarce. The presence of bacterial (Anaplasma and Ehrlichia) and protozoal (Babesia and Hepatozoon) agents was investigated in blood samples from 102 domestic cats from Luanda, Angola, by polymerase chain reaction and DNA sequencing. Three cats (2.9%) were found infected with Ehrlichia canis, three (2.9%) with Hepatozoon felis and one (1.0%) with Anaplasma bovis. The prevalence of infections with one single agent was 4.9%, and that of infection with two agents (i.e. E. canis and H. felis) was 1.0%. In total, six cats (5.9%) were found infected with at least one of the detected tick-borne agents. This is the first report of A. bovis, E. canis and H. felis in cats from Angola. To the best of our knowledge, A. bovis is also being reported for the first time in domestic cats outside of Japan. Cats are at a low to moderate risk of being infected with tick-borne agents in Luanda.

Bovine anaplasmosis with emphasis on immune responses and protection

International Nuclear Information System (INIS)

Ristic, M.

1980-01-01

Anaplasmosis is an infectious and transmissible disease manifested by progressive anaemia and the appearance of other characteristic disease symptoms. It is a world-wide tick-borne disease of cattle and some wild ruminants caused by the rickettsia Anaplasma marginale. By drawing on information obtained from studies of plasmodial cell cultures, a method has recently been developed for short-term in vitro cultivation of A. marginale. An attenuated Anaplasma organism capable of growth in both ovine and bovine erythrocytes was used to demonstrate that the in vitro system provided the necessary requirements for active transfer of the organism from cell to cell. Organismal antigens are found in the erythrocytes of infected animals, whereas soluble antigens are derived from their erythrocytes and serum. Serums from convalescing animals interact with these antigens in agglutination, complement fixation, fluorescent antibody and precipitation tests. Passive transfer of sera from immune to susceptible cattle, however, does not seem to confer protection against the infection and development of the disease. Studies that employed various tests for measuring cell-mediated immune (CMI) responses (leukocyte migration inhibition, blast transformation and cytotoxicity), in association with information collected simultaneously on antibody activity, have shown that both humoral and cellular immune responses are needed for the development of protective immunity in anaplasmosis. It was further shown that an active replication of Anaplasma is essential for induction of these two types of immune responses. Consequently, live virulent and attenuated immunogens fulfil requirements for induction of protective immunity. With the virulent agent, however, development of protective immunity is preceded by induction of auto-immune responses apparently associated with pathogenesis of anaemia in anaplasmosis. Inactivated immunogens derived from blood of infected cattle and used in combination with

Detection of Ehrlichia spp., Anaplasma spp., Rickettsia spp., and other eubacteria in ticks from the Thai-Myanmar border and Vietnam.

Science.gov (United States)

Parola, Philippe; Cornet, Jean-Paul; Sanogo, Yibayiri Osée; Miller, R Scott; Thien, Huynh Van; Gonzalez, Jean-Paul; Raoult, Didier; Telford III, Sam R; Wongsrichanalai, Chansuda

2003-04-01

A total of 650 ticks, including 13 species from five genera, were collected from animals, from people, or by flagging of the vegetation at sites on the Thai-Myanmar border and in Vietnam. They were tested by PCR to detect DNA of bacteria of the order RICKETTSIALES: Three Anaplasma spp. were detected in ticks collected in Thailand, including (i) Anaplasma sp. strain AnDa465, which was considered a genotype of Anaplasma platys (formerly Ehrlichia platys) and which was obtained from Dermacentor auratus ticks collected from dogs; (ii) Anaplasma sp. strain AnAj360, which was obtained from Amblyomma javanense ticks collected on a pangolin; and (iii) Anaplasma sp. strain AnHl446, which was closely related to Anaplasma bovis and which was detected in Haemaphysalis lagrangei ticks collected from a bear. Three Ehrlichia spp. were identified, including (i) Ehrlichia sp. strain EBm52, which was obtained from Boophilus microplus ticks collected from cattle from Thailand; (ii) Ehrlichia sp. strain EHh324, which was closely related to Ehrlichia chaffeensis and which was detected in Haemaphysalis hystricis ticks collected from wild pigs in Vietnam; and (iii) Ehrlichia sp. strain EHh317, which was closely related to Ehrlichia sp. strain EBm52 and which was also detected in H. hystricis ticks collected from wild pigs in Vietnam. Two Rickettsia spp. were detected in Thailand, including (i) Rickettsia sp. strain RDla420, which was detected in Dermacentor auratus ticks collected from a bear, and (ii) Rickettsia sp. strain RDla440, which was identified from two pools of Dermacentor larvae collected from a wild pig nest. Finally, two bacteria named Eubacterium sp. strain Hw124 and Eubacterium sp. strain Hw191 were identified in Haemaphysalis wellingtoni ticks collected from chicken in Thailand; these strains could belong to a new group of bacteria.

Use of multiplex polymerase chain reaction-based assay to conduct epidemiological studies on bovine hemoparasites in Mexico.

Science.gov (United States)

Figueroa, J V; Alvarez, J A; Ramos, J A; Vega, C A; Buening, G M

1993-01-01

A study was conducted to test the applicability of a Polymerase Chain Reaction (PCR)-based approach for the simultaneous detection of the bovine hemoparasites Babesia bigemina, B. bovis and Anaplasma marginale. Bovine blood samples from cattle ranches of a previously determined enzootic zone in the Yucatan Peninsula of Mexico, were collected from peripheral blood and processed for PCR analysis. Blood samples were subjected to DNA amplification by placing an aliquot in a reaction tube containing oligonucleotide primers specific for DNA of each hemoparasite species. The PCR products were detected by Dot-Blot nucleic acid hybridization utilizing nonradioactive, species-specific, digoxigenin PCR-labeled DNA probes. Four hundred twenty one field samples analyzed by the multiplex PCR-DNA probe assay showed 66.7%, 60.1% and 59.6% prevalence rates for B. bigemina, B. bovis and A. marginale, respectively. The multiplex PCR analysis showed that animals with single, double or triple infection could be detected with the parasite specific DNA probes. The procedure is proposed as a valuable tool for the epidemiological analysis in regions where the hemoparasite species are concurrently infecting cattle.

El impuesto al valor agregado en los contratos de campos marginales

OpenAIRE

Niquinga Salazar, Oscar Javier

2007-01-01

Esta investigación dirigida por los métodos de investigación sistemático, exegético, y teleológico, pretende analizar los contratos petroleros de Campos Marginales utilizados por la legislación ecuatoriana y específicamente, el papel de Impuesto al Valor Agregado en la producción de la Curva Base; ésta, se convierte en una figura contractual innovadora que ha producido una interesante discusión a nivel nacional; sin embargo, la propuesta de este trabajo va encaminado a encontra...

The evolution and diversity of a low complexity vaccine candidate, merozoite surface protein 9 (MSP-9), in Plasmodium vivax and closely related species.

Science.gov (United States)

Chenet, Stella M; Pacheco, M Andreína; Bacon, David J; Collins, William E; Barnwell, John W; Escalante, Ananias A

2013-12-01

The merozoite surface protein-9 (MSP-9) has been considered a target for an anti-malarial vaccine since it is one of many proteins involved in the erythrocyte invasion, a critical step in the parasite life cycle. Orthologs encoding this antigen have been found in all known species of Plasmodium parasitic to primates. In order to characterize and investigate the extent and maintenance of MSP-9 genetic diversity, we analyzed DNA sequences of the following malaria parasite species: Plasmodium falciparum, Plasmodium reichenowi, Plasmodium chabaudi, Plasmodium yoelii, Plasmodium berghei, Plasmodium coatneyi, Plasmodium gonderi, Plasmodium knowlesi, Plasmodium inui, Plasmodium simiovale, Plasmodium fieldi, Plasmodium cynomolgi and Plasmodium vivax and evaluated the signature of natural selection in all MSP-9 orthologs. Our findings suggest that the gene encoding MSP-9 is under purifying selection in P. vivax and closely related species. We further explored how selection affected different regions of MSP-9 by comparing the polymorphisms in P. vivax and P. falciparum, and found contrasting patterns between these two species that suggest differences in functional constraints. This observation implies that the MSP-9 orthologs in human parasites may interact differently with the host immune response. Thus, studies carried out in one species cannot be directly translated into the other. Copyright © 2013 Elsevier B.V. All rights reserved.

Functional and Immunological Relevance of Anaplasma marginale Major Surface Protein 1a Sequence and Structural Analysis

Czech Academy of Sciences Publication Activity Database

Cabezas Cruz, Alejandro; Passos, L.M.F.; Lis, K.; Kenneil, R.; Valdés, James J.; Ferrolho, J.; Tonk, Miray; Pohl, A.E.; Grubhoffer, Libor; Zweygarth, E.; Shkap, V.; Ribeiro, M.F.B.; Estrada-Pena, A.; Kocan, K.M.; de la Fuente, J.

2013-01-01

Roč. 8, č. 6 (2013), e65243 E-ISSN 1932-6203 R&D Projects: GA MŠk(CZ) EE2.3.30.0032 EU Projects: European Commission(XE) 238511 - POSTICK Institutional support: RVO:60077344 Keywords : CD4(+) T-lymphocytes * B-cell epitopes * salivary glands Subject RIV: EC - Immunology Impact factor: 3.534, year: 2013

Análisis de la dinámica relacional generada por las websites de Internet en el desarrollo de los turismos marginales

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José Mª Prat

2012-01-01

Full Text Available El conocimiento de los actores turísticos del territorio, y de sus relaciones, es un elemento básico para la gestión activa de cualquier destino turístico. Como que, gracias a las nuevas tecnologías de la información y a la popularizacióndel uso de Internet, se han creado una gran cantidad de relaciones sociales gracias a las websites, interrelacionadas, que han permitido una rápida y amplia difusión promocional de aquellos destinos que ofrecen unas alternativas turísticas más marginales, como respuesta a los nuevos planteamientos post-fordistas del turismo, generándose una dinámica relacional que se ha beneficiado del rápido desarrollo de las TIC's. En este trabajo se han analizado dichas relaciones entre websites, en el caso del desarrollo de dos turismos marginales, el naturista y el swinger, en Cap d'Agde, en el sur de Francia, comprobándose la existencia de una relación directa entre la dinámica relacional del territorio y elgrado de desarrollo de los turismos marginales.

Incidencia de los Hábitos Alimenticios en los Niveles de Rendimiento Académico Escolar en las Instituciones Educativas Secundarias Urbano Marginales de Juliaca

OpenAIRE

Aguilar Roque, Gaylee Yemira

2015-01-01

El presente trabajo de investigación denominado “INCIDENCIA DE LOS HÁBITOS ALIMENTICIOS EN LOS NIVELES DE RENDIMIENTO ACADÉMICO ESCOLAR EN LAS INSTITUCIONES EDUCATIVAS SECUNDARIAS URBANO - MARGINALES DE JULIACA”, cuyo objetivo general es establecer la importancia del consumo de cereales andinos y productos procesados en los niveles de rendimiento académico de los estudiantes de las I.E.S. de las zonas urbano – marginales de la ciudad de Juliaca. Este estudio nos permite establec...

NNDSS - Table II. Ehrlichiosis and Anaplasmosis, Anaplasma phagocytophilum infection to Ehrlichia chaffeensis infection

Data.gov (United States)

U.S. Department of Health & Human Services — NNDSS - Table II. Ehrlichiosis and Anaplasmosis, Anaplasma phagocytophilum infection to Ehrlichia chaffeensis infection - 2018. In this Table, provisional cases of...

Frequency of Anaplasma marginale (Theiler 1910 and Babesia sp in mestizo bovine Zebu, in the Municipality of Ixiamas county Abel Iturralde Department of The La Paz, Bolivia

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Mercado Alvaro

2011-08-01

Full Text Available The present work one carries out in the Municipality of Ixiamas County Abel Iturralde of the department of Peace-Bolivia, between May and August of 2010. The objective was to determine the presence of Anaplama sp and Babesia sp, through sanguine smear, they took 160 samples of blood, 40 bovine of the four areas they have been evaluated. The results indicate the presence of Anaplasma sp 6,90%, 6,20% in females and 9,70% in males, this represents 2,50% for the area TO, 5,00% area B, 17,50% area C and 2,50% area D. According to the category 12,50% in calves, 0,00% in torillos, 4,80% in bulls, 6,20% in cows and 14,30% in heifers. The presence of Babesia sp 3,13%, with relationship to the sex 3,10% in females and 3,22% in males, being observed 2,50% in the area TO, 5,00% in the area B, 5,00% in the area C and 0,00% in the area D, according to the category 0,00% in calves, torillos 4,80% bulls, 2,73% vacates and 7,14% in heifers.

Expression and Evaluation of Recombinant Plasmodium knowlesi Merozoite Surface Protein-3 (MSP-3 for Detection of Human Malaria.

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Jeremy Ryan De Silva

Full Text Available Malaria remains a major health threat in many parts of the globe and causes high mortality and morbidity with 214 million cases of malaria occurring globally in 2015. Recent studies have outlined potential diagnostic markers and vaccine candidates one of which is the merozoite surface protein (MSP-3. In this study, novel recombinant Plasmodium knowlesi MSP-3 was cloned, expressed and purified in an Escherichia coli system. Subsequently, the recombinant protein was evaluated for its sensitivity and specificity. The recombinant pkMSP-3 protein reacted with sera from patients with P. knowlesi infection in both Western blot (61% and ELISA (100%. Specificity-wise, pkMSP-3 did not react with healthy donor sera in either assay and only reacted with a few non-malarial parasitic patient sera in the ELISA assay (3 of 49. In conclusion, sensitivity and specificity of pkMSP-3 was found to be high in the ELISA and Western Blot assay and thus utilising both assays in tandem would provide the best sero-diagnostic result for P. knowlesi infection.

Natural antibody responses to Plasmodium falciparum MSP3 and GLURP(R0) antigens are associated with low parasite densities in malaria patients living in the Central Region of Ghana

DEFF Research Database (Denmark)

Amoah, L. E.; Nuvor, S. V.; Obboh, E. K.

2017-01-01

Background: Plasmodium falciparum genetic diversity and multiplicity of infection (MOI) are parasite features that have been suggested to influence the acquisition of protective immunity against malaria. This study sought to assess the relationship between MOI and parasite density (PD) in malaria...... and adults diagnosed with uncomplicated P. falciparum malaria. Microscopy was used to estimate P. falciparum parasite density and polymerase chain reaction (PCR) amplification of the polymorphic regions of msp1 (PF3D7-0930300) and msp2 (PF3D7-0206800) was used for parasite genotyping and MOI determination....... The geometric mean (GM) for MOI determined by both msp1 and msp2 genotyping was 1.3 for the entire population and was generally higher in children than in adults. Seropositivity was estimated at 67 and 63% for GLURP(R0) and MSP3 antibodies, respectively, and antibody titers were negatively correlated...

Internetová reklama pro MSP

OpenAIRE

Molnár, Peter

2007-01-01

Cielom práce je poukázať na hlavné formy internetovej reklamy, ktoré sa používajú pri reklamných kampaniach. Dalšími cielmi je poukázať na využitie internetovej reklamy z pohladu MSP ako aj návrh reklamnej kampane za použitia internetových foriem reklamy. The aim of this work is to point main forms of internet advertising, used by advertising campaign. The other aims are to point on usage of internet advertising in SME and layout of advertising campaign using internet forms of advertising....

Molecular detection and identification of hemoparasites in pampas deer (Ozotoceros bezoarticus Linnaeus, 1758) from the Pantanal Brazil.

Science.gov (United States)

Silveira, Júlia A G; Rabelo, Elida M L; Lacerda, Ana C R; Borges, Paulo A L; Tomás, Walfrido M; Pellegrin, Aiesca O; Tomich, Renata G P; Ribeiro, Múcio F B

2013-06-01

Hemoparasites were surveyed in 60 free-living pampas deer Ozotoceros bezoarticus from the central area of the Pantanal, known as Nhecolândia, State of Mato Grosso do Sul, Brazil, through the analysis of nested PCR assays and nucleotide sequencing. Blood samples were tested for Babesia/Theileria, Anaplasma spp., and Trypanosoma spp. using nPCR assays and sequencing of the 18S rRNA, msp4, ITS, and cathepsin L genes. The identity of each sequence was confirmed by comparison with sequences from GenBank using BLAST software. Forty-six (77%) pampas deer were positive for at least one hemoparasite, according to PCR assays. Co-infection occurred in 13 (22%) animals. Based on the sequencing results, 29 (48%) tested positive for A. marginale. Babesia/Theileria were detected in 23 (38%) samples, and according to the sequencing results 52% (12/23) of the samples were similar to T. cervi, 13% (3/23) were similar to Babesia bovis, and 9% (2/23) were similar to B. bigemina. No samples were amplified with the primers for T. vivax, while 11 (18%) were amplified with the ITS primers for T. evansi. The results showed pampas deer to be co-infected with several hemoparasites, including species that may cause serious disease in cattle. Pampas deer is an endangered species in Brazil, and the consequences of these infections to their health are poorly understood. Copyright © 2013 Elsevier GmbH. All rights reserved.

Uveitis due to anaplasmosis in a newborn Holstein calf Uveíte por anaplasmose em bezerro da raça Holandesa

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J.L. Laus

2006-12-01

Full Text Available Relata-se o caso de um bovino com, aproximadamente, seis horas de vida, apresentando histórico de febre, apatia, inapetência e desconforto ocular. No exame oftálmico rotineiro, encontraram-se valores do teste da lágrima de Schirmer aumentados, diminuição da pressão intra-ocular, hemorragia conjuntival, uveíte anterior, edema corneal e injeção ciliar em ambos os olhos. Coletou-se amostra de sangue para realização de esfregaço sangüíneo, hemograma e bioquímica sérica. No esfregaço visibilizaram-se corpúsculos de Anaplasma marginale em hemácias parasitadas. Quanto à bioquímica sérica, não foram observadas alterações. Os mesmos exames foram realizados na mãe do animal tendo como resultado a positividade para A. marginale. Para o controle da uveíte foi utilizada uma única aplicação de 0,5ml de betametasona a 5%, por via subconjuntival, em cada um dos olhos. Para o tratamento da anaplasmose, seguiu-se o protocolo a base de 10mg/kg de oxitetraciclina, por via intramuscular profunda, a cada 12 horas. Diante os achados oftálmicos e a presença de corpúsculos de A. marginale em hemácias parasitadas, contatou-se, ser um caso de uveíte crônica secundaria a doença sistêmica, factível com Anaplasmose.

MSP-tool : A VBA-based software tool for the analysis of multispecimen paleointensity data

NARCIS (Netherlands)

Monster, Marilyn W L; de Groot, Lennart V.; Dekkers, Mark J.

2015-01-01

The multispecimen protocol (MSP) is a method to estimate the Earth's magnetic field's past strength from volcanic rocks or archeological materials. By reducing the amount of heating steps and aligning the specimens parallel to the applied field, thermochemical alteration and multi-domain effects are

Anaplasma phagocytophilum, Bartonella spp., haemoplasma species and Hepatozoon spp. in ticks infesting cats: a large-scale survey.

Science.gov (United States)

Duplan, Florent; Davies, Saran; Filler, Serina; Abdullah, Swaid; Keyte, Sophie; Newbury, Hannah; Helps, Chris R; Wall, Richard; Tasker, Séverine

2018-03-20

Ticks derived from cats have rarely been evaluated for the presence of pathogens. The aim of this study was to determine the prevalence of Anaplasma phagocytophilum, Bartonella spp., haemoplasma species and Hepatozoon spp. in ticks collected from cats in the UK. Five hundred and forty DNA samples extracted from 540 ticks collected from cats presenting to veterinarians in UK practices were used. Samples underwent a conventional generic PCR assay for detection of Hepatozoon spp. and real-time quantitative PCR assays for detection of Anaplasma phagocytophilum and three feline haemoplasma species and a generic qPCR for detection of Bartonella spp. Feline 28S rDNA served as an endogenous internal PCR control and was assessed within the haemoplasma qPCR assays. Samples positive on the conventional and quantitative generic PCRs were submitted for DNA sequencing for species identification. Feline 28S rDNA was amplified from 475 of the 540 (88.0%) ticks. No evidence of PCR inhibition was found using an internal amplification control. Of 540 ticks, 19 (3.5%) contained DNA from one of the tick-borne pathogens evaluated. Pathogens detected were: A. phagocytophilum (n = 5; 0.9%), Bartonella spp. (n = 7; 1.3%) [including Bartonella henselae (n = 3; 0.6%) and Bartonella clarridgeiae (n = 1; 0.2%)], haemoplasma species (n = 5; 0.9%), "Candidatus Mycoplasma haemominutum" (n = 3; 0.6%), Mycoplasma haemofelis (n = 1; 0.2%), "Candidatus Mycoplasma turicensis" (n = 1; 0.2%), Hepatozoon spp. (n = 2; 0.4%), Hepatozoon felis (n = 1; 0.2%) and Hepatozoon silvestris (n = 1; 0.2%). These data provide important information on the prevalence of tick-borne pathogens in ticks infesting cats, with the identification of haemoplasma species, A. phagocytophilum, H. felis and Bartonella spp. (including B. henselae and B. clarridgeiae). This study also documents the first report of H. silvestris in ticks collected from domestic cats.

Infections and risk factors for livestock with species of Anaplasma, Babesia and Brucella under semi-nomadic rearing in Karamoja Region, Uganda.

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Lolli, Chiara; Marenzoni, Maria Luisa; Strona, Paolo; Lappo, Pier Giorgio; Etiang, Patrick; Diverio, Silvana

2016-03-01

A survey was conducted to estimate the prevalence of Anaplasma, Babesia and Brucella spp. infections in cattle, goats and sheep in the Karamoja Region of Uganda and to identify possible risk factors existing in this semi-nomadic and pastoral area. Low cost laboratory tests were used to diagnose infections (Rose Bengal test for Brucella spp. antibodies and direct microscopic examination for Anaplasma and Babesia spp.). Multivariable logistic regression models were applied to identify possible risk factors linked to gender, animal species, age (only for cattle) and districts. A total of 3935 cattle, 729 goats and 306 sheep of five districts of the Karamoja Region were tested. Seroprevalence for Brucella was 9.2 % (CI, 95 %: 8.4-10), for Anaplasma 19.5 % (CI 95 %: 18.4-20.6) and for Babesia 16 % (CI 95 %: 15-17.1). Significant differences in infections prevalence were observed against risk factors associated with districts and species. Cattle were the species with higher risk of the infections. Female gender was identified as at risk only for Brucella spp. infection. Cattle more than one year old had greater likelihood to be Brucella seropositive. Co-infections of Anaplasma and Babesia spp. were statistically associated, especially in goats and sheep. Further studies to identify risk factors related to host species and geographical districts are needed. The influence on the semi-nomadic agro-pastoral system in Karamoja of animal raids and animal mixing should be further investigated. Findings were important to sensitize Karamojong undertaking measures on infection control, especially on cattle, which are their main source of food.

Genetic Diversity of Tick-Borne Rickettsial Pathogens; Insights Gained from Distant Strains

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Sebastián Aguilar Pierlé

2014-01-01

Full Text Available The ability to capture genetic variation with unprecedented resolution improves our understanding of bacterial populations and their ability to cause disease. The goal of the pathogenomics era is to define genetic diversity that results in disease. Despite the economic losses caused by vector-borne bacteria in the Order Rickettsiales, little is known about the genetic variants responsible for observed phenotypes. The tick-transmitted rickettsial pathogen Anaplasma marginale infects cattle in tropical and subtropical regions worldwide, including Australia. Genomic analysis of North American A. marginale strains reveals a closed core genome defined by high levels of Single Nucleotide Polymorphisms (SNPs. Here we report the first genome sequences and comparative analysis for Australian strains that differ in virulence and transmissibility. A list of genetic differences that segregate with phenotype was evaluated for the ability to distinguish the attenuated strain from virulent field strains. Phylogenetic analyses of the Australian strains revealed a marked evolutionary distance from all previously sequenced strains. SNP analysis showed a strikingly reduced genetic diversity between these strains, with the smallest number of SNPs detected between any two A. marginale strains. The low diversity between these phenotypically distinct bacteria presents a unique opportunity to identify the genetic determinants of virulence and transmission.

Spatial distribution of Anaplasma phagocytophilum and Hepatozoon canis in red foxes (Vulpes vulpes) in Hungary.

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Tolnai, Z; Sréter-Lancz, Z; Sréter, T

2015-07-01

In recent years, Ehrlichia canis and Hepatozoon canis transmitted by Rhipicephalus sanguineus were reported from Hungary. The aim of the present study was to reveal the spatial distribution pattern of pathogens transmitted by R. sanguineus in a sentinel species, red fox (Vulpes vulpes) in Hungary and to analyse the relationship of these patterns with landscape and climate by geographical information systems. Fox carcasses, representing 0.5% of the total fox population were randomly selected out of all the foxes of Hungary. The spleen samples of the animals were tested by real-time PCR for Anaplasma platys, Babesia vogeli, E. canis and H. canis infection. Positive results were confirmed by conventional PCR followed by sequencing. The prevalence of H. canis infection was 22.2% (95% CI=18.4-26.4%), and this parasite was detected in all areas including the mountain regions of Hungary. These findings indicate that other tick species or other transmission routes (oral and transplacental) might be in the background of the countrywide distribution of H. canis. Anaplasma platys was not found; nevertheless, the prevalence of Anaplasma phagocytophilum infection transmitted by Ixodes ricinus was 12.5% (95% CI=9.7-16.1%) in foxes. B. vogeli and E. canis infection was not detected. There was no correlation between environmental parameter values in the home range of foxes and A. phagocytophilum or H. canis infection, which is in line with that observed in the case of tick species infesting foxes in Hungary. The results of this study indicate that R. sanguineus, if present, might be rare in Hungary. Our baseline study can be used for future evaluation of the effect of climate change on the spreading and emergence of R. sanguineus transmitted pathogens in Hungary. Copyright © 2015 Elsevier GmbH. All rights reserved.

La sexualidad en los jóvenes de sectores marginales. Un informe de avance

OpenAIRE

Aigneren Aburto, José Miguel

2004-01-01

La sexualidad en los jovenes de sectores marginales. El problema es la sexualidad de los adolescentes en un mundo de pobreza e inequidad. Este estudio tiene como objetivo general describir los crudos productos sexuales de la miseria y los mitos que envenenan a la juventud marginal. Es un intento de retratar a aquellos adolescentes que conocen el sexo en la escuela del submundo que esconden los cordones de extrema pobreza del Medellín de hoy. Los resultados citados corresponden a realidades co...

Chemical display of pyrimidine bases flipped out by modification-dependent restriction endonucleases of MspJI and PvuRts1I families.

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Evelina Zagorskaitė

Full Text Available The epigenetic DNA modifications 5-methylcytosine (5mC and 5-hydroxymethylcytosine (5hmC in eukaryotes are recognized either in the context of double-stranded DNA (e.g., by the methyl-CpG binding domain of MeCP2, or in the flipped-out state (e.g., by the SRA domain of UHRF1. The SRA-like domains and the base-flipping mechanism for 5(hmC recognition are also shared by the recently discovered prokaryotic modification-dependent endonucleases of the MspJI and PvuRts1I families. Since the mechanism of modified cytosine recognition by many potential eukaryotic and prokaryotic 5(hmC "readers" is still unknown, a fast solution based method for the detection of extrahelical 5(hmC would be very useful. In the present study we tested base-flipping by MspJI- and PvuRts1I-like restriction enzymes using several solution-based methods, including fluorescence measurements of the cytosine analog pyrrolocytosine and chemical modification of extrahelical pyrimidines with chloroacetaldehyde and KMnO4. We find that only KMnO4 proved an efficient probe for the positive display of flipped out pyrimidines, albeit the method required either non-physiological pH (4.3 or a substitution of the target cytosine with thymine. Our results imply that DNA recognition mechanism of 5(hmC binding proteins should be tested using a combination of all available methods, as the lack of a positive signal in some assays does not exclude the base flipping mechanism.

Ticks circulate Anaplasma, Ehrlichia, Babesia and Theileria parasites in North of Iran.

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Bekloo, Ahmad Jafar; Bakhshi, Hasan; Soufizadeh, Ayoub; Sedaghat, Mohammad Mehdi; Bekloo, Romina Jafar; Ramzgouyan, Maryam Roya; Chegeni, Asadollah Hosseini; Faghihi, Faezeh; Telmadarraiy, Zakkyeh

2017-12-15

Ticks serve as important vectors of some pathogens of medical importance all over the world and identification of their rate of infection plays an important role for further control of diseases. In the current study, we investigated on ticks collected from north of Iran where raising and caring livestock are the main task of the people in order to find evidences of infection of Babesia, Theileria, Anaplasma and Ehrlichia microbial agents. Totally, 609 hard tick species from two genera Hyalomma and Rhipicephalus including; Hy. scupense, Hy. dromedarii, Hy. rufipes, Hy. marginatum, Hy. asiaticum, Hy. anatolicum, R. bursa, R. sanguineus and R. turanicus were identified. Molecular analysis revealed the presence of Anaplasma, Ehrlichia, Babesia and Theileria microorganism agents in all collected tick species except Hy. asiaticum and R. turanicus. To the best of our knowledge, this is the first report on identification of B. occultans in Hyalomma anatolicum and B. ovis in Hyalomma sp in Iran. Copyright © 2017 Elsevier B.V. All rights reserved.

Fibronectin gene polymorphisms and clinical manifestations of mixed cryoglobulinemic syndrome: increased risk of lymphoma associated to MspI DD and HaeIII AA genotypes

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C. Fabro

2011-09-01

Full Text Available Objective: To analyse FN gene polymorphisms in type II mixed cryoglobulinemic syndrome (MCsn, an immune-complex mediated systemic vasculitis linked to hepatitis C virus (HCV infection and characterized by rheumatoid factor (RF positive B-cell proliferation at high risk for the progression into non Hodgkin’s lymphoma (NHL. Methods: Samples from eighty-one patients, with MCsn (type II serum cryoglobulins and clinical signs of vasculitis were studied. Sixthy-five (65/81, 80.3% patients were HCV-positive. Twenty-one (25.9% patients had developed a B-cell NHL during the course of MCsn. Seventy-two patients with HCV-negative and MC-unrelated NHL and 110 healthy blood donors (HBDs were taken as controls. HaeIIIb and MspI FN gene polymorphisms were analysed by PCR and specific restriction enzyme digestions, following reported procedures. Plasma FN levels were analysed by ELISA, whenever possible. Results: HaeIIIb and MspI allele and genotype frequencies did not differ between MCsn patients and HBDs. Of note, the DD-MspI (OR=5.56; CI=1.67-18.51, p=0.0046 and the AA-HaeIIIb (OR=5.54; CI=1.64-18.76, p=0.0066 homozygosis appeared significantly and independently associated with the development of B-cell NHL in MCsn patients, with the HaeIIIb A allele possibly conferring an increased risk of NHL in the general population (OR=1.72, CI=1.128- 2.635, p=0.0133. In contrast, the major vasculitic manifestations, such as peripheral neuropathy, skin ulcers and glomerulonephritis tended to be associated with the counterpart MspI C allele. No association between FN plasma levels and FN genotypes was found. Conclusion: Genotyping for MspI and HaeIIIb FN gene polymorphisms may be clinically relevant to define the predisposition to the major clinical manifestations in MCsn.

Interactions of rat repetitive sequence MspI8 with nuclear matrix proteins during spermatogenesis

International Nuclear Information System (INIS)

Rogolinski, J.; Widlak, P.; Rzeszowska-Wolny, J.

1996-01-01

Using the Southwestern blot analysis we have studied the interactions between rat repetitive sequence MspI8 and the nuclear matrix proteins of rats testis cells. Starting from 2 weeks the young to adult animal showed differences in type of testis nuclear matrix proteins recognizing the MspI8 sequence. The same sets of nuclear matrix proteins were detected in some enriched in spermatocytes and spermatids and obtained after fractionation of cells of adult animal by the velocity sedimentation technique. (author). 21 refs, 5 figs

Genotype analysis of partial growth hormone gene (GH891│MspI in Pesisir cattle and Simmental-Pesisir crossbred cattle

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T. Hartatik

2018-03-01

Full Text Available Growth hormone gene that controls metabolism and body size of animals and polymorphism of the gene is known related to performance of livestock. Identification of gene polymorphisms was important to get early information to determine genetic markers associated with economically desirable traits. Genetic mapping is one of important characterizations in the selection process. Pesisir cattle is one of local cattle in Indonesia that are reared in West Sumatera and have become adaptive to local environment. The present study aimed to identify the genotype of Growth Hormone gene (GH891│MspI of the Pesisir cattle and crossbred Simmental-Pesisir cattle (designated as SimPes and relationship between growth performance (body weight and body size. The present study was conducted to 30 blood samples consisted of 15 Pesisir cattle and 15 SimPes cattle. Body weight and body size were measured at 12-18 months of age. The Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP method was used in this research. The results showed that there exist three variant genotypes (MspI-/-, MspI+/-, MspI+/+ and that allele frequencies of MspI- and MspI+ were 53.3% and 46.7%, respectively. Non-significant effect has found between GH’s genotype and growth traits (Body measurement and body weight in both of breed cattle. In conclusion, three variant genotypes (GH891│MspI in Pesisir and SimPes (Simmental x Pesisir cattle population were found and no significant effect on growth trait performance.

Comparative genomics and transcriptomics of trait-gene association

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Pierlé Sebastián

2012-11-01

Full Text Available Abstract Background The Order Rickettsiales includes important tick-borne pathogens, from Rickettsia rickettsii, which causes Rocky Mountain spotted fever, to Anaplasma marginale, the most prevalent vector-borne pathogen of cattle. Although most pathogens in this Order are transmitted by arthropod vectors, little is known about the microbial determinants of transmission. A. marginale provides unique tools for studying the determinants of transmission, with multiple strain sequences available that display distinct and reproducible transmission phenotypes. The closed core A. marginale genome suggests that any phenotypic differences are due to single nucleotide polymorphisms (SNPs. We combined DNA/RNA comparative genomic approaches using strains with different tick transmission phenotypes and identified genes that segregate with transmissibility. Results Comparison of seven strains with different transmission phenotypes generated a list of SNPs affecting 18 genes and nine promoters. Transcriptional analysis found two candidate genes downstream from promoter SNPs that were differentially transcribed. To corroborate the comparative genomics approach we used three RNA-seq platforms to analyze the transcriptomes from two A. marginale strains with different transmission phenotypes. RNA-seq analysis confirmed the comparative genomics data and found 10 additional genes whose transcription between strains with distinct transmission efficiencies was significantly different. Six regions of the genome that contained no annotation were found to be transcriptionally active, and two of these newly identified transcripts were differentially transcribed. Conclusions This approach identified 30 genes and two novel transcripts potentially involved in tick transmission. We describe the transcriptome of an obligate intracellular bacterium in depth, while employing massive parallel sequencing to dissect an important trait in bacterial pathogenesis.

Immunogenicity of a virosomally-formulated Plasmodium falciparum GLURP-MSP3 chimeric protein-based malaria vaccine candidate in comparison to adjuvanted formulations

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Tamborrini Marco

2011-12-01

Full Text Available Abstract Background In clinical trials, immunopotentiating reconstituted influenza virosomes (IRIVs have shown great potential as a versatile antigen delivery platform for synthetic peptides derived from Plasmodium falciparum antigens. This study describes the immunogenicity of a virosomally-formulated recombinant fusion protein comprising domains of the two malaria vaccine candidate antigens MSP3 and GLURP. Methods The highly purified recombinant protein GMZ2 was coupled to phosphatidylethanolamine and the conjugates incorporated into the membrane of IRIVs. The immunogenicity of this adjuvant-free virosomal formulation was compared to GMZ2 formulated with the adjuvants Montanide ISA 720 and Alum in three mouse strains with different genetic backgrounds. Results Intramuscular injections of all three candidate vaccine formulations induced GMZ2-specific antibody responses in all mice tested. In general, the humoral immune response in outbred NMRI mice was stronger than that in inbred BALB/c and C57BL/6 mice. ELISA with the recombinant antigens demonstrated immunodominance of the GLURP component over the MSP3 component. However, compared to the Al(OH3-adjuvanted formulation the two other formulations elicited in NMRI mice a larger proportion of anti-MSP3 antibodies. Analyses of the induced GMZ2-specific IgG subclass profiles showed for all three formulations a predominance of the IgG1 isotype. Immune sera against all three formulations exhibited cross-reactivity with in vitro cultivated blood-stage parasites. Immunofluorescence and immunoblot competition experiments showed that both components of the hybrid protein induced IgG cross-reactive with the corresponding native proteins. Conclusion A virosomal formulation of the chimeric protein GMZ2 induced P. falciparum blood stage parasite cross-reactive IgG responses specific for both MSP3 and GLURP. GMZ2 thus represents a candidate component suitable for inclusion into a multi-valent virosomal

Targeting ticks for control of selected hemoparasitic diseases of cattle.

Science.gov (United States)

Kocan, K M

1995-03-01

Development in and transmission of hemoparasites by tick vectors are phenomena closely synchronized with the tick feeding cycle. In all known life cycles, initial infection of tick tissues occurs in midgut epithelial cells and transmission is effected as ticks feed after parasites have developed and multiplied in salivary glands. Many factors reviewed affect development and transmission of hemoparasites by ticks including age of ticks, artificial temperature, climate and/or season, tick stage or sex, hemoparasite variation, concurrent infection of ticks with other pathogens, host cell susceptibility, transovarial transmission, effect of hemoparasites on tick biology, and the effect of infecting parasitemia level in cattle on infection rates in ticks. Four hemoparasites of cattle, Anaplasma marginale, Cowdria ruminantium, Theileria parva, and Babesia spp., are all dependent on ticks for biological transmission. Babesia is transmitted transovarially whereas the other three are transmitted transstadially. Mechanical transfer of infective blood via fomites and mouthparts of biting arthropods is also a major means of transmission for Anaplasma marginale but not of the others. Potential control methods for hemoparasites that target parasites as they are developing in their respective tick hosts include tick control, vaccines (against ticks and parasites), and drugs (against ticks and parasites). Successful application of control strategies will be dependent upon thorough understanding of parasite developmental cycles, biology of the tick vectors and the immune response of cattle to ticks and to hemoparasites. The most effective control measures will be those that are targeted against both ticks and the hemoparasites they vector.

Coinfection of sheep with Anaplasma, Theileria and Babesia species in the Kurdistan Region, Iraq.

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Renneker, S; Abdo, J; Bakheit, M A; Kullmann, B; Beyer, D; Ahmed, J; Seitzer, U

2013-11-01

Infections of small ruminants with Anaplasma, Theileria and Babesia species are widely distributed in the old world and are of great economic impact. In Iraq, data on disease occurrence in sheep caused by above-mentioned infectious agents are scarce. This study provides information on various haemoparasitic agents infecting sheep in the Kurdistan Region, Iraq, using molecular diagnostic tools. Altogether, 195 samples originating from three governorates in the Kurdistan Region, namely Duhok, Erbil and Sulaimaniya, were analysed. The following pathogens were identified: Anaplasma ovis (62.6%), Theileria ovis (14.35%), T. lestoquardi (7.7%), T. uilenbergi (5.6%) and Babesia ovis (1.5%). T. uilenbergi is detected for the first time in Iraq. Coinfection of sheep with different pathogens could be observed in this study, and it was found that 45 of 195 (23%) of the samples contained more than one pathogen. Even triple-positive samples were identified in 3% of the investigated animals. In conclusion, we confirm the coinfection of sheep with various haemoparasitic pathogen species in the Kurdistan Region of Iraq. Further investigations are needed to reveal the epidemiology of the diseases, the respective tick vectors, and, in the case of coinfection, pathogens' interaction and possible cross-protection. © 2013 Blackwell Verlag GmbH.

A specific DNA probe which identifies Babesia bovis in whole blood.

Science.gov (United States)

Petchpoo, W; Tan-ariya, P; Boonsaeng, V; Brockelman, C R; Wilairat, P; Panyim, S

1992-05-01

A genomic library of Babesia bovis DNA from the Mexican strain M was constructed in plasmid pUN121 and cloned in Escherichia coli. Several recombinants which hybridized strongly to radioactively labeled B. bovis genomic DNA in an in situ screening were selected and further analyzed for those which specifically hybridized to B. bovis DNA. It was found that pMU-B1 had the highest sensitivity, detecting 25 pg of purified B. bovis DNA, and 300 parasites in 10 microliters of whole infected blood, or 0.00025% parasitemia. pMU-B1 contained a 6.0 kb B. bovis DNA insert which did not cross-hybridize to Babesia bigemina, Trypanosoma evansi, Plasmodium falciparum, Anaplasma marginale, Boophilus microplus and cow DNA. In the Southern blot analysis of genomic DNA, pMU-B1 could differentiate between two B. bovis geographic isolates, Mexican strain M and Thai isolate TS4. Thus, the pMU-B1 probe will be useful in the diagnosis of Babesia infection in cattle and ticks, and in the differentiation of B. bovis strains.

Associação entre os polimorfismos HaeIII e MspI do gene para o receptor alfa de estrogênio e densidade mamográfica em mulheres após a menopausa Association between HaeIII and MspI polymorphisms of estrogen receptor alpha gene and mammographic density in post-menopausal women

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Eduardo Henrique de Moura Ramos

2006-10-01

Full Text Available OBJETIVO: Avaliar a presença dos polimorfismos HaeIII e MspI do gene para o receptor de estrogênio alfa, bem como fatores clínicos e suas possíveis associações com a densidade mamográfica em mulheres após a menopausa. MÉTODOS: Foram avaliadas 115 mulheres após a menopausa, não usuárias de terapia hormonal e sem lesão mamária clínica ou mamograficamente identificada. A densidade mamográfica foi determinada por três observadores independentes, tomando-se como base a classificação dos padrões mamográficos do ACR-BIRADS®, 2003 (duas avaliações subjetivas e uma computadorizada - Adobe Photoshop® 7.0. Amostras de raspado bucal foram obtidas para extração de DNA e em seguida foi realizada uma PCR-RFLP (Polymerase Chain Reation - Restriction Fragment Length Polymorphism para análise de polimorfismos presentes no íntron 1 e éxon 1 do gene do REalfa (HaeIII e MspI. RESULTADOS: O polimorfismo HaeIII foi encontrado em 43 (37,4% das 115 mulheres, ao passo que o MspI estava presente em 96 (83,5% das mesmas. Houve alto grau de concordância entre os três observadores na determinação da densidade mamográfica. Trinta e quatro (29,6% mulheres tinham mamas densas, e 81 (70,4%, mamas lipossubstituídas. CONCLUSÃO: Não houve associação entre o polimorfismo HaeIII do gene para o receptor de estrogênio alfa e densidade mamográfica (Fisher = 0,712. Houve associação próxima à significância estatística entre o polimorfismo MspI e densidade (Fisher = 0,098. Idade, paridade e índice de massa corporal mostraram-se associados com densidade mamográfica.PURPOSE: To assess the presence of estrogen receptor gene polymorphisms HaeIII and MspI as well as clinical factors, and their possible associations with high mammographic density in post-menopausal women. METHODS: One hundred and fifteen post-menopausal women, not in use of hormonal therapy and without clinical or mammographic lesions were evaluated. Three independent observers

Analysis of RET promoter CpG island methylation using methylation-specific PCR (MSP), pyrosequencing, and methylation-sensitive high-resolution melting (MS-HRM): impact on stage II colon cancer patient outcome.

Science.gov (United States)

Draht, Muriel X G; Smits, Kim M; Jooste, Valérie; Tournier, Benjamin; Vervoort, Martijn; Ramaekers, Chantal; Chapusot, Caroline; Weijenberg, Matty P; van Engeland, Manon; Melotte, Veerle

2016-01-01

Already since the 1990s, promoter CpG island methylation markers have been considered promising diagnostic, prognostic, and predictive cancer biomarkers. However, so far, only a limited number of DNA methylation markers have been introduced into clinical practice. One reason why the vast majority of methylation markers do not translate into clinical applications is lack of independent validation of methylation markers, often caused by differences in methylation analysis techniques. We recently described RET promoter CpG island methylation as a potential prognostic marker in stage II colorectal cancer (CRC) patients of two independent series. In the current study, we analyzed the RET promoter CpG island methylation of 241 stage II colon cancer patients by direct methylation-specific PCR (MSP), nested-MSP, pyrosequencing, and methylation-sensitive high-resolution melting (MS-HRM). All primers were designed as close as possible to the same genomic region. In order to investigate the effect of different DNA methylation assays on patient outcome, we assessed the clinical sensitivity and specificity as well as the association of RET methylation with overall survival for three and five years of follow-up. Using direct-MSP and nested-MSP, 12.0 % (25/209) and 29.6 % (71/240) of the patients showed RET promoter CpG island methylation. Methylation frequencies detected by pyrosequencing were related to the threshold for positivity that defined RET methylation. Methylation frequencies obtained by pyrosequencing (threshold for positivity at 20 %) and MS-HRM were 13.3 % (32/240) and 13.8 % (33/239), respectively. The pyrosequencing threshold for positivity of 20 % showed the best correlation with MS-HRM and direct-MSP results. Nested-MSP detected RET promoter CpG island methylation in deceased patients with a higher sensitivity (33.1 %) compared to direct-MSP (10.7 %), pyrosequencing (14.4 %), and MS-HRM (15.4 %). While RET methylation frequencies detected by nested-MSP

Violencia basada en género en zonas urbanas y urbano-marginales de Lima y Callao, 2007 - 2010

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Pedro Mascaro-Sánchez

2012-04-01

Full Text Available Objetivos: Determinar y comparar las características de la violencia en pobladores de zonas urbanas y urbano-marginales. Diseño: Estudio epidemiológico descriptivo. Institución: Escuela de Obstetricia, Facultad de Medicina, Universidad Nacional Mayor de San Marcos, Lima, Perú. Participantes: Mujeres de Lima y Callao. Intervenciones: Entre los años 2007 y 2010, se aplicó una entrevista estructurada a 3 971 mujeres de zonas urbanas y urbano-marginales de Lima y Callao. Se incluyó mujeres residentes de la zona, excluyéndose aquellas con problemas psicológicos evidentes y problemas del habla. Los datos fueron analizados con estadísticas descriptivas y con el estadístico chi cuadrado, significativo a un nivel p<0,05. Principales medidas de resultados: Violencia física. Resultados: De las entrevistadas, 39% tenía entre 20 y 24 años de edad. El 47% identificó violencia física, siendo similar en zonas urbanas y urbano-marginales. Los empujones y bofetadas fueron la forma más frecuente de violencia en ambas áreas. Con respecto al estudio de la violencia psicológica, fueron más frecuentes la utilización de gritos y celos por posesividad, también en ambas áreas. El gasto de la pareja sin la consulta fue ligeramente mayor para el área urbano marginal (49% que para el área urbana (42%. El intento de tener relaciones sexuales contra su voluntad fue el tipo de violencia más frecuente (37%, más en el área urbana (41% que en el área urbano marginal (34%. Conclusiones: Se evidenció episodios de violencia física, psicológica, económica y sexual similares para el área urbana y el área urbano-marginal, siendo la violencia sexual más frecuente en el área urbana.

Heterogeneous genetic diversity pattern in Plasmodium vivax genes encoding merozoite surface proteins (MSP) -7E, -7F and -7L.

Science.gov (United States)

Garzón-Ospina, Diego; Forero-Rodríguez, Johanna; Patarroyo, Manuel A

2014-12-13

The msp-7 gene has become differentially expanded in the Plasmodium genus; Plasmodium vivax has the highest copy number of this gene, several of which encode antigenic proteins in merozoites. DNA sequences from thirty-six Colombian clinical isolates from P. vivax (pv) msp-7E, -7F and -7L genes were analysed for characterizing and studying the genetic diversity of these pvmsp-7 members which are expressed during the intra-erythrocyte stage; natural selection signals producing the variation pattern so observed were evaluated. The pvmsp-7E gene was highly polymorphic compared to pvmsp-7F and pvmsp-7L which were seen to have limited genetic diversity; pvmsp-7E polymorphism was seen to have been maintained by different types of positive selection. Even though these copies seemed to be species-specific duplications, a search in the Plasmodium cynomolgi genome (P. vivax sister taxon) showed that both species shared the whole msp-7 repertoire. This led to exploring the long-term effect of natural selection by comparing the orthologous sequences which led to finding signatures for lineage-specific positive selection. The results confirmed that the P. vivax msp-7 family has a heterogeneous genetic diversity pattern; some members are highly conserved whilst others are highly diverse. The results suggested that the 3'-end of these genes encode MSP-7 proteins' functional region whilst the central region of pvmsp-7E has evolved rapidly. The lineage-specific positive selection signals found suggested that mutations occurring in msp-7s genes during host switch may have succeeded in adapting the ancestral P. vivax parasite population to humans.

Anaplasma phagocytophilum Infection Subverts Carbohydrate Metabolic Pathways in the Tick Vector, Ixodes scapularis

Czech Academy of Sciences Publication Activity Database

Cabezas Cruz, Alejandro; Alberdi, P.; Valdés, James J.; Villar, M.; de la Fuente, J.

2017-01-01

Roč. 7, 7 February (2017), č. článku 23. ISSN 2235-2988 EU Projects: European Commission(XE) 278976 - ANTIGONE Institutional support: RVO:60077344 Keywords : proteomics * transcriptomics * glucose metabolism * Ixodes scapularis * Anaplasma phagocytophilum Subject RIV: EB - Genetics ; Molecular Biology OBOR OECD: Biochemistry and molecular biology Impact factor: 4.300, year: 2016

Prevalence Survey of Selected Bovine Pathogens in Water Buffaloes in the North Region of Brazil

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Jenevaldo Barbosa da Silva

2014-01-01

Full Text Available Although the largest buffalo herd in the occident is in the north region of Brazil, few studies have been conducted to assess the prevalence of selected parasitic diseases in buffalo herd. The present study was therefore conducted to investigate the epidemiological of Toxoplasma gondii, Neospora caninum, Anaplasma marginale, Babesia bigemina, and Babesia bovis in water buffaloes in the north region of Brazil. A total of 4796 buffalo blood samples were randomly collected from five provinces and simultaneously analyzed by the IFAT and ELISA. The serological prevalence of T. gondii and N. caninum was 41.3% and 55.5% in ELISA and 35.7% and 48.8% in IFAT, respectively. The overall prevalence of A. marginale, B. bovis, and B. bigemina was 63%, 25%, and 21% by ELISA and 50.0%, 22.5%, and 18.8% by IFAT, respectively. This study shows valuable information regarding the serological survey of selected bovine pathogens in water buffaloes in the north region of Brazil which will likely be very beneficial for the management and control programs of this disease.

Structural, stability, dynamic and binding properties of the ALS-causing T46I mutant of the hVAPB MSP domain as revealed by NMR and MD simulations.

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Shixiong Lua

Full Text Available T46I is the second mutation on the hVAPB MSP domain which was recently identified from non-Brazilian kindred to cause a familial amyotrophic lateral sclerosis (ALS. Here using CD, NMR and molecular dynamics (MD simulations, we characterized the structure, stability, dynamics and binding capacity of the T46I-MSP domain. The results reveal: 1 unlike P56S which we previously showed to completely eliminate the native MSP structure, T46I leads to no significant disruption of the native secondary and tertiary structures, as evidenced from its far-UV CD spectrum, as well as Cα and Cβ NMR chemical shifts. 2 Nevertheless, T46I does result in a reduced thermodynamic stability and loss of the cooperative urea-unfolding transition. As such, the T46I-MSP domain is more prone to aggregation than WT at high protein concentrations and temperatures in vitro, which may become more severe in the crowded cellular environments. 3 T46I only causes a 3-fold affinity reduction to the Nir2 peptide, but a significant elimination of its binding to EphA4. 4 EphA4 and Nir2 peptide appear to have overlapped binding interfaces on the MSP domain, which strongly implies that two signaling networks may have a functional interplay in vivo. 5 As explored by both H/D exchange and MD simulations, the MSP domain is very dynamic, with most loop residues and many residues on secondary structures highly fluctuated or/and exposed to bulk solvent. Although T46I does not alter overall dynamics, it does trigger increased dynamics of several local regions of the MSP domain which are implicated in binding to EphA4 and Nir2 peptide. Our study provides the structural and dynamic understanding of the T46I-causing ALS; and strongly highlights the possibility that the interplay of two signaling networks mediated by the FFAT-containing proteins and Eph receptors may play a key role in ALS pathogenesis.

Canine infection with Borrelia burgdorferi, Dirofilaria immitis, Anaplasma spp. and Ehrlichia spp. in Canada, 2013-2014.

Science.gov (United States)

Herrin, Brian H; Peregrine, Andrew S; Goring, Jonas; Beall, Melissa J; Little, Susan E

2017-05-19

Canine test results generated by veterinarians throughout Canada from 2013-2014 were evaluated to assess the geographical distribution of canine infection with Borrelia burgdorferi, Dirofilaria immitis, Ehrlichia spp., and Anaplasma spp. The percent positive test results of 115,636 SNAP® 4Dx® Plus tests from dogs tested were collated by province and municipality to determine the distribution of these vector-borne infections in Canada. A total of 2,844/115,636 (2.5%) dogs tested positive for antibody to B. burgdorferi. In contrast, positive test results for D. immitis antigen and antibodies to Ehrlichia spp. and Anaplasma spp. were low, with less than 0.5% of dogs testing positive for any one of these three agents nationwide. Provincial seroprevalence for antibodies to B. burgdorferi ranged from 0.5% (Saskatchewan)-15.7% (Nova Scotia); the areas of highest percent positive test results were in proximity to regions in the USA considered endemic for Lyme borreliosis, including Nova Scotia (15.7%) and Eastern Ontario (5.1%). These high endemic foci, which had significantly higher percent positive test results than the rest of the nation (P Canada. Using dogs as sentinels for these pathogens can aid in recognition of the public and veterinary health threat that each pose.

A Multi-wavelength Study of an Isolated MSP Bow Shock

Science.gov (United States)

Romani, Roger W.; Slane, Patrick; Green, Andrew

2017-08-01

PSR J2124-3358 is the only single MSP known to sport an Halpha bow shock. This shock, now also seen in the UV, encloses an unusual X-ray pulsar wind nebula (PWN) with a long off-axis trail. Combining the X-ray and UV images with AAT/KOALA integral field spectroscopy of the Halpha emission, we have an unusually complete picture of the pulsar's (101 km/s transverse) motion and the latitudinal distribution of its wind flux. These images reveal the 3-D orientation of a hard-spectrum PWN jet and a softer equatorial outflow. Within the context of a thin shock model, we can constrain the total energy output of the pulsar and the neutron star moment of inertia. The IFU spectra show extreme Balmer dominance, which also constrains the nature of the UV shock emission.

Anaplasmose bovina: parâmetros clínicos e de patologia clínica em bezerros infectados experimentalmente.: parâmetros clínicos e de patologia clínica em bezerros infectados experimentalmente.

OpenAIRE

Leonardo Costa Tavares Coelho

2007-01-01

Avaliou-se o comportamento dos parâmetros de patologia clínica em cinco bezerros HPB, com idade entre 90 e 120 dias, inoculados com Anaplasma marginale. O período médio de incubação foi de 13 dias, a duração do período patente, quando foram realizadas as análises, foi de 14 dias e o período convalescente durou 10 dias. Foram avaliadas as alterações no equilíbrio ácido-básico, perfil eletrolítico, parâmetros bioquímicos e hematológicos. Os animais apresentaram anemia, inicialmente normocítica ...

Detection and identification of Anaplasma phagocytophilum, Borrelia burgdorferi, and Rickettsia helvetica in Danish Ixodes ricinus ticks

DEFF Research Database (Denmark)

Skarphédinsson, Sigurdur; Lyholm, Birgitte Fjendbo; Ljungberg, Marianne

2007-01-01

% of adult ticks. The difference in prevalence between Anaplasma and Borrelia in adult ticks supports the idea that their maintenance cycles in nature may be different. Ticks were also infected with Rickettsia helvetica. Our study indicates that A. phagocytophilum prevalence in ticks in Denmark is as high...

Antibody levels against GLURP R2, MSP1 block 2 hybrid and AS202.11 and the risk of malaria in children living in hyperendemic (Burkina Faso) and hypo-endemic (Ghana) areas

DEFF Research Database (Denmark)

Adu, Bright; Cherif, Mariama K; Bosomprah, Samuel

2016-01-01

therefore need to be evaluated against different malaria endemicity backgrounds. METHODS: The associations between antibody responses to the chimeric merozoite surface protein 1 block 2 hybrid (MSP1 hybrid), glutamate-rich protein region 2 (GLURP R2) and the peptide AS202.11, and the risk of malaria were...

MSP-Tool: a VBA-based software tool for the analysis of multispecimen paleointensity data

Science.gov (United States)

Monster, Marilyn; de Groot, Lennart; Dekkers, Mark

2015-12-01

The multispecimen protocol (MSP) is a method to estimate the Earth's magnetic field's past strength from volcanic rocks or archeological materials. By reducing the amount of heating steps and aligning the specimens parallel to the applied field, thermochemical alteration and multi-domain effects are minimized. We present a new software tool, written for Microsoft Excel 2010 in Visual Basic for Applications (VBA), that evaluates paleointensity data acquired using this protocol. In addition to the three ratios (standard, fraction-corrected and domain-state-corrected) calculated following Dekkers and Böhnel (2006) and Fabian and Leonhardt (2010) and a number of other parameters proposed by Fabian and Leonhardt (2010), it also provides several reliability criteria. These include an alteration criterion, whether or not the linear regression intersects the y axis within the theoretically prescribed range, and two directional checks. Overprints and misalignment are detected by isolating the remaining natural remanent magnetization (NRM) and the partial thermoremanent magnetization (pTRM) gained and comparing their declinations and inclinations. The NRM remaining and pTRM gained are then used to calculate alignment-corrected multispecimen plots. Data are analyzed using bootstrap statistics. The program was tested on lava samples that were given a full TRM and that acquired their pTRMs at angles of 0, 15, 30 and 90° with respect to their NRMs. MSP-Tool adequately detected and largely corrected these artificial alignment errors.

Marine spatial planning (MSP: A first step to ecosystem-based management (EBM in the Wider Caribbean

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John C Ogden

2010-10-01

Full Text Available The rapid decline of coastal ecosystems of the Wider Caribbean is entering its fifth decade. Some of the best science documenting this decline and its causes has been done by the laboratories of the Association of Marine Laboratories of the Caribbean (AMLC. Alarmed at the trends, Caribbean conservation pioneers established marine protected areas (MPAs which spread throughout the region. Unfortunately, many have little or no protection and are now known to be too small to be effective in sustaining coastal ecosystems. Marine spatial planning (MSP holds much promise to encompass the large geographic scales of the ecological processes and human impacts that influence coastal ecosystems and adjacent lands. The AMLC, through the scientific expertise and the national political connections of its member institutions, is well-positioned to help implement a pilot project. MSP a first step in ecosystem-based management and has had considerable success elsewhere. It holds our best chance of sustaining human use and conserving the coral reefs and associated ecosystems. Rev. Biol. Trop. 58 (Suppl. 3: 71-79. Epub 2010 October 01.La rápida disminución de los ecosistemas costeros del Mar Caribe está entrando en su quinta década. Algunos de los mejores aportes científicos que documentan este descenso y sus causas han sido realizados por los laboratorios de la Asociación de Laboratorios Marinos del Caribe (ALMC. Alarmados por las tendencias, los pioneros de la conservación del Caribe establecieron áreas marinas protegidas (MPAs que se extendieron por toda la región. Desafortunadamente, muchas de estas áreas tienen poca o ninguna protección y ahora se conoce que son demasiado pequeñas para ser efectivas en el mantenimiento de los ecosistemas costeros. La planificación espacial marina (MSP es promisoria para englobar las grandes escalas geográficas de los procesos ecológicos y los impactos humanos que influyen en los ecosistemas costeros y las

Molecular detection of Anaplasma phagocytophilum DNA in the lesser horseshoe bat (Rhinolophus hipposideros) guano.

Science.gov (United States)

Afonso, E; Goydadin, A-C

2018-05-30

Although bats are increasingly recognised as potential reservoir hosts of human zoonotic pathogens, bacteria in bats are still poorly studied. To investigate the DNA faecal prevalence of the bacterium Anaplasma phagocytophilum, we sampled 23 lesser horseshoe bat (Rhinolophus hipposideros) maternity colonies located in buildings (churches, barns) in rural villages of eastern France. A total of 552 faecal samples were collected from 278 individuals. Anaplasma phagocytophilum DNA was detected in the faeces of 63 individuals (22.7%). Such high prevalence might suggest persistent infection in bats and/or a frequent consumption of insect preys carrying bacteria. Faecal DNA prevalence varied highly among colonies but was not related to the colony size. Faecal DNA prevalence was the highest in the Jura Department, where the density of ticks is known to be the highest across the study area. Because the sampled bats live in close proximity to humans, we discuss how concerning the presence of A. phagocytophilum DNA in bat guano is for humans frequenting places of worship that shelter bats. We also advocate future research to understand what a high faecal DNA prevalence in bat guano really implicates in terms of bacteria transmission.

Prevalence of Anaplasma phagocytophilum infection in feral cats in Massachusetts.

Science.gov (United States)

Galemore, Erin R; Labato, Mary A; O'Neil, Elizabeth

2018-01-01

The primary objective of this study was to determine the prevalence of Anaplasma phagocytophilum infection and exposure in adult feral cats in Massachusetts, an endemic area for A phagocytophilum and its tick vector Ixodes scapularis . The secondary objective was to determine if there were correlations between A phagocytophilum infection and the presence of anemia and thrombocytopenia. Blood samples were collected between June and December 2015 from 175 apparently healthy adult feral cats that were presented to trap and release spay/neuter centers in Massachusetts. Complete blood count, blood smear evaluation, SNAP 4Dx Plus test (IDEXX) and A phagocytophilum PCR were performed on all samples to document acute infection (PCR-positive and/or inclusions observed on blood smear) and exposure to A phagocytophilum (SNAP 4Dx Plus-positive for A phagocytophilum antibodies). The prevalence of exposure to A phagocytophilum in feral cats in Massachusetts was 9.7%, whereas the prevalence of acute infection was 6.9%. All blood smears were negative for Anaplasma species inclusions; therefore, acute infection was defined as testing positive on PCR analysis. No statistically significant correlations were identified for cats that were positive for A phagocytophilum on PCR analysis or SNAP 4Dx Plus test and the presence of anemia or thrombocytopenia. The prevalence of A phagocytophilum exposure in feral cats approaches 10% and is higher than the previously reported national average prevalence of 4.3% in the USA. A phagocytophilum infection may be an emerging infectious disease in cats. Further research is needed to determine the prevalence of clinical illness associated with A phagocytophilum infection in cats living in endemic areas.

OPERATING SYSTEM FOR WIRELESS SENSOR NETWORKS AND AN EXPERIMENT OF PORTING CONTIKIOS TO MSP430 MICROCONTROLLER

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Thang Vu Chien

2012-07-01

Full Text Available Wireless Sensor Networks (WSNs consist of a large number of sensor nodes, and are used for various applications such as building monitoring, environment control, wild-life habitat monitoring, forest fire detection, industry automation, military, security, and health-care. Each sensor node needs an operating system (OS that can control the hardware, provide hardware abstraction to application software, and fill in the gap between applications and the underlying hardware. In this paper, researchers present OS for WSNs and an experiment of porting contikiOS to MSP430 microcontroller which is very popular in many hardware platforms for WSNs. Researchers begin by presenting the major issues for the design of OS for WSNs. Then, researchers examine some popular operating systems for WSNs including TinyOS, ContikiOS, and LiteOS. Finally, researchers present an experiment of porting ContikiOS to MSP430 microcontroller. Wireless Sensor Networks (WSNs terdiri dari sejumlah besar sensor nodes, dan digunakan untuk berbagai aplikasi seperti pemantauan gedung, pengendalian lingkungan, pemantauan kehidupan habitat liar, deteksi kebakaran hutan, otomatisasi industri, militer, keamanan, dan kesehatan. Setiap sensor nodememerlukan sistem operasi (SO yang dapat mengontrol hardware, menyediakan abstraksi hardware untuk aplikasi perangkat lunak, dan mengisi kesenjangan antara aplikasi dan hardware. Dalam penelitian ini, peneliti menyajikan SO untuk WSNs dan percobaan dari port contikiOS untuk MSP430 mikrokontroler yang sangat populer di platformhardware untuk WSNs. Peneliti memulai dengan menghadirkan isu utama yaitu desain SO untuk WSNs. Lalu, penelitimemeriksa beberapa sistem operasi populer untuk WSNs, termasuk TinyOS, ContikiOS, dan LiteOS. Akhirnya penelitimenyajikan sebuah percobaan dari port ContikiOS untuk MSP430 mikrokontroler.

Design of Tropical Flowers Environmental Parameters Wireless Monitoring System Based on MSP430

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Huang Jian-Qing

2016-01-01

Full Text Available Considering the importance of real-time monitoring tropical flower environment parameters, the paper designs a wireless monitoring system based on MSP430F149 for tropical flower growing parameters. The proposed system uses sensor nodes to obtain data of temperature, humidity and light intensity, sink node to collect data from sensor nodes through wireless sensor network, and monitoring center to process data downloaded from the sink node through RS232 serial port. The node hardware platform is composed of a MSP430F149 processor, AM2306 and NHZD10AI sensors used to adopt temperature, humidity and light intensity data, and an nRF905 RF chip used to receive and send data. The node software, operated in IAR Embedded Workbench, adopts C Language to do node data collection and process, wireless transmission and serial port communication. The software of monitoring center develops in VB6.0, which can provide vivid and explicit real-time monitoring platform for flower farmers.

Test of the sheep ked Melophagus ovinus (L) as a vector of Anaplasma ovis Lestoquard.

Science.gov (United States)

Zaugg, J L; Coan, M E

1986-05-01

The sheep ked Melophagus ovinus was evaluated as a possible vector of Anaplasma ovis. In 4 tests, 45 to 293 keds were transferred from sheep with acute anaplasmosis, low level parasitemia, or carrier state of anaplasmosis to individual splenectomized ewes. Keds were confined in stockinette cages attached to the dorsal midline of the recipient hosts for 10 days. Throughout the 90-day observation periods after the keds were transferred, no clinical, serologic, or hematologic evidence of anaplasmosis was detected in any of the ked-recipient ewes. Results indicate that sheep keds probably are not mechanical or biological vectors of the ovine anaplasmosis organism.

Babesia canis vogeli, Ehrlichia canis, and Anaplasma platys infection in a dog.

Science.gov (United States)

Al Izzi, Salah; Martin, Donald S; Chan, Roxanne Y Y; Leutenegger, Christian M

2013-12-01

A 12-month-old male neutered mixed breed dog was presented with a history of diarrhea, lethargy, emaciation, polydypsia, and sniffling. Physical examination findings included pale mucous membranes, increased heart and respiratory rates, and normal rectal temperature (38°C). Hematologic abnormalities included anemia and thrombocytopenia. Biochemical abnormalities included hypoalbuminemia, hyperbilirubinemia, and elevated ALP and ALT activities. A SNAP 4Dx test result was positive for Ehrlichia canis. Babesia canis vogeli organisms were found in the peripheral blood films, while morulae of E canis were not seen. Real-time polymerase chain reaction testing confirmed the presence of both B c vogeli and E canis organisms, and also was positive for Anaplasma platys infection. The dog recovered following treatment with doxycycline and imidocarb dipropionate, with normal hematology and biochemical profiles. © 2013 American Society for Veterinary Clinical Pathology.

Disaggregating Tropical Disease Prevalence by Climatic and Vegetative Zones within Tropical West Africa.

Science.gov (United States)

Beckley, Carl S; Shaban, Salisu; Palmer, Guy H; Hudak, Andrew T; Noh, Susan M; Futse, James E

2016-01-01

Tropical infectious disease prevalence is dependent on many socio-cultural determinants. However, rainfall and temperature frequently underlie overall prevalence, particularly for vector-borne diseases. As a result these diseases have increased prevalence in tropical as compared to temperate regions. Specific to tropical Africa, the tendency to incorrectly infer that tropical diseases are uniformly prevalent has been partially overcome with solid epidemiologic data. This finer resolution data is important in multiple contexts, including understanding risk, predictive value in disease diagnosis, and population immunity. We hypothesized that within the context of a tropical climate, vector-borne pathogen prevalence would significantly differ according to zonal differences in rainfall, temperature, relative humidity and vegetation condition. We then determined if these environmental data were predictive of pathogen prevalence. First we determined the prevalence of three major pathogens of cattle, Anaplasma marginale, Babesia bigemina and Theileria spp, in the three vegetation zones where cattle are predominantly raised in Ghana: Guinea savannah, semi-deciduous forest, and coastal savannah. The prevalence of A. marginale was 63%, 26% for Theileria spp and 2% for B. bigemina. A. marginale and Theileria spp. were significantly more prevalent in the coastal savannah as compared to either the Guinea savanna or the semi-deciduous forest, supporting acceptance of the first hypothesis. To test the predictive power of environmental variables, the data over a three year period were considered in best subsets multiple linear regression models predicting prevalence of each pathogen. Corrected Akaike Information Criteria (AICc) were assigned to the alternative models to compare their utility. Competitive models for each response were averaged using AICc weights. Rainfall was most predictive of pathogen prevalence, and EVI also contributed to A. marginale and B. bigemina prevalence

Disaggregating Tropical Disease Prevalence by Climatic and Vegetative Zones within Tropical West Africa.

Directory of Open Access Journals (Sweden)

Carl S Beckley

Full Text Available Tropical infectious disease prevalence is dependent on many socio-cultural determinants. However, rainfall and temperature frequently underlie overall prevalence, particularly for vector-borne diseases. As a result these diseases have increased prevalence in tropical as compared to temperate regions. Specific to tropical Africa, the tendency to incorrectly infer that tropical diseases are uniformly prevalent has been partially overcome with solid epidemiologic data. This finer resolution data is important in multiple contexts, including understanding risk, predictive value in disease diagnosis, and population immunity. We hypothesized that within the context of a tropical climate, vector-borne pathogen prevalence would significantly differ according to zonal differences in rainfall, temperature, relative humidity and vegetation condition. We then determined if these environmental data were predictive of pathogen prevalence. First we determined the prevalence of three major pathogens of cattle, Anaplasma marginale, Babesia bigemina and Theileria spp, in the three vegetation zones where cattle are predominantly raised in Ghana: Guinea savannah, semi-deciduous forest, and coastal savannah. The prevalence of A. marginale was 63%, 26% for Theileria spp and 2% for B. bigemina. A. marginale and Theileria spp. were significantly more prevalent in the coastal savannah as compared to either the Guinea savanna or the semi-deciduous forest, supporting acceptance of the first hypothesis. To test the predictive power of environmental variables, the data over a three year period were considered in best subsets multiple linear regression models predicting prevalence of each pathogen. Corrected Akaike Information Criteria (AICc were assigned to the alternative models to compare their utility. Competitive models for each response were averaged using AICc weights. Rainfall was most predictive of pathogen prevalence, and EVI also contributed to A. marginale and B

The Prostaglandin E2-EP3 Receptor Axis Regulates Anaplasma phagocytophilum-Mediated NLRC4 Inflammasome Activation

Czech Academy of Sciences Publication Activity Database

Wang, X.; Shaw, D.K.; Hammond, H.L.; Sutterwala, F.S.; Rayamajhi, M.; Shirey, K.A.; Perkins, D.J.; Bonventre, J.V.; Velayutham, T.S.; Evans, S.M.; Rodino, K.G.; VieBrock, L.; Scanlon, K.M.; Carbonetti, N.H.; Carlyon, J.A.; Miao, E.A.; McBride, J.W.; Kotsyfakis, Michalis; Pedra, J. H. F.

2016-01-01

Roč. 12, č. 8 (2016), č. článku e1005803. E-ISSN 1553-7374 Institutional support: RVO:60077344 Keywords : Rickettsial agents * Anaplasma phagocytophilum * prostaglandin E2-EP3 Receptor Axis Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 7.003, year: 2015

Ixodes scapularis Tick Cells Control Anaplasma phagocytophilum Infection by Increasing the Synthesis of Phosphoenolpyruvate from Tyrosine

Czech Academy of Sciences Publication Activity Database

Cabezas Cruz, Alejandro; Espinosa, P. J.; Obregon, D. A.; Alberdi, P.; de la Fuente, J.

2017-01-01

Roč. 7, AUG 17 (2017), č. článku 375. ISSN 2235-2988 Institutional support: RVO:60077344 Keywords : proteomics * transcriptomics * phosphoenolpyruvate * glycerol-3-phosphate * Ixodes scapularis * Anaplasma phagocytophilum Subject RIV: EE - Microbiology, Virology OBOR OECD: Microbiology Impact factor: 4.300, year: 2016

Expression of recombinant myostatin propeptide pPIC9K-Msp plasmid in Pichia pastoris.

Science.gov (United States)

Du, W; Xia, J; Zhang, Y; Liu, M J; Li, H B; Yan, X M; Zhang, J S; Li, N; Zhou, Z Y; Xie, W Z

2015-12-28

Myostatin propeptide can inhibit the biological activity of myostatin protein and promote muscle growth. To express myostatin propeptide in vitro with a higher biological activity, we performed codon optimization on the sheep myostatin propeptide gene sequence, and mutated aspartic acid-76 to alanine based on the codon usage bias of Pichia pastoris and the enhanced biological activity of myostatin propeptide mutant. Modified myostatin propeptide gene was cloned into the pPIC9K plasmid to form the recombinant plasmid pPIC9K-Msp. Recombinant plasmid pPIC9K-Msp was transformed into Pichia pastoris GS115 by electrotransformation. Transformed cells were screened, and methanol was used to induce expression. SDS-PAGE and western blotting were used to verify the successful expression of myostatin propeptide with biological activity in Pichia pastoris, providing the basis for characterization of this protein.

Molecular detection of Anaplasma platys infection in free-roaming dogs and ticks from Kenya and Ivory Coast

Czech Academy of Sciences Publication Activity Database

Matei, I.A.; D'Amico, G.; Yao, P.K.; Ionică, A.M.; Kanyari, P. W. N.; Daskalaki, A.A.; Dumitrache, M.O.; Sándor, A.D.; Gherman, C.M.; Qablan, M.; Modrý, David; Mihalca, A. D.

2016-01-01

Roč. 9, MAR 16 (2016), č. článku 157. ISSN 1756-3305 Institutional support: RVO:60077344 Keywords : Africa * Anaplasma platys * carnivores * Kenya * Ivory Coast Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.080, year: 2016

Deficiencia de Vitamina A y Estado Nutricional Antropométrico en niños MarginalesUrbanos y Rurales en el Estado Zulia, Venezuela

OpenAIRE

Amaya-Castellanos, Daisy; Viloria-Castejón, Haydée; Ortega, Pablo; Gómez, Gisela; Urrieta, Jesús R; Lobo, Pablo; Estévez, Jesús

2002-01-01

El presente estudio transversal fue realizado para estimar la prevalencia de deficiencia de vitamina A (DVA) y desnutrición proteico-energética en una población infantil (24 a 85 meses) de tres barrios marginales urbanos de Maracaibo Estado Zulia, Venezuela (n=173) y de una zona rural (n=34), vecina a Maracaibo, por medio del análisis del retinol sérico y mediciones antropométricas, aplicando puntuación Z a los indicadores Talla//Edad (ZT//E); Peso//Edad (ZP//E) y Peso//Talla (ZP//T), compara...

Passive smoking, Cyp1A1 gene polymorphism and dysmenorrhea

Science.gov (United States)

Liu, Hong; Yang, Fan; Li, Zhiping; Chen, Changzhong; Fang, Zhian; Wang, Lihua; Hu, Yonghua; Chen, Dafang

2007-01-01

Objective This study investigated whether the association between passive smoking exposure and dysmenorrhea is modified by two susceptibility genes, CYP1A1MspI and CYP1A1HincII. Methods This report includes 1645 (1124 no dysmenorrhea, 521 dysmenorrhea) nonsmoking and nondrinking newly wed female workers at Anqing, China between June 1997 and June 2000. Multiple logistic regression models were used to estimate the associations of passive smoking exposure and genetic susceptibility with dysmenorrhea, adjusting for perceived stress. Results When stratified by women genotype, the adjusted OR of dysmenorrhea was 1.6 (95%CI=1.3-2.1) for passive smoking group with Ile/Ile462 genotype, and 1.5 (95%CI=1.1-2.1) with C/C6235 genotype, compared to non passive smoking group, respectively. The data further showed that there was a significant combined effect between passive smoking and the CYP1A1 Msp1 C/C6235 and HincII Ile/Ile462 genotype (OR=2.6, 95%CI=1.3-5.2). Conclusion CYP1A1 MspI and HincII genotypes modified the association between passive smoking and dysmenorrhea. PMID:17566695

VAPB/ALS8 MSP ligands regulate striated muscle energy metabolism critical for adult survival in caenorhabditis elegans.

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Sung Min Han

Full Text Available Mutations in VAPB/ALS8 are associated with amyotrophic lateral sclerosis (ALS and spinal muscular atrophy (SMA, two motor neuron diseases that often include alterations in energy metabolism. We have shown that C. elegans and Drosophila neurons secrete a cleavage product of VAPB, the N-terminal major sperm protein domain (vMSP. Secreted vMSPs signal through Roundabout and Lar-like receptors expressed on striated muscle. The muscle signaling pathway localizes mitochondria to myofilaments, alters their fission/fusion balance, and promotes energy production. Here, we show that neuronal loss of the C. elegans VAPB homolog triggers metabolic alterations that appear to compensate for muscle mitochondrial dysfunction. When vMSP levels drop, cytoskeletal or mitochondrial abnormalities in muscle induce elevated DAF-16, the Forkhead Box O (FoxO homolog, transcription factor activity. DAF-16 promotes muscle triacylglycerol accumulation, increases ATP levels in adults, and extends lifespan, despite reduced muscle mitochondria electron transport chain activity. Finally, Vapb knock-out mice exhibit abnormal muscular triacylglycerol levels and FoxO target gene transcriptional responses to fasting and refeeding. Our data indicate that impaired vMSP signaling to striated muscle alters FoxO activity, which affects energy metabolism. Abnormalities in energy metabolism of ALS patients may thus constitute a compensatory mechanism counterbalancing skeletal muscle mitochondrial dysfunction.

A novel merozoite surface antigen of Plasmodium falciparum (MSP-3 identified by cellular-antibody cooperative mechanism antigenicity and biological activity of antibodies

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Claude Oeuvray

1994-01-01

Full Text Available We report the identification of a 48kDa antigen targeted by antibodies which inhibit Plasmodium falciparum in vitro growth by cooperation with blood monocytes in an ADCI assay correlated to the naturally acquired protection. This protein is located on the surface of the merozoite stage of P. falciparum, and is detectable in all isolates tested. Epidemiological studies demonstrated that peptides derived from the amino acid sequence of MSP-3 contain potent B and T-cell epitopes recognized by a majority of individuals living in endemic areas. Moreover human antibodies either purified on the recombinant protein, or on the synthetic peptide MSP-3b, as well as antibodies raised in mice, were all found to promote parasite killing mediated by monocytes.

Molecular Detection of Anaplasma bovis in Cattle from Central Part of Iran

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Vahid Noaman

2010-09-01

Full Text Available Anaplasma bovis is a leukocytotropic agent of bovine anaplasmosis and there is no available information about molecular study on this agent in cattle of Iran. In this study a total 150 cattle blood samples were collected from central part of Iran. The presence of A. bovis examined using light microscopic detection and species-specific nested polymerase chain reaction (nested-PCR based on 16S rRNA gene. Of the 150 cattle, 4 (2.66 % was positive for A. bovis by nested-PCR. These data is the first A. bovis DNA presence in cattle from central part of Iran.

Detection of Borrelia lusitaniae, Rickettsia sp. IRS3, Rickettsia monacensis, and Anaplasma phagocytophilum in Ixodes ricinus collected in Madeira Island, Portugal.

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de Carvalho, Isabel Lopes; Milhano, Natacha; Santos, Ana Sofia; Almeida, Victor; Barros, Silvia C; De Sousa, Rita; Núncio, Maria Sofia

2008-08-01

A total of 300 Ixodes ricinus ticks were tested by polymerase chain reaction (PCR) for the presence of Borrelia spp., Rickettsia spp., and Anaplasma phagocytophilum. Sequence analysis demonstrated 8 (2.7%) ticks infected with B. lusitaniae, 60 (20%) with Rickettsia spp., and 1 (0.3%) with A. phagocytophilum. Seven (2.3%) ticks were coinfected with B. lusitaniae and Rickettsia spp., 2 (0.6%) with R. monacensis, and 5 (1.7%) with Rickettsia sp. IRS3. The results of this study suggest simultaneous transmission of multiple tick-borne agents on Madeira Island, Portugal.

Lungworm seroprevalence in free-ranging harbour seals and molecular characterisation of marine mammal MSP

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Sophia Arlena Ulrich

2016-04-01

Full Text Available Harbour seals (Phoca vitulina are frequently infected with the lungworms Otostrongylus circumlitus and Parafilaroides gymnurus. The infection is often accompanied by secondary bacterial infections and can cause severe bronchopneumonia and even death in affected animals. Hitherto, the detection of lungworm infections was based on post mortem investigations from animals collected within stranding networks and a valid detection method for live free-ranging harbour seals was not available. Recently, an ELISA was developed for detecting lungworm antibodies in harbour seal serum, using major sperm protein (MSP of the bovine lungworm, Dictyocaulus viviparus as recombinant diagnostic antigen. To determine lungworm seroprevalence in free-ranging harbour seals, serum was taken from four different seal age groups (n = 313 resulting in an overall prevalence of 17.9% (18.9% of males, 16.7% of females. 0.7% of harbour seals up to six weeks of age were seropositive, as were 89% of seals between six weeks and six months, 53.6% between six and 18 months and 24.2% of seals over 18 months of age. In the 18 months and over age group, seropositive animals showed statistically significant reductions in body weight (P = 0.003 and length (P < 0.001. Sera from lungworm infected harbour seals in rehabilitation (n = 6 revealed that duration of antibody persistence may be similar to that of lungworm infected cattle, but further studies are needed to confirm this. Phylogenetic analyses of MSP sequences of different marine and terrestrial mammal parasitic nematodes revealed that lungworm MSP of the genus Dictyocaulus (superfamily Trichostrongyloidea is more closely related to metastrongylid marine mammal lungworms than to trichostrongylid nematodes of terrestrial hosts.

Evaluación de la técnica de MSP-PCR para la caracterización molecular de aislamientos de Rhodotorula mucilaginosa provenientes de la Patagonia noroccidental Assessment of the MSP-PCR technique for the molecular characterization of Rhodotorula mucilaginosa isolates from northwestern Patagonia

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D. Libkind

2007-09-01

Full Text Available La identificación rápida de levaduras de origen ambiental o clínico es de importancia para el estudio de la biodiversidad de estos microorganismos y para la detección de posibles patógenos. Rhodotorula mucilaginosa es una levadura ubicua y pigmentada, capaz de producir infecciones en pacientes inmunocomprometidos. En este trabajo se evaluó la utilidad de la técnica de fingerprinting conocida como MSP-PCR (Micro/Minisatellite-Primed PCR en la caracterización e identificación de aislamientos ambientales de R. mucilaginosa provenientes de la Patagonia noroccidental. Sobre la base de sus caracteres fenotípicos, de un total de 200 levaduras pigmentadas se seleccionaron 110 aislamientos que presuntamente corresponderían a la especie R. mucilaginosa. Se evaluaron los iniciadores (GTG5, (GAC5 y M13 en aislamientos representativos, y se seleccionó el iniciador (GTG5 por ser el que permitió una mejor agrupación de los aislamientos pertenecientes a R. mucilaginosa y una mejor diferenciación de éstos con los de especies filogenéticamente próximas. Utilizando dicho iniciador, el 87% de los aislamientos de R. mucilaginosa presentó un perfil de MSP-PCR similar (> 60% al de la cepa de referencia CBS 316T de R. mucilaginosa. La técnica de MSP-PCR resultó efectiva, tanto para caracterizar e identificar un número elevado de aislamientos ambientales de R. mucilaginosa como para detectar polimorfismos en la especie.The rapid identification of environmental or clinical yeast isolates is important for biodiversity studies and the detection of probable pathogens. Rhodotorula mucilaginosa is a ubiquitous and pigmented yeast capable of infecting immunocompromised patients. In this study, we evaluated the Micro/mini satellite-primed PCR (MSP-PCR fingerprinting method for the characterization and identification of R. mucilaginosa isolates from natural environments in northwestern Patagonia. There were selected 110 putative R. mucilaginosa isolates

Plasmodium vivax Tryptophan Rich Antigen PvTRAg36.6 Interacts with PvETRAMP and PvTRAg56.6 Interacts with PvMSP7 during Erythrocytic Stages of the Parasite.

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Kriti Tyagi

Full Text Available Plasmodium vivax is most wide spread and a neglected malaria parasite. There is a lack of information on parasite biology of this species. Genome of this parasite encodes for the largest number of tryptophan-rich proteins belonging to 'Pv-fam-a' family and some of them are potential drug/vaccine targets but their functional role(s largely remains unexplored. Using bacterial and yeast two hybrid systems, we have identified the interacting partners for two of the P. vivax tryptophan-rich antigens called PvTRAg36.6 and PvTRAg56.2. The PvTRAg36.6 interacts with early transcribed membrane protein (ETRAMP of P.vivax. It is apically localized in merozoites but in early stages it is seen in parasite periphery suggesting its likely involvement in parasitophorous vacuole membrane (PVM development or maintenance. On the other hand, PvTRAg56.2 interacts with P.vivax merozoite surface protein7 (PvMSP7 and is localized on merozoite surface. Co-localization of PvTRAg56.2 with PvMSP1 and its molecular interaction with PvMSP7 probably suggest that, PvTRAg56.2 is part of MSP-complex, and might assist or stabilize the protein complex at the merozoite surface. In conclusion, the PvTRAg proteins have different sub cellular localizations and specific associated functions during intra-erythrocytic developmental cycle.

Quality assessment of DNA derived from up to 30 years old formalin fixed paraffin embedded (FFPE) tissue for PCR-based methylation analysis using SMART-MSP and MS-HRM.

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Kristensen, Lasse S; Wojdacz, Tomasz K; Thestrup, Britta B; Wiuf, Carsten; Hager, Henrik; Hansen, Lise Lotte

2009-12-21

The High Resolution Melting (HRM) technology has recently been introduced as a rapid and robust analysis tool for the detection of DNA methylation. The methylation status of multiple tumor suppressor genes may serve as biomarkers for early cancer diagnostics, for prediction of prognosis and for prediction of response to treatment. Therefore, it is important that methodologies for detection of DNA methylation continue to evolve. Sensitive Melting Analysis after Real Time - Methylation Specific PCR (SMART-MSP) and Methylation Sensitive - High Resolution Melting (MS-HRM) are two methods for single locus DNA methylation detection based on HRM. Here, we have assessed the quality of DNA extracted from up to 30 years old Formalin Fixed Paraffin Embedded (FFPE) tissue for DNA methylation analysis using SMART-MSP and MS-HRM. The quality assessment was performed on DNA extracted from 54 Non-Small Cell Lung Cancer (NSCLC) samples derived from FFPE tissue, collected over 30 years and grouped into five years intervals. For each sample, the methylation levels of the CDKN2A (p16) and RARB promoters were estimated using SMART-MSP and MS-HRM assays designed to assess the methylation status of the same CpG positions. This allowed for a direct comparison of the methylation levels estimated by the two methods for each sample. CDKN2A promoter methylation levels were successfully determined by SMART-MSP and MS-HRM in all 54 samples. Identical methylation estimates were obtained by the two methods in 46 of the samples. The methylation levels of the RARB promoter were successfully determined by SMART-MSP in all samples. When using MS-HRM to assess RARB methylation five samples failed to amplify and 15 samples showed a melting profile characteristic for heterogeneous methylation. Twenty-seven of the remaining 34 samples, for which the methylation level could be estimated, gave the same result as observed when using SMART-MSP. MS-HRM and SMART-MSP can be successfully used for single locus

Mixed Ehrlichia canis, Hepatozoon canis, and presumptive Anaplasma phagocytophilum infection in a dog.

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Mylonakis, Mathio E; Koutinas, Alex F; Baneth, Gad; Polizopoulou, Zoe; Fytianou, Anna

2004-01-01

A 5-month-old, female, mongrel dog was admitted to the Clinic of Companion Animal Medicine, Aristotle University of Thessaloniki, Greece, with depression, anorexia, fever, peripheral lymphadenopathy, splenomegaly, oculonasal discharge, nonregenerative anemia, and mild thrombocytopenia. Cytology of Giemsa-stained buffy coat, bone marrow, and lymph node aspiration smears revealed numerous morulae in mononuclear leukocytes and in neutrophils, and Hepatozoon canis gamonts in neutrophils. The dog was seropositive to Ehrlichia canis (immunofluorescence assay [IFA]) and Hepatozoon canis (ELISA) but not to Anaplasma phagocytophilum (IFA). A nested polymerase chain reaction performed on bone marrow aspirates was positive for E canis. This method was not applied for the detection of A phagocytophilum. Treatment with doxycycline and imidocarb dipropionate resulted in both clinical and parasitologic cure. This is the first reported case of a mixed infection with E canis, H canis, and presumptive A phagocytophilum. The findings emphasize the value of cytology in offering a quick and inexpensive diagnosis in mixed tick-borne infections of dogs.

Plasmodium vivax Tryptophan Rich Antigen PvTRAg36.6 Interacts with PvETRAMP and PvTRAg56.6 Interacts with PvMSP7 during Erythrocytic Stages of the Parasite

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Tyagi, Kriti; Hossain, Mohammad Enayet; Thakur, Vandana; Aggarwal, Praveen; Malhotra, Pawan; Mohmmed, Asif; Sharma, Yagya Dutta

2016-01-01

Plasmodium vivax is most wide spread and a neglected malaria parasite. There is a lack of information on parasite biology of this species. Genome of this parasite encodes for the largest number of tryptophan-rich proteins belonging to ‘Pv-fam-a’ family and some of them are potential drug/vaccine targets but their functional role(s) largely remains unexplored. Using bacterial and yeast two hybrid systems, we have identified the interacting partners for two of the P. vivax tryptophan-rich antigens called PvTRAg36.6 and PvTRAg56.2. The PvTRAg36.6 interacts with early transcribed membrane protein (ETRAMP) of P.vivax. It is apically localized in merozoites but in early stages it is seen in parasite periphery suggesting its likely involvement in parasitophorous vacuole membrane (PVM) development or maintenance. On the other hand, PvTRAg56.2 interacts with P.vivax merozoite surface protein7 (PvMSP7) and is localized on merozoite surface. Co-localization of PvTRAg56.2 with PvMSP1 and its molecular interaction with PvMSP7 probably suggest that, PvTRAg56.2 is part of MSP-complex, and might assist or stabilize the protein complex at the merozoite surface. In conclusion, the PvTRAg proteins have different sub cellular localizations and specific associated functions during intra-erythrocytic developmental cycle. PMID:26954579

Evaluation of an indirect ELISA for the diagnosis of Babesia bovis in Uruguay

International Nuclear Information System (INIS)

Cardozo, H.; Solari, M.A.; Etchebarne, J.

1992-01-01

In initially establishing the FAO/IAEA indirect ELISA for the detection of antibodies to Babesia bovis, the optical density (OD) values of sera from known positive or negative local cattle were compared to the OD values obtained from the negative and positive reference sera provided with the ELISA kit. The percentage of false positive and negative sera were 2.53% and 2.97% respectively. The cut-off values for the negative reference serum in the kit were compared with those of a local negative population. These values were found to be similar. The specificity of the test was evaluated by testing 30 sera from animals experimentally infected with Anaplasma marginale and 30 sera from animals infected with Babesia bigemina. These were no cross-reaction either between A. marginale and B. bovis or between B. bigemina and B. bovis. A serological survey using this ELISA kit was carried out on animals from an enzootic area and an area free from the vecot Boophilus microplus. 53 out of 282 animals (18.8%) in the enzootic area were positive whilst all the animals (113) from the free area were negative. This study would indicate that the FAO/IAEA ELISA kit has a sensitivity of around 98% and specificity of 97%. (author). 8 refs, 2 figs, 2 tabs

Prevalence of Dirofilaria immitis, Ehrlichia canis, Borrelia burgdorferi sensu lato, Anaplasma spp. and Leishmania infantum in apparently healthy and CVBD-suspect dogs in Portugal - a national serological study

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Cardoso Luís

2012-03-01

Full Text Available Abstract Background Canine vector-borne diseases (CVBDs are caused by a wide range of pathogens transmitted to dogs by arthropods including ticks and insects. Many CVBD-agents are of zoonotic concern, with dogs potentially serving as reservoirs and sentinels for human infections. The present study aimed at assessing the seroprevalence of infection with or exposure to Dirofilaria immitis, Ehrlichia canis, Borrelia burgdorferi sensu lato, Anaplasma spp. and Leishmania infantum in dogs in Portugal. Methods Based on 120 veterinary medical centres from all the regions of mainland and insular Portugal, 557 apparently healthy and 628 CVBD-suspect dogs were sampled. Serum, plasma or whole blood was tested for qualitative detection of D. immitis antigen and antibodies to E. canis, B. burgdorferi s. l., Anaplasma spp. and L. infantum with two commercial in-clinic enzyme-linked immunosorbent assay kits. Odds ratios (OR were calculated by logistic regression analysis to identify independent risk factors of exposure to the vector-borne agents. Results Total positivity levels to D. immitis, E. canis, B. burgdorferi, Anaplasma spp., L. infantum, one or more agents and mixed agents were 3.6%, 4.1%, 0.2%, 4.5%, 4.3%, 14.0% and 2.0% in the healthy group, and 8.9%, 16.4%, 0.5%, 9.2%, 25.2%, 46.3% and 11.6% in the clinically suspect group, respectively. Non-use of ectoparasiticides was a risk factor for positivity to one or more agents both in the apparently healthy (OR = 2.1 and CVBD-suspect (OR = 1.5 dogs. Seropositivity to L. infantum (OR = 7.6, E. canis (OR = 4.1 and D. immitis (OR = 2.4 were identified as risk factors for the presence of clinical signs compatible with CVBDs. Positivity to mixed agents was not found to be a risk factor for disease. Conclusions Dogs in Portugal are at risk of becoming infected with vector-borne pathogens, some of which are of zoonotic concern. CVBDs should be considered by practitioners and prophylactic measures must be put in

Safety and immunogenicity of a malaria vaccine, Plasmodium falciparum AMA-1/MSP-1 chimeric protein formulated in montanide ISA 720 in healthy adults.

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Jinhong Hu

Full Text Available BACKGROUND: The P. falciparum chimeric protein 2.9 (PfCP-2.9 consisting of the sequences of MSP1-19 and AMA-1 (III is a malaria vaccine candidate that was found to induce inhibitory antibodies in rabbits and monkeys. This was a phase I randomized, single-blind, placebo-controlled, dose-escalation study to evaluate the safety and immunogenicity of the PfCP-2.9 formulated with a novel adjuvant Montanide ISA720. Fifty-two subjects were randomly assigned to 4 dose groups of 10 participants, each receiving the test vaccine of 20, 50, 100, or 200 microg respectively, and 1 placebo group of 12 participants receiving the adjuvant only. METHODS AND FINDINGS: The vaccine formulation was shown to be safe and well-tolerated, and none of the participants withdrew. The total incidence of local adverse events (AEs was 75%, distributed among 58% of the placebo group and 80% of those vaccinated. Among the vaccinated, 65% had events that were mild and 15% experienced moderate AEs. Almost all systemic adverse reactions observed in this study were graded as mild and required no therapy. The participants receiving the test vaccine developed detectable antibody responses which were boosted by the repeated vaccinations. Sixty percent of the vaccinated participants had high ELISA titers (>1:10,000 of antigen-specific antibodies which could also recognize native parasite proteins in an immunofluorescence assay (IFA. CONCLUSION: This study is the first clinical trial for this candidate and builds on previous investigations supporting PfCP-2.9/ISA720 as a promising blood-stage malaria vaccine. Results demonstrate safety, tolerability (particularly at the lower doses tested and immunogenicity of the formulation. Further clinical development is ongoing to explore optimizing the dose and schedule of the formulation to decrease reactogenicity without compromising immunogenicity. TRIAL REGISTRATION: Chinese State Food and Drug Administration (SFDA 2002SL0046; Controlled

Estimates of repeatability and correlations of hemoparasites infection levels for cattle reared in endemic areas for Rhipicephalus microplus.

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Giglioti, Rodrigo; de Oliveira, Henrique Nunes; Bilhassi, Talita Barban; Portilho, Amanda Izeli; Okino, Cintia Hiromi; Marcondes, Cintia Righetti; de Sena Oliveira, Marcia Cristina

2018-01-30

Rhipicephalus microplus is a vector of cattle tick fever, a disease caused by the protozoans Babesia bovisand B. bigemina, and also anaplasmosis, produced by the Rickettsiales Anaplasma marginale. These tick-borne pathogens cause considerable losses to Brazilian livestock breeders and represent an obstacle to the expanded use of taurine breeds due to their higher sensitivity to ticks and hemoparasites compared to zebu breeds. Differences in the susceptibility to hemoparasites were also verified within breeds, suggesting that may be possible to select a most resistant phenotype. Therefore, repeatability of R. microplus counts and copy number of hemoparasites DNA were estimated, along with correlations between themselves, aiming to verify if those measures can be used as parameters to classify animals according to their parasite resistance degrees. Forty-two Canchim females kept on pastures naturally infested by ticks were evaluated for the level of infestation by R. microplus and infection by B. bovis, B. bigemina, and A. marginale. Twenty-four evaluations were performed once a month, for adult female ticks counts and blood samplings. The experimental period was divided into four phases, according to the animals age range: Phase 1: 8 to 13 months (collections 1 to 6); phase 2: 14 to 19 months (collections 7 to 12); phase 3: 20 to 25 months (collections 13 to 18), and phase 4: 26 to 31 months (collections 19 to 24). Blood samples were submitted to absolute quantification of hemoparasites DNA sequences using qPCR. The hemoparasite and tick counts data were transformed for normalization and were analyzed using mixed models. Among three species of hemoparasites studied, A. marginale presented the highest level of infection. During phase 3, B. bigemina presented higher infection levels (p hemoparasites and ticks. Moreover, the animal age may be an important factor related to resistance against these pathogens. The data obtained shed more light on the resistance to

Anaplasma phagocytophilum inhibits human neutrophil apoptosis via upregulation of bfl-1, maintenance of mitochondrial membrane potential and prevention of caspase 3 activation.

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Ge, Yan; Yoshiie, Kiyotaka; Kuribayashi, Futoshi; Lin, Mingqun; Rikihisa, Yasuko

2005-01-01

The inhibition of neutrophil apoptosis plays a central role in human granulocytic anaplasmosis. Intracellular signalling pathways through which the obligatory intracellular bacterium Anaplasma phagocytophilum inhibits the spontaneous apoptosis of human peripheral blood neutrophils were investigated. bfl-1 mRNA levels in uninfected neutrophils after 12 h in culture were reduced to approximately 5-25% of 0 h levels, but remained high in infected neutrophils. The eukaryotic RNA synthesis inhibitor, actinomycin D, prevented the maintenance of bfl-1 mRNA levels by A. phagocytophilum. Differences in mcl-1, bax, bcl-w, bad or bak mRNA levels in infected versus uninfected neutrophils were not remarkable. By using mitochondrial fluorescent dyes, Mitotracker Red and JC-1, it was found that most uninfected neutrophils lost mitochondrial membrane potential after 10-12 h incubation, whereas A. phagocytophilum-infected neutrophils maintained high membrane potential. Caspase 3 activity and the degree of apoptosis were lower in dose-dependent manner in A. phagocytophilum-infected neutrophils at 16 h post infection, as compared to uninfected neutrophils. Anti-active caspase 3 antibody labelling showed less positively stained population in infected neutrophils compared to those in uninfected neutrophils after 12 h incubation. These results suggest that A. phagocytophilum inhibits human neutrophil apoptosis via transcriptional upregulation of bfl-1 and inhibition of mitochondria-mediated activation of caspase 3.

Immunogenicity of a virosomally-formulated Plasmodium falciparum GLURP-MSP3 chimeric protein-based malaria vaccine candidate in comparison to adjuvanted formulations

DEFF Research Database (Denmark)

Tamborrini, Marco; Stoffel, Sabine A; Westerfeld, Nicole

2011-01-01

In clinical trials, immunopotentiating reconstituted influenza virosomes (IRIVs) have shown great potential as a versatile antigen delivery platform for synthetic peptides derived from Plasmodium falciparum antigens. This study describes the immunogenicity of a virosomally-formulated recombinant ...... fusion protein comprising domains of the two malaria vaccine candidate antigens MSP3 and GLURP....

A longitudinal study of type-specific antibody responses to Plasmodium falciparum merozoite surface protein-1 in an area of unstable malaria in Sudan

DEFF Research Database (Denmark)

Cavanagh, D R; Elhassan, I M; Roper, C

1998-01-01

Merozoite surface protein-1 (MSP-1) of Plasmodium falciparum is a malaria vaccine candidate Ag. Immunity to MSP-1 has been implicated in protection against infection in animal models. However, MSP-1 is a polymorphic protein and its immune recognition by humans following infection is not well unde...

An oven control system based on MCU MSP430F149

International Nuclear Information System (INIS)

Jing Lan; Zhu Haijun; Liu Caihong; Liu Yali; Gao Shiwei; Chinese Academia of Sciences, Beijing

2004-01-01

This paper introduces the structure, principle and feature of new generation of 16 bit and Flash-type microcontroller which belongs to the Texas Instruments MSP430F149 series with low power consumption and high performance. Details of the hardware structure and analysis software of the oven control system based on the microcontroller are expatiated. this systerm has the characteristics of strong function, simple struction, high reliability, strong noise resistances, no peripheral chip etc. The requirements of the temperature measurement and control system are fulfilled. (authors)

The software design of multi-branch, multi-point remote monitoring system for temperature measurement based on MSP430 and DS18B20

International Nuclear Information System (INIS)

Yu Jun; Yan Yu

2009-01-01

This paper present that the system can acquire the remote temperature measurement data of 40 monitoring points, through the RS-232 serial port and the Intranet. System's hardware is consist of TI's MSP430F149 mixed-signal processor and UA7000A network module. Using digital temperature sensor DS18B20, the structure is simple and easy to expand, the sensors directly send out the temperature data, MSP430F149 has the advantage of ultra-low-power and high degree of integration. Using msp430F149, the multi-branch multi-point temperature measurement system is powerful, simple structure, high reliability, strong anti-interference capability. The client software is user-friendly and easy to use, it is designed in Microsoft Visual C+ +6.0 environment. The monitoring system is able to complete a total of 4 branches of the 40-point temperature measurements in real-time remote monitoring. (authors)

DETECTION OF ANTIBODIES TO ANAPLASMA, BARTONELLA AND COXIELLA IN RURAL INHABITANTS OF THE CARIBBEAN AREA OF COLOMBIA

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Salim Máttar

2006-12-01

Full Text Available Objetivo. Establecer la seroprevalencia de Bartonella spp, Anaplasma phagocytophilum (antesErlichia y Coexiella burnetii. Materiales y métodos. Se analizaron sueros representativos de unsector de la población en el año 2003, recolectados de personas que trabajan en actividades delcampo en los departamentos de Córdoba y Sucre que sirvieron como población base de las muestrasque se obtuvieron. Los trabajadores rurales elegidos a participar tenían entra 16 – 65 años deedad. Los sueros fueron examinados por IFA para detección de anticuerpos contra IgG para Bartonellaspp, Erlichia Anaplasma phagocytophilum y Coexiella burnetii. Resultados. La seroprevalencia deanticuerpos de todos los microorganismos estudiados fue de 56.8%. De 81 muestras de sueroanalizadas el 26.6% fueron seropositivas contra C. burnetii, el 37.7% tuvieron anticuerpos contraBartonella y el 20% de los individuos evaluados fueron seropositivos para Anaplasmaphagocytophilum. Conclusiones. Nuestros datos indican que la prevalencia de anticuerpos contraBartonella, A. phagocytophilum y C. burnetii son altos en nuestra región. Los resultados indicanque estas enfermedades zoonoticas son muy comunes en las personas que residen en el área delcaribe colombiano. Este estudio demuestra por primera vez la presencia de estos microorganismosen Colombia.

REDES COMUNITARIAS: UN DESAFÍO PARA ESCUELAS URBANO-MARGINALES

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Mirta Giacobbe Tovani

2009-01-01

Full Text Available Las ideas básicas de este trabajo son conocer y comprender el entramado de la red de relaciones entre los distintos actores de las instituciones: escuela y organismos de la comunidad y su incidencia en la calidad educativa / calidad de vida. Analizamos la visión de los distintos actores educativos acerca de la inserción de la escuela en la comunidad. Tomamos como punto de partida el análisis de las relaciones entre calidad educativa y calidad de vida, en poblaciones urbano-marginales. Realizamos en primer lugar una investigación exploratoria, para conocer las características generales de escuelas que reciben población marginal. En una segunda etapa seleccionamos cuatro escuelas, que tomamos como casos e indagamos sus problemáticas. En ellas observamos gran dificultad para la construcción de redes comunitarias desde la institución escolar. Si bien responden a los problemas puntuales, mediante cambios o innovaciones planificados dinámica y flexiblemente, no existen soluciones duraderas debido a la falta de una convocatoria que comprometa a todas las instituciones de la comunidad constituyendo una efectiva relación en red. Estos resultados alcanzados en la investigación son producto de la triangulación de los datos estadísticos, documentales e históricos así como de los procedentes de la observación participante y de la información de las entrevistas en profundidad.

First evidence of Anaplasma platys and Hepatozoon canis co-infection in a dog from Romania--a case report.

Science.gov (United States)

Andersson, Martin; Turcitu, Mihai A; Stefanache, Mircea; Tamba, Paula; Barbuceanu, Florica; Chitimia, Lidia

2013-06-01

Anaplasma platys was first identified and described in North America as a Rickettsia-like, platelet-specific organism in dogs with infectious canine cyclic thrombocytopenia. In Europe, A. platys has so far mainly been described for some Mediterranean countries. Here, we describe a case of A. platys infection in a dog from Romania, confirmed by PCR. Additionally, the dog had a co-infection with Hepatozoon canis. To the best of our knowledge, this is the first case of A. platys infection in Romania and the first case of a co-infection with A. platys and H. canis altogether. Both pathogens should be considered as possible disease agents in dogs suffering from disease associated with tick bite in south-eastern Europe. Copyright © 2013 Elsevier GmbH. All rights reserved.

Prevalence of Anaplasma phagocytophilum infection and effect on lamb growth

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Steinshamn Håvard

2011-05-01

Full Text Available Abstract Background A major challenge in sheep farming during the grazing season along the coast of south-western Norway is tick-borne fever (TBF caused by the bacteria Anaplasma phagocytophilum that is transmitted by the tick Ixodes ricinus. Methods A study was carried out in 2007 and 2008 to examine the prevalence of A. phagocytophilum infection and effect on weaning weight in lambs. The study included 1208 lambs from farms in Sunndal Ram Circle in Møre and Romsdal County in Mid-Norway, where ticks are frequently observed. All lambs were blood sampled and serum was analyzed by an indirect fluorescent antibody assay (IFA to determine an antibody status (positive or negative to A. phagocytophilum infection. Weight and weight gain and possible effect of infection were analyzed using ANOVA and the MIXED procedure in SAS. Results The overall prevalence of infection with A. phagocytophilum was 55%. A lower weaning weight of 3% (1.34 kg, p A. phagocytophilum infection compared to seronegative lambs at an average age of 137 days. Conclusions The results show that A. phagocytophilum infection has an effect on lamb weight gain. The study also support previous findings that A. phagocytophilum infection is widespread in areas where ticks are prevalent, even in flocks treated prophylactic with acaricides.

Detection of Theileria annulata in blood samples of carrier cattle by PCR.

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d'Oliveira, C; van der Weide, M; Habela, M A; Jacquiet, P; Jongejan, F

1995-01-01

We report the detection of Theileria annulata, the causative agent of tropical theileriosis, by PCR in blood samples obtained from carrier cattle. The assay employs primers specific for the gene encoding the 30-kDa major merozoite surface antigen of T. annulata. A 721-bp fragment was amplified from blood samples taken monthly from calves experimentally infected with one of four different stocks of T. annulata originating in either Mauritania, Portugal, Spain, or Turkey. At the end of the experiment, five animals carried the infection for 12 months and two animals remained infected for 15 months. DNAs from six other Theileria species, T. parva, T. mutans, T. sergenti, T. buffeli, T. velifera, and T. taurotragi, were not amplified. Moreover, DNAs from four other hemoparasites (Anaplasma centrale, Anaplasma marginale, Babesia bovis, and Babesia bigemina) were also not amplified. As a control, primers derived from the small subunit rRNA gene of Theileria spp. amplified a 1.1-kb DNA fragment from all Theileria species examined but not from the other four hemoparasites. As few as two to three parasites per microliter of infected blood in a 50-microliters sample volume were detected by Southern or microplate hybridization with a T. annulata-specific cDNA probe. In addition, 92 field samples obtained from cattle in Spain were tested; 22% were positive in blood smears, 40% were positive by immunofluorescent antibody test, and 75% were positive for T. annulata by PCR. The method provides a useful diagnostic tool for detecting T. annulata carrier cattle. PMID:8567902

Molecular characterization and specific detection of Anaplasma species (AP-sd) in sika deer and its first detection in wild brown bears and rodents in Hokkaido, Japan.

Science.gov (United States)

Moustafa, Mohamed Abdallah Mohamed; Lee, Kyunglee; Taylor, Kyle; Nakao, Ryo; Sashika, Mariko; Shimozuru, Michito; Tsubota, Toshio

2015-12-01

A previously undescribed Anaplasma species (herein referred to as AP-sd) has been detected in sika deer, cattle and ticks in Japan. Despite being highly similar to some strains of A. phagocytophilum, AP-sd has never been detected in humans. Its ambiguous epidemiology and the lack of tools for its specific detection make it difficult to understand and interpret the prevalence of this Anaplasma species. We developed a method for specific detection, and examined AP-sd prevalence in Hokkaido wildlife. Our study included 250 sika deer (Cervus nippon yesoensis), 13 brown bears (Ursus arctos yesoensis) and 252 rodents including 138 (Apodemus speciosus), 45 (Apodemus argenteus), 42 (Myodes rufocanus) and 27 (Myodes rutilus) were collected from Hokkaido island, northern Japan, collected during 2010 to 2015. A 770 bp and 382 bp segment of the 16S rRNA and gltA genes, respectively, were amplified by nested PCR. Results were confirmed by cloning and sequencing of the positive PCR products. A reverse line blot hybridization (RLB) based on the 16S rRNA gene was then developed for the specific detection of AP-sd. The prevalence of AP-sd by nested PCR in sika deer was 51% (128/250). We detected this Anaplasma sp. for the first time in wild brown bears and rodents with a prevalence of 15% (2/13) and 2.4% (6/252), respectively. The sequencing results of the 16S rRNA and gltA gene amplicons were divergent from the selected A. phagocytophilum sequences in GenBank. Using a newly designed AP-sd specific probe for RLB has enabled us to specifically detect this Anaplasma species. Besides sika deer and cattle, wild brown bears and rodents were identified as potential reservoir hosts for AP-sd. This study provided a high throughput molecular method that specifically detects AP-sd, and which can be used to investigate its ecology and its potential as a threat to humans in Japan. Copyright © 2015 Elsevier B.V. All rights reserved.

Candidatus Neoehrlichia mikurensis and Anaplasma phagocytophilum: prevalences and investigations on a new transmission path in small mammals and ixodid ticks.

Science.gov (United States)

Obiegala, Anna; Pfeffer, Martin; Pfister, Kurt; Tiedemann, Tim; Thiel, Claudia; Balling, Anneliese; Karnath, Carolin; Woll, Dietlinde; Silaghi, Cornelia

2014-12-04

Small mammals are crucial for the life history of ixodid ticks, but their role and importance in the transmission cycle of tick-borne pathogens is mostly unknown. Candidatus Neoehrlichia mikurensis (CNM) and Anaplasma phagocytophilum are both tick-borne pathogens, and rodents are discussed to serve as main reservoir hosts for CNM but not for the latter especially in Germany. Analysing the prevalence of both pathogens in small mammals and their ticks in endemic regions may help to elucidate possible transmission paths in small mammal populations and between small mammals and ticks. In 2012 and 2013, small mammals were trapped at three different sites in Germany. DNA was extracted from different small mammal tissues, from rodent neonates, foetuses and from questing and attached ticks. DNA samples were tested for CNM and A. phagocytophilum by real-time PCR. Samples positive for A. phagocytophilum were further characterized at the 16S rRNA gene locus. CNM was detected in 28.6% of small mammals and in 2.2% of questing and 3.8% of attached ticks. Altogether 33 positive ticks were attached to 17 different hosts, while positive ticks per host ranged between one and seven. The prevalences for this pathogen differed significantly within small mammal populations comparing sites (χ(2): 13.3987; p: 0.0004) and between sexes. Male rodents had an approximately two times higher chance of infection than females (OR: 1.9652; 95% CI: 1.32-2.92). The prevalence for CNM was 31.8% (95% CI: 22-44) in rodent foetuses and neonates (23 of 67) from positive dams, and 60% (95% CI: 35.7-80.25) of positive gravid or recently parturient rodents (9 out of 15) had at least one positive foetus or neonate. Anaplasma phagocytophilum was detected at a low percentage in rodents (0-5.6%) and host-attached ticks (0.5-2.9%) with no significant differences between rodent species. However, attached nymphs were significantly more often infected than attached larvae (χ(2): 25.091; p: <0.0001). This study

Epigenetic silencing of host cell defense genes enhances intracellular survival of the rickettsial pathogen Anaplasma phagocytophilum.

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Jose C Garcia-Garcia

2009-06-01

Full Text Available Intracellular bacteria have evolved mechanisms that promote survival within hostile host environments, often resulting in functional dysregulation and disease. Using the Anaplasma phagocytophilum-infected granulocyte model, we establish a link between host chromatin modifications, defense gene transcription and intracellular bacterial infection. Infection of THP-1 cells with A. phagocytophilum led to silencing of host defense gene expression. Histone deacetylase 1 (HDAC1 expression, activity and binding to the defense gene promoters significantly increased during infection, which resulted in decreased histone H3 acetylation in infected cells. HDAC1 overexpression enhanced infection, whereas pharmacologic and siRNA HDAC1 inhibition significantly decreased bacterial load. HDAC2 does not seem to be involved, since HDAC2 silencing by siRNA had no effect on A. phagocytophilum intracellular propagation. These data indicate that HDAC up-regulation and epigenetic silencing of host cell defense genes is required for A. phagocytophilum infection. Bacterial epigenetic regulation of host cell gene transcription could be a general mechanism that enhances intracellular pathogen survival while altering cell function and promoting disease.

Anaplasma phagocytophilum in Danish sheep: confirmation by DNA sequencing

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Thamsborg Stig M

2009-12-01

Full Text Available Abstract Background The presence of Anaplasma phagocytophilum, an Ixodes ricinus transmitted bacterium, was investigated in two flocks of Danish grazing lambs. Direct PCR detection was performed on DNA extracted from blood and serum with subsequent confirmation by DNA sequencing. Methods 31 samples obtained from clinically normal lambs in 2000 from Fussingø, Jutland and 12 samples from ten lambs and two ewes from a clinical outbreak at Feddet, Zealand in 2006 were included in the study. Some of the animals from Feddet had shown clinical signs of polyarthritis and general unthriftiness prior to sampling. DNA extraction was optimized from blood and serum and detection achieved by a 16S rRNA targeted PCR with verification of the product by DNA sequencing. Results Five DNA extracts were found positive by PCR, including two samples from 2000 and three from 2006. For both series of samples the product was verified as A. phagocytophilum by DNA sequencing. Conclusions A. phagocytophilum was detected by molecular methods for the first time in Danish grazing lambs during the two seasons investigated (2000 and 2006.

AFCo1, a meningococcal B-derived cochleate adjuvant, strongly enhances antibody and T-cell immunity against Plasmodium falciparum merozoite surface protein 4 and 5.

Science.gov (United States)

Bracho, Gustavo; Zayas, Caridad; Wang, Lina; Coppel, Ross; Pérez, Oliver; Petrovsky, Nikolai

2009-02-27

Whilst a large number of malaria antigens are being tested as candidate malaria vaccines, a major barrier to the development of an effective vaccine is the lack of a suitable human adjuvant capable of inducing a strong and long lasting immune response. In this study, the ability of AFCo1, a potent T and B cell adjuvant based on cochleate structures derived from meningococcal B outer membrane proteoliposomes (MBOMP), to boost the immune response against two Plasmodium falciparum antigens, merozoite surface protein 4 (MSP4) and 5 (MSP5), was evaluated. Complete Freund's adjuvant (CFA), which is able to confer protection against malaria in animal MSP4/5 vaccine challenge models, was used as positive control adjuvant. MSP4 and 5-specific IgG, delayed-type hypersensitivity (DTH), T-cell proliferation, and cytokine production were evaluated in parallel in mice immunized three times intramuscularly with MSP4 or MSP5 incorporated into AFCo1, synthetic cochleate structures, CFA or phosphate buffered saline. AFCo1 significantly enhanced the IgG and T-cell response against MSP4 and MSP5, with a potency equivalent to CFA, with the response being characterized by both IgG1 and IgG2a isotypes, increased interferon gamma production and a strong DTH response, consistent with the ability of AFCo1 to induce Th1-like immune responses. Given the proven safety of MBOMP, which is already in use in a licensed human vaccine, AFCo1 could assist the development of human malaria vaccines that require a potent and safe adjuvant.

Low level of sequence diversity at merozoite surface protein-1 locus of Plasmodium ovale curtisi and P. ovale wallikeri from Thai isolates.

Science.gov (United States)

Putaporntip, Chaturong; Hughes, Austin L; Jongwutiwes, Somchai

2013-01-01

The merozoite surface protein-1 (MSP-1) is a candidate target for the development of blood stage vaccines against malaria. Polymorphism in MSP-1 can be useful as a genetic marker for strain differentiation in malarial parasites. Although sequence diversity in the MSP-1 locus has been extensively analyzed in field isolates of Plasmodium falciparum and P. vivax, the extent of variation in its homologues in P. ovale curtisi and P. ovale wallikeri, remains unknown. Analysis of the mitochondrial cytochrome b sequences of 10 P. ovale isolates from symptomatic malaria patients from diverse endemic areas of Thailand revealed co-existence of P. ovale curtisi (n = 5) and P. ovale wallikeri (n = 5). Direct sequencing of the PCR-amplified products encompassing the entire coding region of MSP-1 of P. ovale curtisi (PocMSP-1) and P. ovale wallikeri (PowMSP-1) has identified 3 imperfect repeated segments in the former and one in the latter. Most amino acid differences between these proteins were located in the interspecies variable domains of malarial MSP-1. Synonymous nucleotide diversity (πS) exceeded nonsynonymous nucleotide diversity (πN) for both PocMSP-1 and PowMSP-1, albeit at a non-significant level. However, when MSP-1 of both these species was considered together, πS was significantly greater than πN (pdiversity at this locus prior to speciation. Phylogenetic analysis based on conserved domains has placed PocMSP-1 and PowMSP-1 in a distinct bifurcating branch that probably diverged from each other around 4.5 million years ago. The MSP-1 sequences support that P. ovale curtisi and P. ovale wallikeri are distinct species. Both species are sympatric in Thailand. The low level of sequence diversity in PocMSP-1 and PowMSP-1 among Thai isolates could stem from persistent low prevalence of these species, limiting the chance of outcrossing at this locus.

Ehrlichia canis and Anaplasma platys in ticks of dogs in Cuiaba, Mato GrossoEhrlichia canis e Anaplasma platys em carrapatos de cães de Cuiabá, Mato Grosso

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Valéria Dutra

2012-06-01

Full Text Available Diseases transmitted by arthropods such as Rhipicephalus sanguineus, are caused by a spectrum of pathogens. Among these are the canine monocytic ehrlichiosis and cyclical thrombocytopenia with a cosmopolitan distribution. Aiming to verify the presence of DNA of Anaplasma platys and Ehrlichia canis in ticks R. sanguineus collected in the period 2008 to 2009 of 380 infected dogs. Ticks, after maceration, were subjected to DNA extraction and then nested PCR was performed for amplification of A. platys and E. canis. Of these, 81 (29.7% amplified DNA from ehrlichiais agents, where 38 (17.9% amplified in E. canis and 32 (15.7% for A. platys. The observation of two pathogens, combined with worldwide distribution of the tick R. sanguineus, demonstrates the high risk of infection with these pathogens in dogs in the city of Cuiaba. Doenças transmitidas por artrópodes, como o Rhipicephalus sanguineus, são causadas por um espectro de patógenos. Dentre estas, estão a erliquiose monocítica canina e trombocitopenia cíclica com distribuição cosmopolita. Com o objetivo de verificar a presença de DNA de Anaplasma platys e Ehrlichia canis em carrapatos R. sanguineus coletados no período de 2008 a 2009 de 380 cães infestados. Os carrapatos, após a maceração, foram submetidos a extração de DNA e, em seguida, foi realizada a Nested PCR para a amplificação da espécie A. platys e E. canis. Destes, 81 (29.7% amplificaram o DNA dos agentes ehrlichiais, onde 38 (17.9% amplificaram para E. canis e 32 (15.7% para A. platys. A observação dos dois patógenos, combinado com distribuição mundial do carrapato R. sanguineus, demonstra o elevado risco de infecção por esses patógenos de cães na cidade de Cuiabá.

Specific T-cell recognition of the merozoite proteins rhoptry-associated protein 1 and erythrocyte-binding antigen 1 of Plasmodium falciparum

DEFF Research Database (Denmark)

Jakobsen, P H; Hviid, L; Theander, T G

1993-01-01

The merozoite proteins merozoite surface protein 1 (MSP-1) and rhoptry-associated protein 1 (RAP-1) and synthetic peptides containing sequences of MSP-1, RAP-1, and erythrocyte-binding antigen 1, induced in vitro proliferative responses of lymphocytes collected from Ghanaian blood donors living i...... by individuals living in an area with a high transmission rate of malaria. Most of the donor plasma samples tested contained immunoglobulin G (IgG) and IgM antibodies recognizing the merozoite proteins, while only a minority showed high IgG reactivity to the synthetic peptides.......The merozoite proteins merozoite surface protein 1 (MSP-1) and rhoptry-associated protein 1 (RAP-1) and synthetic peptides containing sequences of MSP-1, RAP-1, and erythrocyte-binding antigen 1, induced in vitro proliferative responses of lymphocytes collected from Ghanaian blood donors living...

MODIFICACIÓN DEL ESQUELETO PEPTÍDICO DE FRAGMENTOS DE LA PROTEÍNA MSP-1 DEL Plasmodium falciparum: SÍNTESIS EN FASE SÓLIDA DE PSEUDOPÉPTIDOS AMIDA REDUCIDA

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José Manuel Lozano

2008-02-01

Full Text Available Los estadios asexuales sanguíneos del Plasmodium falciparum constituyen un blanco natural para el desarrollo de vacunas antimaláricas debido a la gran cantidad de antígenos expresados en su superficie que están implicados en el proceso de invasión del parásito a glóbulos rojos, desencadenando la enfermedaden el hospedero humano. Uno de estos antígenos es la proteína de superficie del merozoito MSP-1, una glicoproteína de 195 kDa en su forma madura.Recientemente se identificaron los péptidos de esta proteína con alta capacidad de unión a glóbulos rojos (GR. Entre todos ellos, el péptido sintético codificado 1585, cuya estructura primaria1282EVLYLKPLAGVYRSLKKQLE1301 pertenece al amino terminal del fragmento de 42 kDa de la proteína MSP-1, mostró propiedades de unión específica a GR. Posteriormente, se identificó el motivo de unión cuya secuencia de aminoácidos esLKP-A. Con el objeto de evaluar el papel del enlace peptídico en la modulación estructural del esqueleto peptídico se diseñaron pseudopéptidos amida reducida, mediante la sustitución de enlaces peptídicos normales de la forma [CO-NH] del motivo de unión a GR por el enlace Y[CH2NH]. De esta manera se obtuvieronformas monoméricas y poliméricas pseudopeptídicas del 1585. En este estudio reportamos algunas de las propiedades físicas y químicas de los pseudopéptidos análogos del 1585.

Anaplasma phagocytophilum in ticks in Slovenia

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Knap Nataša

2010-11-01

Full Text Available Abstract Ticks act as vectors of many pathogens of domestic animals and humans. Anaplasma phagocytophilum in Europe is transmitted by the ixodid tick vector Ixodes ricinus. A. phagocytophilum causes a disease with diverse clinical signs in various hosts. A great genetic diversity of the groESL operon of A. phagocytophilum has been found in ticks elsewhere. In Slovenia, the variety of the groESL operon was conducted only on deer samples. In this study, the prevalence of infected ticks was estimated and the diversity of A. phagocytophilum was evaluated. On 8 locations in Slovenia, 1924 and 5049 (6973 I. ricinus ticks were collected from vegetation in the years 2005 and 2006, respectively. All three feeding stages of the tick's life cycle were examined. The prevalence of ticks infected with A. phagocytophilum in the year 2005 and in the year 2006 was 0.31% and 0.63%, respectively, and it did not differ considerably between locations. The similarity among the sequences of groESL ranged from 95.6% to 99.8%. They clustered in two genetic lineages along with A. phagocytophilum from Slovenian deer. One sequence formed a separate cluster. According to our study, the prevalence of A. phagocytophilum in ticks is comparable to the findings in other studies in Europe, and it does not vary considerably between locations and tick stages. According to groESL operon analysis, two genetic lineages have been confirmed and one proposed. Further studies on other genes would be useful to obtain more information on genetic diversity of A. phagocytophilum in ticks in Slovenia.

Influence des paramètres d'un laser ER: YAG sur l'adaptation marginale d'une restauration à faible contraction

OpenAIRE

Peeters, Frédéric

2013-01-01

Quarante molaires ont été réparties en cinq groupes. C'est avec un laser de type Er:YAG que des cavités occlusales ont été faites sur les dents des quatre premiers groupes et cela avec certains paramètres définis et spécifiques à chacun. Les dents du dernier groupe quant à elles, ont subi un traitement conventionnel de fraisage. L’obturation des cavités a été faite avec le même matériau, une résine composite à faible taux de contraction. L'adaptation marginale de chaque dent a ensuite été com...

AFCo1, a meningococcal B-derived cochleate adjuvant, strongly enhances antibody and T-cell immunity against Plasmodium falciparum merozoite surface protein 4 and 5

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Pérez Oliver

2009-02-01

Full Text Available Abstract Background Whilst a large number of malaria antigens are being tested as candidate malaria vaccines, a major barrier to the development of an effective vaccine is the lack of a suitable human adjuvant capable of inducing a strong and long lasting immune response. In this study, the ability of AFCo1, a potent T and B cell adjuvant based on cochleate structures derived from meningococcal B outer membrane proteoliposomes (MBOMP, to boost the immune response against two Plasmodium falciparum antigens, merozoite surface protein 4 (MSP4 and 5 (MSP5, was evaluated. Methods Complete Freund's adjuvant (CFA, which is able to confer protection against malaria in animal MSP4/5 vaccine challenge models, was used as positive control adjuvant. MSP4 and 5-specific IgG, delayed-type hypersensitivity (DTH, T-cell proliferation, and cytokine production were evaluated in parallel in mice immunized three times intramuscularly with MSP4 or MSP5 incorporated into AFCo1, synthetic cochleate structures, CFA or phosphate buffered saline. Results AFCo1 significantly enhanced the IgG and T-cell response against MSP4 and MSP5, with a potency equivalent to CFA, with the response being characterized by both IgG1 and IgG2a isotypes, increased interferon gamma production and a strong DTH response, consistent with the ability of AFCo1 to induce Th1-like immune responses. Conclusion Given the proven safety of MBOMP, which is already in use in a licensed human vaccine, AFCo1 could assist the development of human malaria vaccines that require a potent and safe adjuvant.

DESENVOLVIMENTO DE ROBÔ DE SUMÔ MICROCONTROLADO POR MSP430

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Magno Melo

2017-02-01

Full Text Available Este artigo apresenta a pesquisa e definição dos componentes necessários ao desenvolvimento de um robô autônomo a ser utilizado em competição de sumô de robôs. Este tipo de equipamento, também conhecido como robô segue faixa, será desenvolvido com o objetivo de introdução à robótica, bem como do estudo do funcionamento e aplicabilidade do microcontrolador MSP430, o qual será o componente responsável pela automação deste robô. Assim, a partir desta definição, faz-se necessária a especificação dos demais componentes integrantes do equipamento (chassi, meio de locomoção, motores, sensores e alimentação de tal forma que o robô possa desempenhar adequadamente as tarefas para os quais o mesmo foi idealizado.

Methylation-associated down-regulation of RASSF1A and up-regulation of RASSF1C in pancreatic endocrine tumors

International Nuclear Information System (INIS)

Malpeli, Giorgio; Amato, Eliana; Dandrea, Mario; Fumagalli, Caterina; Debattisti, Valentina; Boninsegna, Letizia; Pelosi, Giuseppe; Falconi, Massimo; Scarpa, Aldo

2011-01-01

RASSF1A gene silencing by DNA methylation has been suggested as a major event in pancreatic endocrine tumor (PET) but RASSF1A expression has never been studied. The RASSF1 locus contains two CpG islands (A and C) and generates seven transcripts (RASSF1A-RASSF1G) by differential promoter usage and alternative splicing. We studied 20 primary PETs, their matched normal pancreas and three PET cell lines for the (i) methylation status of the RASSF1 CpG islands using methylation-specific PCR and pyrosequencing and (ii) expression of RASSF1 isoforms by quantitative RT-PCR in 13 cases. CpG island A methylation was evaluated by methylation-specific PCR (MSP) and by quantitative methylation-specific PCR (qMSP); pyrosequencing was applied to quantify the methylation of 51 CpGs also encompassing those explored by MSP and qMSP approaches. MSP detected methylation in 16/20 (80%) PETs and 13/20 (65%) normal pancreas. At qMSP, 11/20 PETs (55%) and 9/20 (45%) normals were methylated in at least 20% of RASSF1A alleles. Pyrosequencing showed variable distribution and levels of methylation within and among samples, with PETs having average methylation higher than normals in 15/20 (75%) cases (P = 0.01). The evaluation of mRNA expression of RASSF1 variants showed that: i) RASSF1A was always expressed in PET and normal tissues, but it was, on average, expressed 6.8 times less in PET (P = 0.003); ii) RASSF1A methylation inversely correlated with its expression; iii) RASSF1 isoforms were rarely found, except for RASSF1B that was always expressed and RASSF1C whose expression was 11.4 times higher in PET than in normal tissue (P = 0.001). A correlation between RASSF1A expression and gene methylation was found in two of the three PET cell lines, which also showed a significant increase in RASSF1A expression upon demethylating treatment. RASSF1A gene methylation in PET is higher than normal pancreas in no more than 75% of cases and as such it cannot be considered a marker for this neoplasm

Prevalencia de caries de infancia temprana en niños menores de 6 años de edad, residentes en poblados urbano marginales de Lima Norte

OpenAIRE

Villena Sarmiento, Rita; Pachas Barrionuevo, Flor; Sánchez Huamán, Yhedina; Carrasco Loyola, Milagros

2014-01-01

La prevalencia de caries de infancia temprana es un problema de salud pública prevenible y que afecta a un gran número de niños. El propósito de este trabajo fue determinar la prevalencia yseveridad de la caries dental en niños de 6-71 meses de edad de comunidades urbano marginales de Lima. Se evaluaron a 332 niños con los criterios de caries dental de la OMS, con equipo noinvasivo, bajo luz natural, y con técnica de rodilla-rodilla para los más pequeños. Fueron calibrados 3 odontólogos en el...

Genetic Diversity of Plasmodium falciparum Field Isolates in Central Sudan Inferred by PCR Genotyping of Merozoite Surface Protein 1 and 2.

Science.gov (United States)

Hamid, Muzamil M Abdel; Mohammed, Sara B; El Hassan, Ibrahim M

2013-02-01

Characterization of Plasmodium falciparum diversity is commonly achieved by amplification of the polymorphic regions of the merozoite surface proteins 1 (MSP1) and 2 (MSP2) genes. The present study aimed to determine the allelic variants distribution of MSP1 and MSP2 and multiplicity of infection in P. falciparum field isolates from Kosti, central Sudan, an area characterized by seasonal malaria transmission. Total 121 samples (N = 121) were collected during a cross-sectional survey between March and April 2003. DNA was extracted and MSP1 and MSP2 polymorphic loci were genotyped. The total number of alleles identified in MSP1 block 2 was 11, while 16 alleles were observed in MSP2 block 3. In MSP1, RO33 was found to be the predominant allelic type, carried alone or in combination with MAD20 and K1 types, whereas FC27 family was the most prevalent in MSP2. Sixty two percent of isolates had multiple genotypes and the overall mean multiplicity of infection was 1.93 (CI 95% 1.66-2.20). Age correlated with parasite density (P = 0.017). In addition, a positive correlation was observed between parasite densities and the number of alleles (P = 0.022). Genetic diversity in P. falciparum field isolates in central Sudan was high and consisted of multiple clones.

A putative marker for human pathogenic strains of Anaplasma phagocytophilum correlates with geography and host, but not human tropism.

Science.gov (United States)

Foley, Janet; Stephenson, Nicole; Cubilla, Michelle Pires; Qurollo, Barbara; Breitschwerdt, Edward B

2016-03-01

Anaplasma phagocytophilum is an Ixodes species tick-transmitted bacterium that is capable of infecting a variety of host species, although there is a diversity of bacterial strains with differing host tropism. Recent analysis of A. phagocytophilum strains suggested that "drhm", a gene locus designated "distantly related to human marker" (drhm), which was predicted to be an integral membrane protein with possible transporter functions was not present in available canine and human isolates. By assessing 117 strains from 14 host species from across the US, we extended this analysis. Phylogenetic clades were associated with geography, but not host species. Additionally, a virulent clade that lacks drhm and infects dogs, horses, and humans in northeastern US was identified. Copyright © 2015 Elsevier GmbH. All rights reserved.

A high frequency MspI RFLP at the human vitamin D binding protein (hDBP) locus

Energy Technology Data Exchange (ETDEWEB)

Ray, K; Cooke, N E [Univ. of Pennsylvania, Philadelphia (USA)

1988-07-25

A nearly full-length cDNA encoding the human vitamin D binding protein (hDBP), also known as Gc-globulin, was isolated from a human liver cDNA expression library. This 1.73 kb cDNA was digested with EcoRI and the 5{prime}, 140 bp fragment of the cDNA, subcloned into plasmid SP65 (phDBP140), was used as probe. MspI identifies a two allele polymorphism with either a band at 12 kb or a band at 5 kb. The frequency was estimated from a study of 24 unrelated North American Caucasians. The hDBP gene has been localized to chromosome 4 using a cDNA probe and a panel of rodent X human somatic cell hybrids. It was sublocalized to 4q11-q13 by in situ hybridization. Co-dominant autosomal segregation of the polymorphic alleles has been observed in two informative families (20 individuals). With overexposure of the autoradiograph two faint variant bands are seen at 17 kb and 13.5 kb which appear to cosegregate with the 12 kb band and the 5 kb band respectively.

Safety and immunogenicity of the malaria vaccine candidate MSP3 long synthetic peptide in 12-24 months-old Burkinabe children.

NARCIS (Netherlands)

Sirima, S.B.; Tiono, A.B.; Diarra, A.; Ouedraogo, A.L.; Yaro, J.B.; Ouedraogo, E.; Gansane, A.; Bougouma, E.C.; Konate, A.T.; Kabore, Y.; Traore, A.; Roma, C.; Soulama, I.; Luty, A.J.F.; Cousens, S.; Nebie, I.

2009-01-01

BACKGROUND: A Phase Ia trial in European volunteers of the candidate vaccine merozoite surface protein 3 (MSP3), a Plasmodium falciparum blood stage membrane, showed that it induces biologically active antibodies able to achieve parasite killing in vitro, while a phase Ib trial in semi-immune adult

Introduction to embedded systems using microcontrollers and the MSP430

CERN Document Server

Jiménez, Manuel; Couvertier, Isidoro

2014-01-01

This textbook serves as an introduction to the subject of embedded systems design, using microcontrollers as core components.  It develops concepts from the ground up, covering the development of embedded systems technology, architectural and organizational aspects of controllers and systems, processor models, and peripheral devices. Since microprocessor-based embedded systems tightly blend hardware and software components in a single application, the book also introduces the subjects of data representation formats, data operations, and programming styles. The practical component of the book is tailored around the architecture of a widely used Texas Instrument’s microcontroller, the MSP430. Instructor’s supplemental materials available through the book web site include solutions to selected problems and exercises and power point slides for lectures. The site also includes materials for students that include links to application examples and to sites elsewhere in the web with application notes, downloadab...

Genetic diversity in the C-terminus of merozoite surface protein 1 among Plasmodium knowlesi isolates from Selangor and Sabah Borneo, Malaysia.

Science.gov (United States)

Yap, Nan Jiun; Goh, Xiang Ting; Koehler, Anson V; William, Timothy; Yeo, Tsin Wen; Vythilingam, Indra; Gasser, Robin B; Lim, Yvonne A L

2017-10-01

Plasmodium knowlesi, a malaria parasite of macaques, has emerged as an important parasite of humans. Despite the significance of P. knowlesi malaria in parts of Southeast Asia, very little is known about the genetic variation in this parasite. Our aim here was to explore sequence variation in a molecule called the 42kDa merozoite surface protein-1 (MSP-1), which is found on the surface of blood stages of Plasmodium spp. and plays a key role in erythrocyte invasion. Several studies of P. falciparum have reported that the C-terminus (a 42kDa fragment) of merozoite surface protein-1 (MSP-1 42 ; consisting of MSP-1 19 and MSP-1 33 ) is a potential candidate for a malaria vaccine. However, to date, no study has yet investigated the sequence diversity of the gene encoding P. knowlesi MSP-1 42 (comprising Pk-msp-1 19 and Pk-msp-1 33 ) among isolates in Malaysia. The present study explored this aspect. Twelve P. knowlesi isolates were collected from patients from hospitals in Selangor and Sabah Borneo, Malaysia, between 2012 and 2014. The Pk-msp-1 42 gene was amplified by PCR and directly sequenced. Haplotype diversity (Hd) and nucleotide diversity (л) were studied among the isolates. There was relatively high genetic variation among P. knowlesi isolates; overall Hd and л were 1±0.034 and 0.01132±0.00124, respectively. A total of nine different haplotypes related to amino acid alterations at 13 positions, and the Pk-MSP-1 19 sequence was found to be more conserved than Pk-msp-1 33 . We have found evidence for negative selection in Pk-msp- 42 as well as the 33kDa and 19kDa fragments by comparing the rate of non-synonymous versus synonymous substitutions. Future investigations should study large numbers of samples from disparate geographical locations to critically assess whether this molecule might be a potential vaccine target for P. knowlesi. Copyright © 2017 Elsevier B.V. All rights reserved.

Effect of malaria transmission reduction by insecticide-treated bed nets (ITNs) on the genetic diversity of Plasmodium falciparum merozoite surface protein (MSP-1) and circumsporozoite (CSP) in western Kenya.

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Kariuki, Simon K; Njunge, James; Muia, Ann; Muluvi, Geofrey; Gatei, Wangeci; Ter Kuile, Feiko; Terlouw, Dianne J; Hawley, William A; Phillips-Howard, Penelope A; Nahlen, Bernard L; Lindblade, Kim A; Hamel, Mary J; Slutsker, Laurence; Shi, Ya Ping

2013-08-27

Although several studies have investigated the impact of reduced malaria transmission due to insecticide-treated bed nets (ITNs) on the patterns of morbidity and mortality, there is limited information on their effect on parasite diversity. Sequencing was used to investigate the effect of ITNs on polymorphisms in two genes encoding leading Plasmodium falciparum vaccine candidate antigens, the 19 kilodalton blood stage merozoite surface protein-1 (MSP-1(19kDa)) and the Th2R and Th3R T-cell epitopes of the pre-erythrocytic stage circumsporozoite protein (CSP) in a large community-based ITN trial site in western Kenya. The number and frequency of haplotypes as well as nucleotide and haplotype diversity were compared among parasites obtained from children diversity of > 0.7. No MSP-1(19kDa) 3D7 sequence-types were detected in 1996 and the frequency was less than 4% in 2001. The CSP Th2R and Th3R domains were highly polymorphic with a total of 26 and 14 haplotypes, respectively detected in 1996 and 34 and 13 haplotypes in 2001, with an overall haplotype diversity of > 0.9 and 0.75 respectively. The frequency of the most predominant Th2R and Th3R haplotypes was 14 and 36%, respectively. The frequency of Th2R and Th3R haplotypes corresponding to the 3D7 parasite strain was less than 4% at both time points. There was no significant difference in nucleotide and haplotype diversity in parasite isolates collected at both time points. High diversity in these two genes has been maintained overtime despite marked reductions in malaria transmission due to ITNs use. The frequency of 3D7 sequence-types was very low in this area. These findings provide information that could be useful in the design of future malaria vaccines for deployment in endemic areas with high ITN coverage and in interpretation of efficacy data for malaria vaccines based on 3D7 parasite strains.

Molecular detection of Dirofilaria immitis, Hepatozoon canis, Babesia spp., Anaplasma platys and Ehrlichia canis in dogs on Costa Rica.

Science.gov (United States)

Wei, Lanjing; Kelly, Patrick; Ackerson, Kate; El-Mahallawy, Heba S; Kaltenboeck, Bernhard; Wang, Chengming

2014-03-01

Although vector-borne diseases are important causes of morbidity and mortality in dogs in tropical areas, there is little information on these conditions in Costa Rica. In PCRs of blood from dogs in Costa Rica, we did not detect DNAs of Rickettsia (R.) felis and Coxiella (C.) burnetii but we did find evidence of infection with Dirofilaria (D.) immitis (9/40, 22.5%), Hepatozoon (H.) canis (15/40, 37.5%), Babesia spp. (10/40, 25%; 2 with B. gibsoni and 8 with B. vogeli), Anaplasma (A.) platys (3/40, 7.5%) and Ehrlichia (E.) canis (20/40, 50%). Nine dogs (22.5%) were free of any vector-borne pathogens while 14 (35%) were infected with a single pathogen, 11 (27.5%) with two, 4 (10%) with three, 1 (2.5%) with four, and 1 (2.5%) with five pathogens. Dogs in Costa Rica are commonly infected with vector-borne agents.

Anaplasma phagocytophilum increases the levels of histone modifying enzymes to inhibit cell apoptosis and facilitate pathogen infection in the tick vector Ixodes scapularis

Czech Academy of Sciences Publication Activity Database

Cabezas-Cruz, A.; Alberdi, P.; Ayllón, N.; Valdés, James J.; Pierce, R.; Villar, M.; de la Fuente, J.

2016-01-01

Roč. 11, č. 4 (2016), s. 303-319 ISSN 1559-2294 EU Projects: European Commission(XE) 278976 - ANTIGONE; European Commission(XE) 316304 - MODBIOLIN Institutional support: RVO:60077344 Keywords : Anaplasma * epigenetic s * histone modifying enzyme * histone * pathogen * tick Subject RIV: GJ - Animal Vermins ; Diseases, Veterinary Medicine Impact factor: 4.394, year: 2016

hnRNPA2B1 and hnRNPA1 mutations are rare in patients with "multisystem proteinopathy" and frontotemporal lobar degeneration phenotypes.

Science.gov (United States)

Le Ber, Isabelle; Van Bortel, Inge; Nicolas, Gael; Bouya-Ahmed, Kawtar; Camuzat, Agnès; Wallon, David; De Septenville, Anne; Latouche, Morwena; Lattante, Serena; Kabashi, Edor; Jornea, Ludmila; Hannequin, Didier; Brice, Alexis

2014-04-01

hnRNPA2B1 and hnRNPA1 mutations have been recently identified by exome sequencing in three families presenting with multisystem proteinopathy (MSP), a rare complex phenotype associating frontotemporal lobar degeneration (FTLD), Paget disease of bone (PDB), inclusion body myopathy (IBM), and amyotrophic lateral sclerosis (ALS). No study has evaluated the exact frequency of these genes in cohorts of MSP or FTD patients so far. We sequenced both genes in 17 patients with MSP phenotypes, and in 60 patients with FTLD and FTLD-ALS to test whether mutations could be implicated in the pathogenesis of these disorders. No disease-causing mutation was identified. We conclude that hnRNPA2B1 and hnRNPA1 mutations are rare in MSP and FTLD spectrum of diseases, although further investigations in larger populations are needed. Copyright © 2014 Elsevier Inc. All rights reserved.

A proline racemase based PCR for identification of Trypanosoma vivax in cattle blood.

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Regassa Fikru

Full Text Available A study was conducted to develop a Trypanosoma vivax (T. vivax specific PCR based on the T. vivax proline racemase (TvPRAC gene. Forward and reverse primers were designed that bind at 764-783 bp and 983-1002 bp of the gene. To assess its specificity, TvPRAC PCR was conducted on DNA extracted from different haemotropic pathogens: T. vivax from Nigeria, Ethiopia and Venezuela, T. congolense Savannah type, T. brucei brucei, T. evansi, T. equiperdum, T. theileri, Theileria parva, Anaplasma marginale, Babesia bovis and Babesia bigemina and from bovine, goat, mouse, camel and human blood. The analytical sensitivity of the TvPRAC PCR was compared with that of the ITS-1 PCR and the 18S PCR-RFLP on a dilution series of T. vivax DNA in water. The diagnostic performance of the three PCRs was compared on 411 Ethiopian bovine blood specimens collected in a former study. TvPRAC PCR proved to be fully specific for T. vivax, irrespective of its geographical origin. Its analytical sensitivity was lower than that of ITS-1 PCR. On these bovine specimens, TvPRAC PCR detected 8.3% T. vivax infections while ITS-1 PCR and 18S PCR-RFLP detected respectively 22.6 and 6.1% T. vivax infections. The study demonstrates that a proline racemase based PCR could be used, preferably in combination with ITS-1 PCR, as a species-specific diagnostic test for T. vivax infections worldwide.

Remodeling of tick cytoskeleton in response to infection with Anaplasma phagocytophilum.

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Cabezas-Cruz, Alejandro; Alberdi, Pilar; Valdes, James J; Villar, Margarita; de la Fuente, Jose

2017-06-01

The obligate intracellular pathogen Anaplasma phagocytophilum infects vertebrate and tick hosts. In this study, a genome-wide search for cytoskeleton components was performed in the tick vector, Ixodes scapularis . The available transcriptomics and proteomics data was then used to characterize the mRNA and protein levels of I. scapularis cytoskeleton components in response to A. phagocytophilum infection. The results showed that cytoskeleton components described in other model organisms were present in the I. scapularis genome. One type of intermediate filaments (lamin), a family of septins that was recently implicated in the cellular response to intracellular pathogens, and several members of motor proteins (kinesins and dyneins) that could be implicated in the cytoplasmic movements of A. phagocytophilum were found. The results showed that levels of tubulin, actin, septin, actin-related proteins and motor proteins were affected by A. phagocytophilum , probably to facilitate infection in I. scapularis . Functional studies demonstrated a role for selected cytoskeleton components in pathogen infection. These results provided a more comprehensive view of the cytoskeletal components involved in the response to A. phagocytophilum infection in ticks.

Clinical and molecular features of one case of human infection with Anaplasma phagocytophilum from Podlaskie Province in eastern Poland

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Renata Welc-Falęciak

2015-09-01

Full Text Available The article focuses on the clinical and laboratory diagnosis of human granulocytic anaplasmosis (HGA caused by Anaplasma phagocytophilum infection in one of 28 patients (3.6%; n=1/28 tested samples with early Lyme borreliosis. The clinical and laboratory results of a 42-year-old patient fulfilled criteria of confirm anaplasmosis and suggest an acute stage of illness. The described case provides strong presumptive evidence that infection in this patient was acquired with a pathogenic strain of A. phagocytophilum through a tick bite. A positive DNA with PCR for A. phagocytophilum infection was sequenced and analyzed phylogenetically. Physicians should consider the possibility of anaplasmosis in patients with early Lyme borreliosis, and A. phagocytophilum should be considered as a differential diagnosis in all patients from an endemic region of potential high risk factors for tick-borne diseases.

Human Exposure to Anaplasma phagocytophilum in Two Cities of Northwestern Morocco

Science.gov (United States)

Elhamiani Khatat, Sarah; Sahibi, Hamid; Hing, Mony; Alaoui Moustain, Ismail; El Amri, Hamid; Benajiba, Mohammed; Kachani, Malika; Duchateau, Luc; Daminet, Sylvie

2016-01-01

Anaplasma phagocytophilum is an emerging tick-borne zoonosis with extensive increased interest. Epidemiological data are available in several regions of the USA, Europe and Asia in contrast to other parts of the world such as North Africa. Blood samples of 261 healthy individuals divided in two groups i.e., dog handlers and blood donors were analysed. Indirect immunofluorescent assay using a commercial kit was performed to detect specific A. phagocytophilum IgG. Two dilutions were used to assess the prevalence of seroreactive samples. Demographic variables were assessed as potential risk factors using exact logistic regression. Seropositivity rates reached 37% and 27% in dog handlers and 36% and 22% in blood donors. No statistically significant differences were found in the prevalence rates between the two groups. Analysis of risk factors such as gender, age groups, outdoor activities, self-reported previous exposure to ticks, or contact with domestic animals (dogs, cats, ruminants and horses) did not shown any significant difference. A. phagocytophilum exposure was common in both high-risk population and blood donors in Morocco. PMID:27532208

Superinfection occurs in Anaplasma phagocytophilum infected sheep irrespective of infection phase and protection status

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Bergström Karin

2009-10-01

Full Text Available Abstract Background Anaplasma phagocytophilum infection in domestic ruminants is widespread in the coastal areas of southern Norway. The bacteria may persist in mammalian hosts. Several genetic variants of A. phagocytophilum exist. In the present study, we investigate whether superinfection occurs in the acute and persistent phase of the infection. Methods Five-month-old lambs of the Norwegian Dala breed were experimentally infected with two 16S rRNA gene variants of A. phagocytophilum, i.e. A. phagocytophilum variant 1 (GenBank accession number M73220 and variant 2 (GenBank acc. no. AF336220. Eighteen lambs were used, two lambs in each group. Eight groups were experimentally inoculated with either variant 1 or 2 on day 0. Six of these groups were then challenged with the other variant on either days 7, 42 or 84, respectively. One group was left uninfected. The occurrence of A. phagocytophilum in blood samples was determined using semi-nested PCR analysis and gene sequencing. Specific antibodies were measured by an indirect immunofluorescence antibody assay (IFA. Results A. phagocytophilum variant 1 and 2 differed significantly with regards to clinical reaction and cross-immunity in infected lambs. Both variants were found in the blood after challenge. However, variant 1 was detected most frequently. Conclusion The present experiment indicates that superinfection of different genotypes occurs during the acute as well as the persistent phase of an A. phagocytophilum infection, even in lambs protected against the challenged infection.

Molecular and serological detection of Theileria equi, Babesia caballi and Anaplasma phagocytophilum in horses and ticks in Maranhão, Brazil

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Rita de Maria Seabra Nogueira

Full Text Available ABSTRACT: Equine piroplasmosis is a tick-borne disease caused by the intraeytrhocytic protozoans Babesia caballi and Theileria equi. It has been reported as a main equine parasitic disease. In addition, Anaplasma phagocytophilum, the causative agent of granulocytic ehrlichiosis, causes a seasonal disease in horses. Both diseases, can be detrimental to animal health. In this sense, blood samples and ticks were collected from 97 horses raised in the microregion of Baixada Maranhense, Maranhão State, Brazil. Serum samples were subjected to Indirect Fluorescence Antibody Test (IFAT and blood samples and ticks to Polymerase Chain Reaction (PCR to evaluate the infection by Theileria equi, Babesia caballi and Anaplasma phagocytophilum. The overall seroprevalence was 38.14%, 18.55% and 11.34% for T. equi, B. caballi and A. phagocytophilum, respectively. The results of PCR from blood samples showed 13.40% and 3.09% positive samples to T. equi and B. caballi, respectively. A total of 170 tick specimens were collected and identified as Dermacentor nitens, Amblyomma cajennense sensu lato and Rhipicephalus (Boophilus microplus. It was detected 2.35% (4/170 and 0.59% (1/170 positive tick samples by PCR for T. equi and B. caballi, respectively. All samples were negative to A. phagocytophilum. No statically difference (p>0.05 was observed when gender, age, use of ectoparasiticide and tick presence were analyzed. A BLASTn analysis of the sequenced samples indicated 97 to 100% similarity with T. equi 18S rRNA gene sequences in GenBank and 98 to 100% with B. caballi. Genetic analysis classified the obtained sequences as T. equi and B. caballi cluster, respectively. It can be concluded that these pathogens occur and are circulating in the studied area.

Temporal stability of naturally acquired immunity to Merozoite Surface Protein-1 in Kenyan Adults

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Crabb Brendan S

2009-07-01

Full Text Available Abstract Background Naturally acquired immunity to blood-stage Plasmodium falciparum infection develops with age and after repeated infections. In order to identify immune surrogates that can inform vaccine trials conducted in malaria endemic populations and to better understand the basis of naturally acquired immunity it is important to appreciate the temporal stability of cellular and humoral immune responses to malaria antigens. Methods Blood samples from 16 adults living in a malaria holoendemic region of western Kenya were obtained at six time points over the course of 9 months. T cell immunity to the 42 kDa C-terminal fragment of Merozoite Surface Protein-1 (MSP-142 was determined by IFN-γ ELISPOT. Antibodies to the 42 kDa and 19 kDa C-terminal fragments of MSP-1 were determined by serology and by functional assays that measure MSP-119 invasion inhibition antibodies (IIA to the E-TSR (3D7 allele and growth inhibitory activity (GIA. The haplotype of MSP-119 alleles circulating in the population was determined by PCR. The kappa test of agreement was used to determine stability of immunity over the specified time intervals of 3 weeks, 6 weeks, 6 months, and 9 months. Results MSP-1 IgG antibodies determined by serology were most consistent over time, followed by MSP-1 specific T cell IFN-γ responses and GIA. MSP-119 IIA showed the least stability over time. However, the level of MSP-119 specific IIA correlated with relatively higher rainfall and higher prevalence of P. falciparum infection with the MSP-119 E-TSR haplotype. Conclusion Variation in the stability of cellular and humoral immune responses to P. falciparum blood stage antigens needs to be considered when interpreting the significance of these measurements as immune endpoints in residents of malaria endemic regions.

Poly(I:C) adjuvant strongly enhances parasite-inhibitory antibodies and Th1 response against Plasmodium falciparum merozoite surface protein-1 (42-kDa fragment) in BALB/c mice.

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Mehrizi, Akram Abouie; Rezvani, Niloufar; Zakeri, Sedigheh; Gholami, Atefeh; Babaeekhou, Laleh

2018-04-01

Malaria vaccine development has been confronted with various challenges such as poor immunogenicity of malaria vaccine candidate antigens, which is considered as the main challenge. However, this problem can be managed using appropriate formulations of antigens and adjuvants. Poly(I:C) is a potent Th1 inducer and a human compatible adjuvant capable of stimulating both B- and T-cell immunity. Plasmodium falciparum merozoite surface protein 1 42 (PfMSP-1 42 ) is a promising vaccine candidate for blood stage of malaria that has faced several difficulties in clinical trials, mainly due to improper adjuvants. Therefore, in the current study, poly(I:C), as a potent Th1 inducer adjuvant, was evaluated to improve the immunogenicity of recombinant PfMSP-1 42 , when compared to CFA/IFA, as reference adjuvant. Poly(I:C) produced high level and titers of anti-PfMSP-1 42 IgG antibodies in which was comparable to CFA/IFA adjuvant. In addition, PfMSP-1 42 formulated with poly(I:C) elicited a higher ratio of IFN-γ/IL-4 (23.9) and IgG2a/IgG1 (3.77) with more persistent, higher avidity, and titer of IgG2a relative to CFA/IFA, indicating a potent Th1 immune response. Poly(I:C) could also help to induce anti-PfMSP-1 42 antibodies with higher growth-inhibitory activity than CFA/IFA. Altogether, the results of the current study demonstrated that poly(I:C) is a potent adjuvant that can be appropriate for being used in PfMSP-1 42 -based vaccine formulations.

Methylation profiling of SOCS1, SOCS2, SOCS3, CISH and SHP1 in Philadelphia-negative myeloproliferative neoplasm.

Science.gov (United States)

Zhang, Min Yue; Fung, Tsz Kin; Chen, Fang Yuan; Chim, Chor Sang

2013-10-01

Janus kinase-signal transducer and activator of transcription (JAK/STAT) signalling, pivotal in Philadelphia-negative (Ph-ve) myeloproliferative neoplasm (MPN), is negatively regulated by molecules including SOCSs, CISH and SHP1. SOCS1, SOCS2 and SOCS3 methylation have been studied in MPN with discordant results. Herein, we studied the methylation status of SOCS1, SOCS2 and SOCS3, CISH and SHP1 by methylation-specific polymerase chain reaction (MSP) in cell lines and 45 diagnostic marrow samples of Ph-ve MPN. Moreover, we attempted to explain the discordance of methylation frequency by mapping the studied MSP primers to the respective genes. Methylation was detected in normal controls using SOCS2 MSP primers in the 3'translated exonic sequence, but not primers around the transcription start site in the 5' untranslated regions (5'UTR). SOCS1, SOCS2, SOCS3 and CISH were completely unmethylated in primary MPN samples and cell lines. In contrast, methylation of SHP1 was detected in 8.9% primary marrow samples. Moreover, SHP1 was completely methylated in K562 cell line, leading to reversible SHP1 silencing. A review of methylation studies of SOCS1 and SOCS3 showed that spuriously high rates of SOCS methylation had been reported using MSP primers targeting CpG sites in the 3'translated exonic sequence, which is also methylated in normal controls. However, using MSP primers localized to the 5'UTR, methylation of SOCS1, SOCS2 and SOCS3 is infrequent across all studies. In summary, methylation of SOCS1, SOCS2, SOCS3 and CISH is infrequent in Ph-ve MPN. Appropriate MSP primers are important for accurate estimation of the methylation frequency. The role of SHP1 methylation in the pathogenesis of MPN warrants further investigation. © 2013 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.

Direct Detection of Unnatural DNA Nucleotides dNaM and d5SICS using the MspA Nanopore.

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Jonathan M Craig

Full Text Available Malyshev et al. showed that the four-letter genetic code within a living organism could be expanded to include the unnatural DNA bases dNaM and d5SICS. However, verification and detection of these unnatural bases in DNA requires new sequencing techniques. Here we provide proof of concept detection of dNaM and d5SICS in DNA oligomers via nanopore sequencing using the nanopore MspA. We find that both phi29 DNA polymerase and Hel308 helicase are capable of controlling the motion of DNA containing dNaM and d5SICS through the pore and that single reads are sufficient to detect the presence and location of dNaM and d5SICS within single molecules.

N-Terminal Plasmodium vivax Merozoite Surface Protein-1, a Potential Subunit for Malaria Vivax Vaccine

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Fernanda G. Versiani

2013-01-01

Full Text Available The human malaria is widely distributed in the Middle East, Asia, the western Pacific, and Central and South America. Plasmodium vivax started to have the attention of many researchers since it is causing diseases to millions of people and several reports of severe malaria cases have been noticed in the last few years. The lack of in vitro cultures for P. vivax represents a major delay in developing a functional malaria vaccine. One of the major candidates to antimalarial vaccine is the merozoite surface protein-1 (MSP1, which is expressed abundantly on the merozoite surface and capable of activating the host protective immunity. Studies have shown that MSP-1 possesses highly immunogenic fragments, capable of generating immune response and protection in natural infection in endemic regions. This paper shows humoral immune response to different proteins of PvMSP1 and the statement of N-terminal to be added to the list of potential candidates for malaria vivax vaccine.

Longitudinal field study on bovine Babesia spp. and Anaplasma phagocytophilum infections during a grazing season in Belgium.

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Lempereur, Laetitia; Lebrun, Maude; Cuvelier, Pascale; Sépult, Géraldine; Caron, Yannick; Saegerman, Claude; Shiels, Brian; Losson, Bertrand

2012-04-01

Anaplasmosis and babesiosis are major tick-borne diseases with a high economic impact but are also a public health concern. Blood samples collected in the spring, summer, and autumn of 2010 from 65 cows in seven different farms in Belgium were monitored with an indirect immunofluorescence antibody test to assess seroprevalence against these pathogens. Seroprevalences to Babesia spp. were measured as 10.7%, 20%, and 12.3% in spring, summer, and autumn, respectively, whereas seroprevalences to Anaplasma phagocytophilum were 30.8%, 77%, and 56.9%, respectively. A total of 805 Ixodes ricinus ticks were collected at the same time from both cattle (feeding ticks) and grazed pastures (questing ticks). The infection level of ticks, assessed by PCR assay, for Babesia spp. DNA was 14.6% and 7.9% in feeding and questing ticks, respectively, whereas 21.7% and 3% of feeding and questing ticks were found be positive for A. phagocytophilum cDNA. Fifty-five PCR-positive samples were identified by sequencing as Babesia sp. EU1, of which five from feeding ticks were positive for both A. phagocytophilum and Babesia sp. EU1. The high density of wild cervids in the study area could explain these observations, as deer are considered to be the main hosts for adults of I. ricinus. However, the absence of Babesia divergens both in feeding and questing ticks is surprising, as the study area is known to be endemic for cattle babesiosis. Increasing cervid populations and comorbidity could play an import role in the epidemiology of these tick-borne diseases.

OCORRÊNCIA DE Anaplasma bovis (Donatien & Lestoquard, 1936, Dumler et al. 2001 NA REGIÃO DE BOM JESUS DO ITABAPOANA, RJ

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Orlando Augusto Melo Junior

2010-12-01

Full Text Available In the present study, 400 capillary blood smears collected from the ear and the tail of 200 bovines were microscopically examinedfor the presence of hemoparasites. The diagnosis was based on the direct research, semonstrating the etiologic agent inclusions. Eleven bovines (5.5% were considered infected by Anaplasma bovis, confirming the presence of this hemoparasite in the city of Bom Jesus do Itabapoana, the first city of the northwesten region of the state of Rio de Janeiro to register such occurrence.

Tristeza parasitária bovina na região sul do Rio Grande do Sul: estudo retrospectivo de 1978-2005 Tick fever in southern Brazil: a retrospective study of 1978-2005

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Milton Begeres de Almeida

2006-12-01

Full Text Available Foi realizado um estudo retrospectivo dos casos de tristeza parasitária bovina (TPB ocorridos no sul do Rio Grande do Sul, área de influência do Laboratório Regional de Diagnóstico (LRD da Faculdade de Veterinária da Universidade Federal de Pelotas entre 1978 e 2005. De um total de 4.884 materiais de bovinos provenientes de necropsias realizadas e órgãos ou sangue enviados ao laboratório 231 (4,7% tiveram o diagnóstico de TPB. Desses 231 surtos foram resgatados os dados de 221 diagnósticos dos quais 91 (41,1% foram causados por Babesia bovis, 11 (4,9% por Babesia bigemina, e 65 (29,41% por Anaplasma marginale. Em outros 33 (14,93% surtos de babesiose não foi informada a espécie de Babesia e em 21 (9,5% surtos foi detectada infecção mista por Babesia sp e A. marginale. Os índices gerais médios de morbidade, mortalidade e letalidade, resgatados em 149 dos 221 surtos da doença, foram de 11,17%, 6,81% e 70,04%, respectivamente. Verificou-se que, na região estudada, a maioria dos surtos ocorre durante os meses de verão e outono, e que os animais com um a três anos de idade são os mais afetados. Os sinais clínicos nos surtos caracterizaram-se por apatia, orelhas caídas, debilidade, febre, anorexia e emagrecimento. Os valores de hematócrito eram baixos. Hemoglobinúria foi frequentemente observada nos casos de babesiose. Sinais neurológicos estavam presentes nos casos de babesiose por B. bovis e se caracterizaram por transtornos da locomoção, tremores musculares, agressividade e quedas com movimentos de pedalagem. As lesões macroscópicas principais relatadas nos casos de babesiose foram esplenomegalia, hepatomegalia, fígado amarelo, hemoglobinúria, icterícia, hemorragias cardíacas e bile espessa. Congestão do córtex cerebral foi relatada nos casos de babesiose por B. bovis. Nesta região, com população de bovinos de aproximadamente 2.630.000 cabeças as perdas anuais por morte de bovinos pela enfermidade podem ser

Genetic diversity of Plasmodium Vivax revealed by the merozoite surface protein-1 icb5-6 fragment.

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Ruan, Wei; Zhang, Ling-Ling; Feng, Yan; Zhang, Xuan; Chen, Hua-Liang; Lu, Qiao-Yi; Yao, Li-Nong; Hu, Wei

2017-06-05

Plasmodium vivax remains a potential cause of morbidity and mortality for people living in its endemic areas. Understanding the genetic diversity of P. vivax from different regions is valuable for studying population dynamics and tracing the origins of parasites. The PvMSP-1 gene is highly polymorphic and has been used as a marker in many P. vivax population studies. The aim of this study was to investigate the genetic diversity of the PvMSP-1 gene icb5-6 fragment and to provide more genetic polymorphism data for further studies on P. vivax population structure and tracking of the origin of clinical cases. Nested PCR and sequencing of the PvMSP-1 icb5-6 marker were performed to obtain the nucleotide sequences of 95 P. vivax isolates collected from Zhejiang province, China. To investigate the genetic diversity of PvMSP-1, the 95 nucleotide sequences of the PvMSP-1 icb5-6 fragment were genotyped and analyzed using DnaSP v5, MEGA software. The 95 P. vivax isolates collected from Zhejiang province were either indigenous cases or imported cases from different regions around the world. A total of 95 sequences ranging from 390 to 460 bp were obtained. The 95 sequences were genotyped into four allele-types (Sal I, Belem, R-III and R-IV) and 17 unique haplotypes. R-III and Sal I were the predominant allele-types. The haplotype diversity (Hd) and nucleotide diversity (Pi) were estimated to be 0.729 and 0.062, indicating that the PvMSP-1 icb5-6 fragment had the highest level of polymorphism due to frequent recombination processes and single nucleotide polymorphism. The values of dN/dS and Tajima's D both suggested neutral selection for the PvMSP-1icb5-6 fragment. In addition, a rare recombinant style of R-IV type was identified. This study presented high genetic diversity in the PvMSP-1 marker among P. vivax strains from around the world. The genetic data is valuable for expanding the polymorphism information on P. vivax, which could be helpful for further study on

Genetic Diversity and Natural Selection in 42 kDa Region of Plasmodium vivax Merozoite Surface Protein-1 from China-Myanmar Endemic Border.

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Zhou, Xia; Tambo, Ernest; Su, Jing; Fang, Qiang; Ruan, Wei; Chen, Jun-Hu; Yin, Ming-Bo; Zhou, Xiao-Nong

2017-10-01

Plasmodium vivax merozoite surface protein-1 (PvMSP1) gene codes for a major malaria vaccine candidate antigen. However, its polymorphic nature represents an obstacle to the design of a protective vaccine. In this study, we analyzed the genetic polymorphism and natural selection of the C-terminal 42 kDa fragment within PvMSP1 gene (Pv MSP142) from 77 P. vivax isolates, collected from imported cases of China-Myanmar border (CMB) areas in Yunnan province and the inland cases from Anhui, Yunnan, and Zhejiang province in China during 2009-2012. Totally, 41 haplotypes were identified and 30 of them were new haplotypes. The differences between the rates of non-synonymous and synonymous mutations suggest that PvMSP142 has evolved under natural selection, and a high selective pressure preferentially acted on regions identified of PvMSP133. Our results also demonstrated that PvMSP142 of P. vivax isolates collected on China-Myanmar border areas display higher genetic polymorphisms than those collected from inland of China. Such results have significant implications for understanding the dynamic of the P. vivax population and may be useful information towards China malaria elimination campaign strategies.

Genetic diversity in the merozoite surface protein 1 and 2 genes of Plasmodium falciparum from the Artibonite Valley of Haiti.

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Londono-Renteria, Berlin; Eisele, Thomas P; Keating, Joseph; Bennett, Adam; Krogstad, Donald J

2012-01-01

Describing genetic diversity of the Plasmodium falciparum parasite provides important information about the local epidemiology of malaria. In this study, we examined the genetic diversity of P. falciparum isolates from the Artibonite Valley in Haiti using the allelic families of merozoite surface protein 1 and 2 genes (msp-1 and msp-2). The majority of study subjects infected with P. falciparum had a single parasite genotype (56% for msp-1 and 69% for msp-2: n=79); 9 distinct msp-1 genotypes were identified by size differences on agarose gels. K1 was the most polymorphic allelic family with 5 genotypes (amplicons from 100 to 300 base pairs [bp]); RO33 was the least polymorphic, with a single genotype (120-bp). Although both msp-2 alleles (3D7/IC1, FC27) had similar number of genotypes (n=4), 3D7/IC1 was more frequent (85% vs. 26%). All samples were screened for the presence of the K76T mutation on the P. falciparum chloroquine resistance transporter (pfcrt) gene with 10 of 79 samples positive. Of the 2 (out of 10) samples from individuals follow-up for 21 days, P. falciparum parasites were present through day 7 after treatment with chloroquine. No parasites were found on day 21. Our results suggest that the level of genetic diversity is low in this area of Haiti, which is consistent with an area of low transmission. Copyright © 2011 Elsevier B.V. All rights reserved.

The infection of questing Dermacentor reticulatus ticks with Babesia canis and Anaplasma phagocytophilum in the Chernobyl exclusion zone.

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Karbowiak, Grzegorz; Vichová, Bronislavá; Slivinska, Kateryna; Werszko, Joanna; Didyk, Julia; Peťko, Branislav; Stanko, Michal; Akimov, Igor

2014-08-29

Tick occurrence was studied in the Chernobyl exclusion zone (CEZ) during the August-October 2009-2012. Dermacentor reticulatus ticks were collected using the flagging method and then screened for infection with Anaplasma phagocytophilum and Babesia canis by a PCR method incorporating specific primers and sequence analysis. The prevalence of infection with B. canis canis and A. phagocytophilum was found to be 3.41% and 25.36%, respectively. The results present the first evidence of B. canis canis and A. phagocytophilum in questing D. reticulatus ticks from the Chernobyl exclusion zone. They also reveal the presence of tick-borne disease foci in areas with no human activity, and confirm that they can be maintained in areas after a nuclear disaster with radioactive contamination. Copyright © 2014 Elsevier B.V. All rights reserved.

Intraoperative bleeding in dogs from Grenada seroreactive to Anaplasma platys and Ehrlichia canis.

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Lanza-Perea, M; Zieger, U; Qurollo, B A; Hegarty, B C; Pultorak, E L; Kumthekar, S; Bruhl-Day, R; Breitschwerdt, E B

2014-01-01

Frequent exposure of Grenadian dogs to Rhipicephalus sanguineus results in Anaplasma platys, and Ehrlichia canis seroreactivity. During elective surgeries, substantial intraoperative hemorrhage occurs in some seroreactive dogs. To assess hemostatic parameters and bleeding tendencies as well as prevalence of PCR positivity in apparently healthy A. platys and E. canis seroreactive and seronegative free-roaming dogs from Grenada. Forty-seven elective surgery dogs allocated to 4 groups: Seronegative control (n = 12), A. platys (n = 10), E. canis (n = 14) and A. platys, and E. canis (n = 11) seroreactive. Preoperatively, hemostasis was assessed by platelet count, prothrombin time, activated partial thromboplastin time, and buccal mucosal bleeding time. Intra- and postoperative bleeding scores were subjectively assigned. Blood, spleen, bone marrow, and lymph node aspirates were tested by PCR. Bleeding scores in dogs coseroreactive for A. platys and E. canis were higher (P = .015) than those of seronegative dogs. A. platys DNA was amplified from 7/21 (33%) A. platys seroreactive dogs and from 1 E. canis seroreactive dog; E. canis DNA was amplified from 21/25 (84%) E. canis seroreactive dogs. E. canis DNA was amplified most often from blood, whereas A. platys DNA was amplified most often from bone marrow. Apparently healthy, free-roaming dogs coseropositive for A. platys and E. canis may have increased intraoperative bleeding tendencies despite normal hemostatic parameters. Future investigations should explore the potential for vascular injury as a cause for bleeding in these dogs. Improved tick control is needed for dogs in Grenada. Copyright © 2014 by the American College of Veterinary Internal Medicine.

Los complejos Chaperonina(MSP63inducen anticuerpos de reacciones cruzadas, bactericidas y opsonofagocítica.

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Juan Marzoa

2009-08-01

Full Text Available Alteration of the native structure of antigens can lead to the loss of protective epitopes. Our previous results showed that separation of the meningococcal outer membrane proteins in native conditions revealed the existence of protein complexes that could be relevant for the development of new vaccine formulations. The aim of this work was to analyse the immunogenic characteristics of a highly conserved 700 kDa chaperonin complex (CxChap detected and purified by using high resolution clear native electrophoresis. Analysis of the anti-CxChap serum by Western-blotting revealed the presence of antibodies against the MSP63 but also against the macrophage infectivity potentiator-like protein (MIP, which is coopurified with the chaperonin complex. Antibodies raised by immunisation with CxChap chaperonin complex show bactericidal and opsonophagocytic activity.

Plasmodium falciparum merozoite surface protein 1 - Glycosylation and localization to low-density, detergent-resistant membranes in the parasitized erythrocyte

DEFF Research Database (Denmark)

Hoessli, D.C.; Poincelet, M.; Gupta, Ramneek

2003-01-01

In addition to the major carbohydrate moieties of the glycosylphosphatidylinositol (GPI) anchor, we report that Plasmodium falciparum merozoite surface protein 1 (MSP-1) bears O-GlcNAc modifications predominantly in beta-anomeric configuration, in both the C- and N-terminal portions of the protein....... Subcellular fractionation of parasitized erythrocytes in the late trophozoite/schizont stage reveals that GPI-anchored C-terminal fragments of MSP-1 are recovered in Triton X-100 resistant, low-density membrane fractions. Our results suggest that O -GlcNAc-modified MSP-1 N-terminal fragments tend to localize...... within the parasitophorous vacuolar membrane while GPI-anchored MSP-1 C-terminal fragments associate with low-density, Triton X-100 resistant membrane domains (rafts), redistribute in the parasitized erythrocyte and are eventually shed as membrane vesicles that also contain the endogenous, GPI-linked CD...

The Prostaglandin E2-EP3 Receptor Axis Regulates Anaplasma phagocytophilum-Mediated NLRC4 Inflammasome Activation.

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Xiaowei Wang

2016-08-01

Full Text Available Rickettsial agents are sensed by pattern recognition receptors but lack pathogen-associated molecular patterns commonly observed in facultative intracellular bacteria. Due to these molecular features, the order Rickettsiales can be used to uncover broader principles of bacterial immunity. Here, we used the bacterium Anaplasma phagocytophilum, the agent of human granulocytic anaplasmosis, to reveal a novel microbial surveillance system. Mechanistically, we discovered that upon A. phagocytophilum infection, cytosolic phospholipase A2 cleaves arachidonic acid from phospholipids, which is converted to the eicosanoid prostaglandin E2 (PGE2 via cyclooxygenase 2 (COX2 and the membrane associated prostaglandin E synthase-1 (mPGES-1. PGE2-EP3 receptor signaling leads to activation of the NLRC4 inflammasome and secretion of interleukin (IL-1β and IL-18. Importantly, the receptor-interacting serine/threonine-protein kinase 2 (RIPK2 was identified as a major regulator of the immune response against A. phagocytophilum. Accordingly, mice lacking COX2 were more susceptible to A. phagocytophilum, had a defect in IL-18 secretion and exhibited splenomegaly and damage to the splenic architecture. Remarkably, Salmonella-induced NLRC4 inflammasome activation was not affected by either chemical inhibition or genetic ablation of genes associated with PGE2 biosynthesis and signaling. This divergence in immune circuitry was due to reduced levels of the PGE2-EP3 receptor during Salmonella infection when compared to A. phagocytophilum. Collectively, we reveal the existence of a functionally distinct NLRC4 inflammasome illustrated by the rickettsial agent A. phagocytophilum.

CYP1A1 Genetic Polymorphisms in Ecuador, South America

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César Paz-y-Miño

2005-01-01

Full Text Available A total of 108 individuals from the Ecuadorian population from rural and urban places were analyzed for two CYP1A1 gene polymorphisms. The frequency of the val allele at codon 462 was 0.50, while the frequency of the Msp I restriction site, m2 allele at the T6235C position was 0.70. These polymorphisms in Ecuador have higher frequencies if we compare with others around the world, with the exception of some South American population in Brazil and Chile.

Differential patterns of human immunoglobulin G subclass responses to distinct regions of a single protein, the merozoite surface protein 1 of Plasmodium falciparum

DEFF Research Database (Denmark)

Cavanagh, D R; Dobaño, C; Elhassan, I M

2001-01-01

Comparisons of immunoglobulin G (IgG) subclass responses to the major polymorphic region and to a conserved region of MSP-1 in three cohorts of African villagers exposed to Plasmodium falciparum revealed that responses to Block 2 are predominantly IgG3 whereas antibodies to MSP-1(19) are mainly IgG......1. The striking dominance of IgG3 to Block 2 may explain the short duration of this response and also the requirement for continuous stimulation by malaria infection to maintain clinical immunity....

Mental stress and psychosocial factors at work in relation to multiple-site musculoskeletal pain: a longitudinal study of kitchen workers.

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Haukka, Eija; Leino-Arjas, Päivi; Ojajärvi, Anneli; Takala, Esa-Pekka; Viikari-Juntura, Eira; Riihimäki, Hilkka

2011-04-01

Among 385 female kitchen workers, we examined (1) whether mental stress and psychosocial factors at work (job control, skill discretion, supervisor support, co-worker relationships, and hurry) predict multiple-site musculoskeletal pain (MSP; defined as pain at ≥ 3 of seven sites) and (2) reversedly, whether MSP predicts these psychosocial factors. Data were collected by questionnaire at 3-month intervals during 2 years. Trajectory analysis was applied. Four trajectories of MSP prevalence emerged: Low, Descending, Ascending, and High. For the psychosocial factors, a two-trajectory model (Ascending or High vs. Low) yielded the best fit. In logistic regression analysis, with the Low MSP trajectory as reference, poor co-worker relationships (odds ratio [OR] 3.9), mental stress (3.1) and hurry (2.1) at baseline predicted belonging to the High MSP trajectory. Also MSP at baseline predicted the trajectories (Ascending vs. Low) of low job control (2.2) and mental stress (3.2). Adverse changes in most psychosocial factors were associated with belonging to the High (ORs between 2.3 and 8.6) and Ascending (2.7-5.5) MSP trajectories. In generalized estimating equations, time-lagged by 3 months, all psychosocial factors but two predicted MSP (1.4-2.1), allowing, e.g. for MSP at baseline, and vice versa, MSP predicted low job control, low supervisor support, and mental stress (1.4-2.0), after adjustment for e.g. the relevant psychosocial factor at baseline. In conclusion, we found that several psychosocial factors predicted MSP and that MSP predicted several psychosocial factors. The results suggest a cumulative process in which adverse psychosocial factors and MSP influence each other. Copyright © 2010 European Federation of International Association for the Study of Pain Chapters. Published by Elsevier Ltd. All rights reserved.

Anaplasma phagocytophilum-Related Defects in CD8, NKT, and NK Lymphocyte Cytotoxicity

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Diana G. Scorpio

2018-04-01

Full Text Available Human granulocytic anaplasmosis, caused by the tick-transmitted Anaplasma phagocytophilum, is not controlled by innate immunity, and induces a proinflammatory disease state with innate immune cell activation. In A. phagocytophilum murine infection models, hepatic injury occurs with production of IFNγ thought to be derived from NK, NKT cells, and CD8 T lymphocytes. Specific A. phagocytophilum ligands that drive inflammation and disease are not known, but suggest a clinical and pathophysiologic basis strikingly like macrophage activation syndrome (MAS and hemophagocytic syndrome (HPS. We studied in vivo responses of NK, NKT, and CD8 T lymphocytes from infected animals for correlates of lymphocyte-mediated cytotoxicity and examined in vitro interactions with A. phagocytophilum-loaded antigen-presenting cells (APCs. Murine splenocytes were examined and found deficient in cytotoxicity as determined by CD107a expression in vitro for specific CTL effector subsets as determined by flow cytometry. Moreover, A. phagocytophilum-loaded APCs did not lead to IFNγ production among CTLs in vitro. These findings support the concept of impaired cytotoxicity with A. phagocytophilum presentation by APCs that express MHC class I and that interact with innate and adaptive immune cells with or after infection. The findings strengthen the concept of an enhanced proinflammatory phenotype, such as MAS and HPS disease states as the basis of disease and severity with A. phagocytophilum infection, and perhaps by other obligate intracellular bacteria.

Prevalence and Significance of Haemoparasitic Infections of Cattle in North- Central, Nigeria

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J.Kamani.

Full Text Available The prevalence and significance of hemoparasites of cattle from north-central Nigeria was determined using diagnostic records from Parasitology Division, National Veterinary Research Institute (NVRI Vom, from May 2006 to April 2008. A total of 637 blood samples from cattle from four states (Plateau, Bauchi, Nasarawa and Kaduna of Nigeria in anticoagulant were submitted to the laboratory for parasitological diagnosis. Giemsa stained thin blood smears were examined for hemoparasites. Packed cell volume (PCV for each sample was determined and Hematocrit centrifuge technique (HCT was used to determine the presence of motile parasites. An overall prevalence of 25.7% was recorded for all samples examined. Babesia bigemina and B.bovis accounted for 16.0%, followed by Theileria mutans (3.1%, Trypanosoma spp (T.vivax and T. congolense (2.8%, Anaplasma marginale (1.9%, Microfilaria (1.4%. The hemoparasites identified alone or in combination with others had a significant (P<0.05 effect on the mean PCV of infected animals. Similarly, hemoparasites infection in young animals as well as during the dry season resulted in significant (P<0.05 reduction of PCV values. The result of this study shows these hemoparasites are endemic in cattle in the study area which may result in serious disease conditions when such animals are subjected to stressful condition. [Veterinary World 2010; 3(10.000: 445-448

Responsabilidad Social Matriz de indicadores sustentables que refleja el impacto de la forestación en tierras marginales del Partido de Bahía Blanca, Argentina

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Regina del Carmen Durán Durán

2009-01-01

Full Text Available Actualmente, se han empezado a considerar otras obligaciones que tienen las empresas para con su comunidad; se trata de aspectos vinculados con las dimensiones sociales y ambientales. Desde este contexto, nuestro trabajo tiene como punto de partida el concepto de responsabilidad social desde la noción de sustentabilidad. El objetivo es evaluar la forestación como instrumento para mejorar la sostenibilidad de los sistemas productivos tradicionales en tierras marginales del Partido de Bahía Blanca, mitigando los impactos del desarrollo no responsable de la producción primaria. Para ello, se han identificado técnicas de manejo generadoras de externalidades positivas en otras actividades agropecuarias complementarias, donde existen limitantes edafológicas, apoyándose en las tres dimensiones del concepto de sustentabilidad: ecológica, económica, sociopolítica y cultural. De igual forma, se diseñó una matriz de evaluación y ponderación de impactos, que contempla indicadores cuantitativos y cualitativos, de cuya aplicación surge la forestación como una alternativa sustentable

The prevalence of Anaplasma phagocytophilum in questing Ixodes ricinus ticks in SW Poland.

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Kiewra, Dorota; Zaleśny, Grzegorz; Czułowska, Aleksandra

2014-01-01

Ticks constitute important vectors of human and animal pathogens. Besides the Lyme borreliosis and tick-borne encephalitis, other pathogens such as Babesia spp., Rickettsia spp., and Anaplasma phagocytophilum, are of increasing public health interest. In Poland, as in other European countries, Ixodes ricinus, the most prevalent tick species responsible for the majority of tick bites in humans, is the main vector of A. phagocytophilum. The aim of the study was to estimate the infection level of I. ricinus with A. phagocytophilum in selected districts, not previously surveyed for the presence of this agent. Sampling of questing ticks was performed in 12 forested sites, located in four districts (Legnica, Milicz, Lubań, and Oława) in SW Poland. Altogether, 792 ticks (151 females, 101 males, and 540 nymphs) representing I. ricinus were checked for the presence of A. phagocytophilum. The average infection level was 4.3%, with higher rate reported for adult ticks. The highest percentage of infected adults was observed in Milicz (17.4%) and the lowest in Oława (6.8%). The abundance of questing I. ricinus in all examined sites as well as the infection with A. phagocytophilum indicate for the first time the risk for HGA transmission in SW Poland.

Anaplasma phagocytophilum Manipulates Host Cell Apoptosis by Different Mechanisms to Establish Infection

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Pilar Alberdi

2016-07-01

Full Text Available Anaplasma phagocytophilum is an emerging zoonotic pathogen that causes human and animal granulocytic anaplasmosis and tick-borne fever of ruminants. This obligate intracellular bacterium evolved to use common strategies to establish infection in both vertebrate hosts and tick vectors. Herein, we discuss the different strategies used by the pathogen to modulate cell apoptosis and establish infection in host cells. In vertebrate neutrophils and human promyelocytic cells HL-60, both pro-apoptotic and anti-apoptotic factors have been reported. Tissue-specific differences in tick response to infection and differential regulation of apoptosis pathways have been observed in adult female midguts and salivary glands in response to infection with A. phagocytophilum. In tick midguts, pathogen inhibits apoptosis through the Janus kinase/signal transducers and activators of transcription (JAK/STAT pathway, while in salivary glands, the intrinsic apoptosis pathways is inhibited but tick cells respond with the activation of the extrinsic apoptosis pathway. In Ixodes scapularis ISE6 cells, bacterial infection down-regulates mitochondrial porin and manipulates protein processing in the endoplasmic reticulum and cell glucose metabolism to inhibit apoptosis and facilitate infection, whereas in IRE/CTVM20 tick cells, inhibition of apoptosis appears to be regulated by lower caspase levels. These results suggest that A. phagocytophilum uses different mechanisms to inhibit apoptosis for infection of both vertebrate and invertebrate hosts.

Merozoite surface protein-1 genetic diversity in Plasmodium malariae and Plasmodium brasilianum from Brazil.

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Guimarães, Lilian O; Wunderlich, Gerhard; Alves, João M P; Bueno, Marina G; Röhe, Fabio; Catão-Dias, José L; Neves, Amanda; Malafronte, Rosely S; Curado, Izilda; Domingues, Wilson; Kirchgatter, Karin

2015-11-16

The merozoite surface protein 1 (MSP1) gene encodes the major surface antigen of invasive forms of the Plasmodium erythrocytic stages and is considered a candidate vaccine antigen against malaria. Due to its polymorphisms, MSP1 is also useful for strain discrimination and consists of a good genetic marker. Sequence diversity in MSP1 has been analyzed in field isolates of three human parasites: P. falciparum, P. vivax, and P. ovale. However, the extent of variation in another human parasite, P. malariae, remains unknown. This parasite shows widespread, uneven distribution in tropical and subtropical regions throughout South America, Asia, and Africa. Interestingly, it is genetically indistinguishable from P. brasilianum, a parasite known to infect New World monkeys in Central and South America. Specific fragments (1 to 5) covering 60 % of the MSP1 gene (mainly the putatively polymorphic regions), were amplified by PCR in isolates of P. malariae and P. brasilianum from different geographic origin and hosts. Sequencing of the PCR-amplified products or cloned PCR fragments was performed and the sequences were used to construct a phylogenetic tree by the maximum likelihood method. Data were computed to give insights into the evolutionary and phylogenetic relationships of these parasites. Except for fragment 4, sequences from all other fragments consisted of unpublished sequences. The most polymorphic gene region was fragment 2, and in samples where this region lacks polymorphism, all other regions are also identical. The low variability of the P. malariae msp1 sequences of these isolates and the identification of the same haplotype in those collected many years apart at different locations is compatible with a low transmission rate. We also found greater diversity among P. brasilianum isolates compared with P. malariae ones. Lastly, the sequences were segregated according to their geographic origins and hosts, showing a strong genetic and geographic structure. Our data

Brotes de babesiosis y anaplasmosis bovina diagnosticados en el INTA Salta, Argentina: período 2006-2016

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ARAOZ V

2017-12-01

Full Text Available Se realizó un estudio retrospectivo de brotes de babesiosis y anaplasmosis (tristeza parasitaria bovina, TPB ocurridos entre 2006 y 2016 en el Noroeste argentino (NOA para evaluar algunos aspectos epidemiológicos y clínicos de estas enfermedades. De 4.398 muestras para diagnóstico, 238 (5,4% tuvieron sospechas de TPB, las cuales se confirmaron en 106 (44,3% casos por observación microscópica de los agentes causales. Cuarenta y siete (44,3% brotes se debieron a Anaplasma marginale, 40 (37,7% a Babesia bovis y 14 (13,2% a Babesia bigemina. En cinco (4,7% focos de babesiosis no se determinó la especie causal de Babesia. Los brotes se diagnosticaron en 62 establecimientos ganaderos de cuatro provincias (Salta, Jujuy, Santiago del Estero y Chaco. La mayoría ocurrieron en verano y otoño, con máxima incidencia en abril y nula en setiembre para ambas noxas. Las vacas adultas fueron la categoría de ganado más afectada. En 58% (61/106 de los brotes murió al menos un bovino, con un número promedio de cuatro muertes por brote. Se concluye que la TPB continúa incidiendo negativamente en la producción bovina del NOA. SUMMARY. Bovine babesiosis and anaplasmosis outbreaks diagnosed at INTA Salta, Argentina: 2006-2016. A retrospective study of babesiosis and anaplasmosis outbreaks (tick fever, TF was made, which occurred from 2006 to 2016 in Northwest Argentina (NWA, to evaluate clinical and epidemiological aspects of these diseases. From 4,398 specimens, 238 (5.4% were associated with suspicions of TF and 106 of these cases (44.3% were confirmed as TF outbreaks by microscopic observation of causal agents. Forty seven (44.3% were due to Anaplasma marginale, 40 (37.7% to Babesia bovis and 14 (13.2% to Babesia bigemina. In five (4.7% outbreaks of babesiosis, Babesia species were not determined. Outbreaks were diagnosed at 62 different cattle ranches in four provinces (Salta, Jujuy, Santiago del Estero and Chaco. The outbreaks occurred mostly during

Diversity and population structure of Plasmodium falciparum in Thailand based on the spatial and temporal haplotype patterns of the C-terminal 19-kDa domain of merozoite surface protein-1.

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Simpalipan, Phumin; Pattaradilokrat, Sittiporn; Siripoon, Napaporn; Seugorn, Aree; Kaewthamasorn, Morakot; Butcher, Robert D J; Harnyuttanakorn, Pongchai

2014-02-12

The 19-kDa C-terminal region of the merozoite surface protein-1 of the human malaria parasite Plasmodium falciparum (PfMSP-119) constitutes the major component on the surface of merozoites and is considered as one of the leading candidates for asexual blood stage vaccines. Because the protein exhibits a level of sequence variation that may compromise the effectiveness of a vaccine, the global sequence diversity of PfMSP-119 has been subjected to extensive research, especially in malaria endemic areas. In Thailand, PfMSP-119 sequences have been derived from a single parasite population in Tak province, located along the Thailand-Myanmar border, since 1995. However, the extent of sequence variation and the spatiotemporal patterns of the MSP-119 haplotypes along the Thai borders with Laos and Cambodia are unknown. Sixty-three isolates of P. falciparum from five geographically isolated populations along the Thai borders with Myanmar, Laos and Cambodia in three transmission seasons between 2002 and 2008 were collected and culture-adapted. The msp-1 gene block 17 was sequenced and analysed for the allelic diversity, frequency and distribution patterns of PfMSP-119 haplotypes in individual populations. The PfMSP-119 haplotype patterns were then compared between parasite populations to infer the population structure and genetic differentiation of the malaria parasite. Five conserved polymorphic positions, which accounted for five distinct haplotypes, of PfMSP-119 were identified. Differences in the prevalence of PfMSP-119 haplotypes were detected in different geographical regions, with the highest levels of genetic diversity being found in the Kanchanaburi and Ranong provinces along the Thailand-Myanmar border and Trat province located at the Thailand-Cambodia border. Despite this variability, the distribution patterns of individual PfMSP-119 haplotypes seemed to be very similar across the country and over the three malarial transmission seasons, suggesting that gene flow

CYP1A1 gene polymorphisms increase lung cancer risk in a high-incidence region of Spain: a case control study

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San Jose Carmen

2010-08-01

Full Text Available Abstract Background A rural region in south-west Spain has one of the highest lung cancer incidence rates of the country, as revealed by a previous epidemiological 10-year follow-up study. The present work was undertaken to ascertain the role of CYP1A1 gene polymorphisms and their interaction with tobacco smoking in the development of the disease in this location. Methods One-hundred-and-three cases of lung cancer and 265 controls participated in the study. The participants were screened for the presence of four CYP1A1 polymorphisms, namely MspI, Ile462Val, T3205C, and Thr461Asn. Lung cancer risk was estimated as odds ratios (OR and 95% confidence intervals (CI using unconditional logistic regression models adjusting for age, sex, and smoking. Results The distribution of the variant CYP1A1 alleles was different from that described for other Caucasian populations, with CYP1A1*2A showing an uncommonly high frequency (p CYP1A1*2B allele (carrying MspI and Ile462Val mutations was strongly associated with high lung cancer risk (OR = 4.59, CI:1.4-12.6, p p p = 0.04. Moreover, the Thr461Asn polymorphism was found to be associated with SCLC in a Caucasian population for the first time to our knowledge (OR = 8.33, CI: 1.3-15.2, p = 0.04. Conclusion The results suggest that CYP1A1 polymorphisms contribute to increase lung cancer susceptibility in an area with an uncommon high incidence rate.

Comparative strain analysis of Anaplasma phagocytophilum infection and clinical outcomes in a canine model of granulocytic anaplasmosis.

Science.gov (United States)

Scorpio, Diana G; Dumler, J Stephen; Barat, Nicole C; Cook, Judith A; Barat, Christopher E; Stillman, Brett A; DeBisceglie, Kristen C; Beall, Melissa J; Chandrashekar, Ramaswamy

2011-03-01

A pilot study was conducted to determine whether existing human or canine strains of Anaplasma phagocytophilum would reproduce clinical disease in experimentally inoculated dogs similar to dogs with naturally acquired granulocytic anaplasmosis. Six hounds were inoculated intravenously with one human and two canine strains of A. phagocytophilum that were propagated in vitro in HL-60 cells or in infected autologous neutrophils. Infected dogs were monitored for lethargy, anorexia, petechiae, lymphadenopathy, and fever. Dogs were assessed for complete blood count (CBC), serum chemistry, and serology (IFA and SNAP® 4Dx®); for A. phagocytophilum blood load by quantitative polymerase chain reaction; and for cytokine production. Prominent clinical signs were generalized lymphadenopathy and scleral injection; only one dog developed fever lasting 4 days. Notable laboratory alterations included sustained leukopenia and thrombocytopenia in all dogs. A. phagocytophilum morulae were noted in blood between days 10 and 11, although all dogs retained A. phagocytophilum DNA in blood through day 60. All dogs seroconverted by days 10-15 by IFA, and by days 17-30 by SNAP 4Dx; cytokine analyses revealed 10-fold increases in interleukin-2 and interleukin-18 in the neutrophil-propagated 98E4 strain-infected dog. All A. phagocytophilum strains produced infection, although canine 98E4 strain reproduced clinical signs, hematologic changes, and inflammatory cytokine elevations most consistent with granulocytic anaplasmosis when recognized clinically. Therefore, this strain should be considered for use in future studies of A. phagocytophilum canine infection models.

Genetic polymorphisms in CYP1A1, GSTM1, GSTP1 and GSTT1 metabolic genes and risk of lung cancer in Asturias

International Nuclear Information System (INIS)

López-Cima, M Felicitas; Álvarez-Avellón, Sara M; Pascual, Teresa; Fernández-Somoano, Ana; Tardón, Adonina

2012-01-01

Metabolic genes have been associated with the function of metabolizing and detoxifying environmental carcinogens. Polymorphisms present in these genes could lead to changes in their metabolizing and detoxifying ability and thus may contribute to individual susceptibility to different types of cancer. We investigated if the individual and/or combined modifying effects of the CYP1A1 MspI T6235C, GSTM1 present/null, GSTT1 present/null and GSTP1 Ile105Val polymorphisms are related to the risk of developing lung cancer in relation to tobacco consumption and occupation in Asturias, Northern Spain. A hospital-based case–control study (CAPUA Study) was designed including 789 lung cancer patients and 789 control subjects matched in ethnicity, age, sex, and hospital. Genotypes were determined by PCR or PCR-RFLP. Individual and combination effects were analysed using an unconditional logistic regression adjusting for age, pack-years, family history of any cancer and occupation. No statistically significant main effects were observed for the carcinogen metabolism genes in relation to lung cancer risk. In addition, the analysis did not reveal any significant gene-gene, gene-tobacco smoking or gene-occupational exposure interactions relative to lung cancer susceptibility. Lastly, no significant gene-gene combination effects were observed. These results suggest that genetic polymorphisms in the CYP1A1, GSTM1, GSTT1 and GSTP1 metabolic genes were not significantly associated with lung cancer risk in the current study. The results of the analysis of gene-gene interactions of CYP1A1 MspI T6235C, GSTM1 present/null, GSTT1 present/null and GSTP1 Ile105Val polymorphisms in lung cancer risk indicate that these genes do not interact in lung cancer development

The development of an enzyme-linked immunosorbent assay for Trypanosoma vivax antibodies and its use in epidemiological surveys

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Claudio R Madruga

2006-11-01

Full Text Available There are data indicating that the distribution of Trypanosoma vivax in the Brazilian territory is expanding with potential to reach other areas, where the vectors are present. The detection of anti-trypanosomal antibodies in serum provides important information of the trypanosomal status in cattle herds. For this reason, an enzyme-linked immunosorbent assay (Tv-ELISA-Ab with crude antigen from one Brazilian isolate of T. vivax was developed and evaluated. The sensitivity and specificity were respectively 97.6 and 96.9%. In the evaluation of cross-reactions, three calves inoculated with T. evansi trypimastigotes blood forms showed optical densities (OD under the cut-off during the whole experimental period, except one at 45 days post-inoculation. With relation to Babesia bovis, B. bigemina, and Anaplasma marginale, which are endemic hemoparasites in the studied area, the cross-reactions were shown to be 5.7, 5.3, and 1.1%, respectively. The first serological survey of Pantanal and state of Pará showed that T. vivax is widespread, although regions within both areas had significantly different prevalences. Therefore, this Tv-ELISA-Ab may be a more appropriate test for epidemiological studies in developing countries because the diagnostic laboratories in most countries may be able to perform an ELISA, which is not true for polymerase chain reaction.

First molecular evidence of Anaplasma ovis and Rickettsia spp. in keds (Diptera: Hippoboscidae) of sheep and wild ruminants.

Science.gov (United States)

Hornok, Sándor; de la Fuente, José; Biró, Nóra; Fernández de Mera, Isabel G; Meli, Marina L; Elek, Vilmos; Gönczi, Eniko; Meili, Theres; Tánczos, Balázs; Farkas, Róbert; Lutz, Hans; Hofmann-Lehmann, Regina

2011-10-01

To evaluate the presence of rickettsial agents in hippoboscid flies with molecular methods, 81 sheep keds (Melophagus ovinus) were collected from 23 sheep, 144 deer keds (Lipoptena cervi) were caught in the environment, and a further 463 and 59 individuals of the latter species were obtained from fresh carcasses of 29 red deer and 17 roe deer, respectively. DNA was extracted individually or in pools. Anaplasma ovis was demonstrated in all examined sheep keds, and from one pool of free-living deer keds. Rickettsia helvetica or other, unidentified rickettsiae were also present in one pool of sheep keds, and in four pools of deer keds from both red deer and roe deer. This is the first account of polymerase chain reaction positivity of hippoboscid flies for A. ovis and rickettsiae. These results raise the possibility that-apart from cattle and roe deer as already reported-sheep and red deer might also play a reservoir role in the epidemiology of rickettsioses.

Bmcystatin, a cysteine proteinase inhibitor characterized from the tick Boophilus microplus

International Nuclear Information System (INIS)

Lima, Cassia A.; Sasaki, Sergio D.; Tanaka, Aparecida S.

2006-01-01

The bovine tick Rhipicephalus (Boophilus) microplus is a blood-sucking animal, which is responsible for Babesia spp and Anaplasma marginale transmission for cattle. From a B. microplus fat body cDNA library, 465 selected clones were sequenced randomly and resulted in 60 Contigs. An open reading frame (ORF) contains 98 amino acids named Bmcystatin, due to 70% amino acid identity to a classical type 1 cystatin from Ixodes scapularis tick (GenBank Accession No. DQ066227). The Bmcystatin amino acid sequence analysis showed two cysteine residues, theoretical pI of 5.92 and M r of 11kDa. Bmcystatin gene was cloned in pET 26b vector and the protein expressed using bacteria Escherichia coli BL21 SI. Recombinant Bmcystatin (rBmcystatin) purified by affinity chromatography on Ni-NTA-agarose column and ionic exchange chromatography on HiTrap Q column presented molecular mass of 11kDa, by SDS-PAGE and the N-terminal amino acid sequenced revealed unprocessed N-terminal containing part of pelB signal sequence. Purified rBmcystatin showed to be a C1 cysteine peptidase inhibitor with K i value of 0.1 and 0.6nM for human cathepsin L and VTDCE (vitellin degrading cysteine endopeptidase), respectively. The rBmcystatin expression analyzed by semi-quantitative RT-PCR confirmed the amplification of a specific DNA sequence (294bp) in the fat body and ovary cDNA preparation. On the other hand, a protein band was detected in the fat body, ovary, and the salivary gland extracts using anti-Bmcystatin antibody by Western blot. The present results suggest a possible role of Bmcystatin in the ovary, even though the gene was cloned from the fat body, which could be another site of this protein synthesis

Physical workload, leisure-time physical activity, obesity and smoking as predictors of multisite musculoskeletal pain. A 2-year prospective study of kitchen workers.

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Haukka, Eija; Ojajärvi, Anneli; Takala, Esa-Pekka; Viikari-Juntura, Eira; Leino-Arjas, Päivi

2012-07-01

The aim of this prospective study was to examine the role of physical workload, leisure-time physical activity, obesity and smoking in predicting the occurrence and course of multisite musculoskeletal pain (MSP). Data on physical and psychosocial workload, lifestyle factors and MSP were based on questionnaire surveys of 385 Finnish female kitchen workers. MSP (defined as pain at three or more of seven sites) during the past 3 months was measured repeatedly at 3-month intervals over 2 years. Four different patterns (trajectories) in the course of MSP were identified. The authors analysed whether the determinants at baseline predicted the occurrence of MSP (1) at the 2-year follow-up and (2) over the total of nine measurements during the 2 years by exploiting the MSP trajectories. Logistic regression was used. High physical workload at baseline was an independent predictor of MSP at the 2-year follow-up (OR 3.8, 95% CI 1.7 to 8.5) in a model allowing for age, psychosocial factors at work and lifestyle. High physical workload (OR 2.0, 95% CI 1.0 to 4.0) and moderate (OR 2.4, 95% CI 1.2 to 4.9) or low (OR 2.3, 95% CI 1.1 to 4.7) physical activity predicted persistent MSP. Obesity (OR 2.8, 95% CI 1.0 to 7.8) predicted an increased, and not being obese (OR 3.7, 95% CI 1.1 to 12.7) a decreased, prevalence of MSP in models similarly including all covariates. Smoking had no effect. The results emphasise the importance of high physical workload, low to moderate physical activity and obesity as potential modifiable risk factors for the occurrence and course of MSP over time.

Genetic variation in genes for the xenobiotic-metabolizing enzymes CYP1A1, EPHX1, GSTM1, GSTT1 and GSTP1 and susceptibility to colorectal cancer in Lynch syndrome

Science.gov (United States)

Pande, Mala; Amos, Christopher I.; Osterwisch, Daniel R.; Chen, Jinyun; Lynch, Patrick M.; Broaddus, Russell; Frazier, Marsha L.

2011-01-01

Individuals with Lynch syndrome are predisposed to cancer due to an inherited DNA mismatch repair gene mutation. However, there is significant variability observed in disease expression, likely due to the influence of other environmental, lifestyle, or genetic factors. Polymorphisms in genes encoding xenobiotic-metabolizing enzymes may modify cancer risk by influencing the metabolism and clearance of potential carcinogens from the body. In this retrospective analysis, we examined key candidate gene polymorphisms in CYP1A1, EPHX1, GSTT1, GSTM1, and GSTP1 as modifiers of age at onset of colorectal cancer among 257 individuals with Lynch syndrome. We found that subjects heterozygous for CYP1A1 I462V (c.1384A>G) developed colorectal cancer 4 years earlier than those with the homozygous wild-type genotype (median ages 39 and 43 years, respectively; log-rank test P = 0.018). Furthermore, being heterozygous for the CYP1A1 polymorphisms, I462V and Msp1 (g.6235T>C), was associated with an increased risk for developing colorectal cancer [adjusted hazard ratio for AG relative to AA = 1.78, 95% CI = 1.16–2.74, P = 0.008; and hazard ratio for TC relative to TT = 1.53, 95% CI = 1.06–2.22, P = 0.02]. Since homozygous variants for both CYP1A1 polymorphisms were rare, risk estimates were imprecise. None of the other gene polymorphisms examined were associated with an earlier onset age for colorectal cancer. Our results suggest that the I462V and Msp1 polymorphisms in CYP1A1 may be an additional susceptibility factor for disease expression in Lynch syndrome since they modify the age of colorectal cancer onset by up to 4 years. PMID:18768509

Molecular detection of Anaplasma platys, Ehrlichia canis, Hepatozoon canis and Rickettsia monacensis in dogs from Maio Island of Cape Verde archipelago.

Science.gov (United States)

Lauzi, Stefania; Maia, João P; Epis, Sara; Marcos, Ricardo; Pereira, Cristina; Luzzago, Camilla; Santos, Marta; Puente-Payo, Pablo; Giordano, Alessia; Pajoro, Massimo; Sironi, Giuseppe; Faustino, Augusto

2016-07-01

Tick-borne diseases are emerging worldwide and have an important zoonotic relevance. Dogs play an important role in the epidemiology of several zoonotic tick-borne pathogens acting as sentinels and/or reservoirs. This study focused on the molecular identification of tick-borne pathogens in blood samples of 153 autochthonous asymptomatic dogs in Maio Island, Cape Verde archipelago. Eighty-four (54.9%) dogs were positive for one or more pathogens. Fifty-five (35.9%) dogs were infected with Hepatozoon canis, 53 (34.6%) with Anaplasma platys, five (3.3%) with Ehrlichia canis and Rickettsia monacensis, an emerging human pathogen, was also identified in a single dog (0.7%). The former three pathogens cause important canine tick-borne diseases that are transmitted or potentially transmitted by Rhipicephalus sanguineus s.l., the only hard tick identified in Cape Verde. Furthermore, Wolbachia spp. was amplified from the blood of one dog. None of the dogs were positive for Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, Midichloria mitochondrii, Bartonella spp., Babesia spp. or Theileria spp. Fifty-four (35.3%) animals showed single infections and 30 (19.6%) co-infections, with A. platys and H. canis co-infection being the most frequent (28 dogs, 18.3%). The frequency of E. canis infection was statistically different among age groups (P=0.017), being higher among dogs older than 4 years compared to younger dogs. Infection by A. platys was also statistically different among age groups (P=0.031), being higher in dogs younger than 2 years compared to older dogs. The statistical analyses showed no significant association of PCR positivity with gender or location. The frequency of tick-borne pathogens detected in dogs in Maio Island, including R. monacensis, highlights the need to improve diagnosis and control in order to prevent the risk of transmission of these pathogens among dogs and humans living in or travelling to this touristic island. Copyright © 2016

Ribosomal Protein S6 Kinase (RSK-2 as a central effector molecule in RON receptor tyrosine kinase mediated epithelial to mesenchymal transition induced by macrophage-stimulating protein

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Zhang Rui-Wen

2011-05-01

Full Text Available Abstract Background Epithelial to mesenchymal transition (EMT occurs during cancer cell invasion and malignant metastasis. Features of EMT include spindle-like cell morphology, loss of epithelial cellular markers and gain of mesenchymal phenotype. Activation of the RON receptor tyrosine kinase by macrophage-stimulating protein (MSP has been implicated in cellular EMT program; however, the major signaling determinant(s responsible for MSP-induced EMT is unknown. Results The study presented here demonstrates that RSK2, a downstream signaling protein of the Ras-Erk1/2 pathway, is the principal molecule that links MSP-activated RON signaling to complete EMT. Using MDCK cells expressing RON as a model, a spindle-shape based screen was conducted, which identifies RSK2 among various intracellular proteins as a potential signaling molecule responsible for MSP-induced EMT. MSP stimulation dissociated RSK2 with Erk1/2 and promoted RSK2 nuclear translocation. MSP strongly induced RSK2 phosphorylation in a dose-dependent manner. These effects relied on RON and Erk1/2 phosphorylation, which is significantly potentiated by transforming growth factor (TGF-β1, an EMT-inducing cytokine. Specific RSK inhibitor SL0101 completely prevented MSP-induced RSK phosphorylation, which results in inhibition of MSP-induced spindle-like morphology and suppression of cell migration associated with EMT. In HT-29 cancer cells that barely express RSK2, forced RSK2 expression results in EMT-like phenotype upon MSP stimulation. Moreover, specific siRNA-mediated silencing of RSK2 but not RSK1 in L3.6pl pancreatic cancer cells significantly inhibited MSP-induced EMT-like phenotype and cell migration. Conclusions MSP-induced RSK2 activation is a critical determinant linking RON signaling to cellular EMT program. Inhibition of RSK2 activity may provide a therapeutic opportunity for blocking RON-mediated cancer cell migration and subsequent invasion.

Resolving the paradox for protein aggregation diseases: NMR structure and dynamics of the membrane-embedded P56S-MSP causing ALS imply a common mechanism for aggregation-prone proteins to attack membranes [v2; ref status: indexed, http://f1000r.es/3zl

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Haina Qin

2014-07-01

Full Text Available Paradoxically, aggregation of specific proteins is characteristic of many human diseases and aging, yet aggregates have increasingly been found to be unnecessary for initiating pathogenesis. Here we determined the NMR topology and dynamics of a helical mutant in a membrane environment transformed from the 125-residue cytosolic all-β MSP domain of vesicle-associated membrane protein-associated protein B (VAPB by the ALS-causing P56S mutation. Despite its low hydrophobicity, the P56S major sperm protein (MSP domain becomes largely embedded in the membrane environment with high backbone rigidity. Furthermore it is composed of five helices with amphiphilicity comparable to those of the partly-soluble membrane toxin mellitin and α-synuclein causing Parkinson's disease. Consequently, the mechanism underlying this chameleon transformation becomes clear: by disrupting the specific tertiary interaction network stabilizing the native all-β MSP fold to release previously-locked amphiphilic segments, the P56S mutation acts to convert the classic MSP fold into a membrane-active protein that is fundamentally indistinguishable from mellitin and α-synuclein which are disordered in aqueous solution but spontaneously partition into membrane interfaces driven by hydrogen-bond energetics gained from forming α-helix in the membrane environments. As segments with high amphiphilicity exist in all proteins, our study successfully resolves the paradox by deciphering that the proteins with a higher tendency to aggregate have a stronger potential to partition into membranes through the same mechanism as α-synuclein to initially attack membranes to trigger pathogenesis without needing aggregates. This might represent the common first step for various kinds of aggregated proteins to trigger familiar, sporadic and aging diseases. Therefore the homeostasis of aggregated proteins in vivo is the central factor responsible for a variety of human diseases including aging

Non-additive effects of RBP4, ESR1 and IGF2 polymorphisms on litter size at different parities in a Chinese-European porcine line

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Alves Estefânia

2010-06-01

Full Text Available Abstract Background The aim of this work was to study the effects on litter size of variants of the porcine genes RBP4, ESR1 and IGF2, currently used in genetic tests for different purposes. Moreover, we investigated a possible effect of the interaction between RBP4-MspI and ESR1-PvuII polymorphisms. The IGF2-intron3-G3072A polymorphism is actually used to select lean growth, but other possible effects of this polymorphism on reproductive traits need to be evaluated. Methods Detection of polymorphisms in the genomic and cDNA sequences of RBP4 gene was carried out. RBP4-MspI and IGF2-intron3-G3072A were genotyped in a hyperprolific Chinese-European line (Tai-Zumu and three new RBP4 polymorphisms were genotyped in different pig breeds. A bivariate animal model was implemented in association analyses considering the number of piglets born alive at early (NBA12 and later parities (NBA3+ as different traits. A joint analysis of RBP4-MspI and ESR1-PvuII was performed to test their possible interaction. In the IGF2 analysis, paternal or maternal imprinting effects were also considered. Results Four different RBP4 haplotypes were detected (TGAC, GGAG, GAAG and GATG in different pig breeds and wild boars. A significant interaction effect between RBP4-MspI and ESR1-PvuII polymorphisms of 0.61 ± 0.29 piglets was detected on NBA3+. The IGF2 analysis revealed a significant increase on NBA3+ of 0.74 ± 0.37 piglets for the paternally inherited allele A. Conclusions All the analyzed pig and wild boar populations shared one of the four detected RBP4 haplotypes. This suggests an ancestral origin of the quoted haplotype. The joint use of RBP4-MspI and ESR1-PvuII polymorphisms could be implemented to select for higher prolificacy in the Tai-Zumu line. In this population, the paternal allele IGF2-intron3-3072A increased litter size from the third parity. The non-additive effects on litter size reported here should be tested before implementation in other pig

Análise comparativa entre as metodologias de PCR metilação-específica (MSP, Southern blot (SB e FISH utilizadas no diagnóstico genético molecular de pacientes com suspeita clínica das síndromes de Prader-Willi ou Angelman

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Bruna Oliveira

2016-08-01

Full Text Available Introdução: Prader-Willi (SPW e Angelman (SA são síndromes clinicamente distintas, causadas pela perda de expressão de genes na região cromossômica 15q11.2-q13, de origem paterna ou materna, respectivamente. Ambas compartilham os mesmos métodos diagnósticos. Nossos objetivos foram: a analisar por PCR metilação-específica (MSP pacientes com suspeita clínica de SPW/SA; b comparar resultados de diferentes metodologias de diagnóstico molecular; c aplicar a técnica MSP na rotina assistencial de pacientes encaminhados ao Serviço de Genética Médica/Hospital de Clínicas de Porto Alegre (SGM/HCPA. Métodos: Foram analisados 123 pacientes com suspeita clínica de SPW (n = 71 ou SA (n = 52 por MSP. Desses, 79 possuíam análise prévia por hibridação in situ fluorescente (FISH e/ou Southern blot (SB. Resultados: Foram detectados 21 casos positivos – 15 de SPW (12,19% e 6 de SA (4,88%. Nove pacientes tiveram etiologia molecular determinada, sendo sete com diagnóstico de SPW (quatro dissomias uniparentais – UPD15 materna – e três deleções na região 15q11-13 e dois com diagnóstico de SA (um com UPD15 paterna e um com deleção na região 15q11-13. Foram observados resultados equivalentes entre MSP e SB e resultados discrepantes entre MSP e FISH (n = 4. Foram padronizados dois protocolos de MSP para confirmação dos resultados e controle interno de qualidade. Conclusão: O perfil de detecção de cada técnica varia de acordo com o mecanismo etiológico presente. A análise por MSP detecta alterações no padrão de metilação geradas por deleção, UPD e defeitos de imprinting, sem identificar o mecanismo etiológico responsável. Contudo, mostrou ser eficiente para confirmação do diagnóstico clínico e screening dos pacientes com suspeita clínica sugestiva de SPW e SA. Diante de resultados positivos, é importante a identificação do mecanismo molecular subjacente para correlação genótipo-fenótipo e determina

Study of mixed programming gamma spectrum acquisition method based on MSP430F4618

International Nuclear Information System (INIS)

Li Yuezhong; Tang Bin; Zhang Zhongliang; Xie Xiaolin

2012-01-01

In order to reduce the hand-held gamma spectrometer measurements dead time and to complete the low-voltage and low-power design, the spectrum signal acquisition circuit is constituted by the ultra-low power microcontroller MSP430F4618 and its external signal conditioning circuit, anti-coincidence circuit interface and its on-chip sample hold and A/D converter. C language programming and assembly one have been used together. The sample hold and A/D conversion and spectrum acquisition programming is accomplished by assembly language, and the system monitoring and task scheduler designing is accomplished by C language programming. The handhold gamma spectrometer power supply, which just uses two No.5 rechargeable batteries, is designed by high-efficiency DC-DC circuit. The prototype gamma spectrometer is developed by the method, and its testing shows that the implementation of spectrum acquisition time is shorten by twice to 3 times, that is, the dead measurement can be reduced; and the machine operating current does not exceed 150 mA. By using two 2400 mAh No.5 rechargeable battery, the machine can work continuously more than 10 hours, and it can meet the application requirements. (authors)

Levels of antibody to conserved parts of Plasmodium falciparum merozoite surface protein 1 in Ghanaian children are not associated with protection from clinical malaria

DEFF Research Database (Denmark)

Dodoo, D; Theander, T G; Kurtzhals, J A

1999-01-01

malaria season in April and after the season in November. Using enzyme-linked immunosorbent assay, we measured antibody responses to recombinant gluthathione S-transferase-PfMSP119 fusion proteins corresponding to the Wellcome and MAD20 allelic variants in these samples. Prevalence of antibodies......The 19-kDa conserved C-terminal part of the Plasmodium falciparum merozoite surface protein 1 (PfMSP119) is a malaria vaccine candidate antigen, and human antibody responses to PfMSP119 have been associated with protection against clinical malaria. In this longitudinal study carried out in an area...

(igf1/igf1r) with milk production tr

African Journals Online (AJOL)

Gosia

2016-06-15

Jun 15, 2016 ... fragment length polymorphism (PCR-RFLP) (TaiI and MspI restriction enzymes) and amplification-created restriction site (ACRS) (SnaBI ... is the first association study based on polymorphisms of the primary genes encoding the IGF-1 system in a small herd of .... However, protein content was highest in milk ...

Real-time colorimetric detection of DNA methylation of the PAX1 gene in cervical scrapings for cervical cancer screening with thiol-labeled PCR primers and gold nanoparticles

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Huang J

2016-10-01

Full Text Available Jin Huang,1,2 Yu-Ligh Liou,1,2 Ya-Nan Kang,3 Zhi-Rong Tan,1,2 Ming-Jing Peng,1,2 Hong-Hao Zhou1,2 1Department of Clinical Pharmacology, Xiangya Hospital, Central South University, 2Institute of Clinical Pharmacology, Hunan Key Laboratory of Pharmacogenetics, Central South University, 3Department of Obstetrics and Gynecology, Xiangya Hospital, Central South University, Changsha, Hunan, People’s Republic of China Background: DNA methylation can induce carcinogenesis by silencing key tumor suppressor genes. Analysis of aberrant methylation of tumor suppressor genes can be used as a prognostic and predictive biomarker for cancer. In this study, we propose a colorimetric method for the detection of DNA methylation of the paired box gene 1 (PAX1 gene in cervical scrapings obtained from 42 patients who underwent cervical colposcopic biopsy. Methods: A thiolated methylation-specific polymerase chain reaction (MSP primer was used to generate MSP products labeled with the thiol group at one end. After bisulfite conversion and MSP amplification, the unmodified gold nanoparticles (AuNPs were placed in a reaction tube and NaCl was added to induce aggregation of bare AuNPs without generating polymerase chain reaction products. After salt addition, the color of AuNPs remained red in the methylated PAX1 gene samples because of binding to the MSP-amplified products. By contrast, the color of the AuNP colloid solution changed from red to blue in the non-methylated PAX1 gene samples because of aggregation of AuNPs in the absence of the MSP-amplified products. Furthermore, PAX1 methylation was quantitatively detected in cervical scrapings of patients with varied pathological degrees of cervical cancer. Conventional quantitative MSP (qMSP was also performed for comparison. Results: The two methods showed a significant correlation of the methylation frequency of the PAX1 gene in cervical scrapings with severity of cervical cancer (n=42, P<0.05. The results of the

Molecular epidemiology of Anaplasma platys, Ehrlichia canis and Babesia vogeli in stray dogs in Paraná, Brazil

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Claudia M. Ribeiro

Full Text Available ABSTRACT: Hemoparasitic infections are tick-borne diseases, which affect animals and humans. Considering the importance of canine hemoparasitic infections in veterinary clinics, this study aimed to determine the occurrence of Anaplasma platys, Ehrlichia canis and Babesia vogeli in blood samples from 182 dogs not domiciled in the city of Pato Branco, southwestern region of Paraná State, Brazil, using polymerase chain reaction (PCR. The prevalence of A. platys and B. vogeli was 32.9% and 10.9% respectively, and A. platys infection prevailed (p<0.001. The number of dogs positive for A. platys was larger in Winter (p<0.05. All blood samples were negative for E. canis. In the dogs, infestation by Amblyomma cajennense predominated over that by Rhipicephalus sanguineus (p<0.001; but there was no significant association between PCR and the variables presence of ticks, sex and age. Dogs infected by A. platys and B. vogeli showed thrombocytopenia, lymphopenia and leukocytosis; but there was no correlation between such hematological changes and infection by hemoparasites. This appears to be the first molecular study that demonstrates the existence of A. platys and B. vogeli in dogs from the southwestern region of Paraná.

Parasites of domestic and wild animals in South Africa. L. Ixodid ticks infesting horses and donkeys.

Science.gov (United States)

Horak, Ivan G; Heyne, Heloise; Halajian, Ali; Booysen, Shalaine; Smit, Willem J

2017-02-28

The aim of the study was to determine the species spectrum of ixodid ticks that infest horses and donkeys in South Africa and to identify those species that act as vectors of disease to domestic livestock. Ticks were collected opportunistically from 391 horses countrywide by their owners or grooms, or by veterinary students and staff at the Faculty of Veterinary Science, University of Pretoria. Ticks were also collected from 76 donkeys in Limpopo Province, 2 in Gauteng Province and 1 in North West province. All the ticks were identified by means of a stereoscopic microscope. Horses were infested with 17 tick species, 72.1% with Rhipicephalus evertsi evertsi, 19.4% with Amblyomma hebraeum and 15.6% with Rhipicephalus decoloratus. Rhipicephalus evertsi evertsi was recovered from horses in all nine provinces of South Africa and R. decoloratus in eight provinces. Donkeys were infested with eight tick species, and 81.6% were infested with R. evertsi evertsi, 23.7% with A. hebraeum and 10.5% with R. decoloratus. Several tick species collected from the horses and donkeys are the vectors of economically important diseases of livestock. Rhipicephalus evertsi evertsi is the vector of Theileria equi, the causative organism of equine piroplasmosis. It also transmits Anaplasma marginale, the causative organism of anaplasmosis in cattle. Amblyomma hebraeum is the vector of Ehrlichia ruminantium, the causative organism of heartwater in cattle, sheep and goats, whereas R. decoloratus transmits Babesia bigemina, the causative organism of babesiosis in cattle.

The Suitability of P. falciparum Merozoite Surface Proteins 1 and 2 as Genetic Markers for In Vivo Drug Trials in Yemen

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Al-abd, Nazeh M.; Mahdy, Mohammed A. K.; Al-Mekhlafi, Abdulsalam M. Q.; Snounou, Georges; Abdul-Majid, Nazia B.; Al-Mekhlafi, Hesham M.; Fong, Mun Y.

2013-01-01

Background The accuracy of the conclusions from in vivo efficacy anti-malarial drug trials depends on distinguishing between recrudescences and re-infections which is accomplished by genotyping genes coding P. falciparum merozoite surface 1 (MSP1) and MSP2. However, the reliability of the PCR analysis depends on the genetic markers’ allelic diversity and variant frequency. In this study the genetic diversity of the genes coding for MSP1 and MSP2 was obtained for P. falciparum parasites circulating in Yemen. Methods Blood samples were collected from 511 patients with fever and screened for malaria parasites using Giemsa-stained blood films. A total 74 samples were infected with P. falciparum, and the genetic diversity was assessed by nested PCR targeting Pfmsp1 (Block2) and Pfmsp2 (block 3). Results Overall, 58%, 28% and 54% of the isolates harboured parasites of the Pfmsp1 K1, MAD20 and RO33 allelic families, and 55% and 89% harboured those of the Pfmsp2 FC27 and 3D7 allelic families, respectively. For both genetic makers, the multiplicity of the infection (MOI) was significantly higher in the isolates from the foothills/coastland areas as compared to those from the highland (PYemen Pfmsp1 should not be used for PCR correction of in vivo clinical trials outcomes, and that caution should be exercised when employing Pfmsp2. PMID:23861823

Detection of methylated CDO1 in plasma of colorectal cancer; a PCR study.

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Keishi Yamashita

Full Text Available BACKGROUND: Cysteine biology is important for the chemosensitivity of cancer cells. Our research has focused on the epigenetic silencing of cysteine dioxygenase type 1 (CDO1 in colorectal cancer (CRC. In this study, we describe detection of CDO1 methylation in the plasma of CRC patients using methylation specific PCR (Q-MSP and extensive analysis of the PCR reaction. METHODS: DNA was extracted from plasma, and analysed for methylation of the CDO1 gene using Q-MSP. The detection rate of CDO1 gene methylation was calculated and compared with that of diluted DNA extracted from "positive control" DLD1 cells. CDO1 gene methylation in the plasma of 40 CRC patients that were clinicopathologically analysed was then determined. RESULTS: (1 The cloned sequence analysis detected 93.3% methylation of the promoter CpG islands of the CDO1 gene of positive control DLD1 cells and 4.7% methylation of the negative control HepG2 CDO1 gene. (2 DLD1 CDO1 DNA could not be detected in this assay if the extracted DNA was diluted ∼1000 fold. The more DNA that was used for the PCR reaction, the more effectively it was amplified in Q-MSP. (3 By increasing the amount of DNA used, methylated CDO1 could be clearly detected in the plasma of 8 (20% of the CRC patients. However, the percentage of CRC patients detected by methylated CDO1 in plasma was lower than that detected by CEA (35.9% or CA19-9 (23.1% in preoperative serum. Combination of CEA/CA19-9 plus plasma methylated CDO1 could increase the rate of detection of curable CRC patients (39.3% as compared to CEA/CA19-9 (25%. CONCLUSION: We have described detection of CDO1 methylation in the plasma of CRC patients. Although CDO1 methylation was not detected as frequently as conventional tumor markers, analysis of plasma CDO1 methylation in combination with CEA/CA19-9 levels increases the detection rate of curable CRC patients.

NCBI nr-aa BLAST: CBRC-GGOR-01-1354 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-GGOR-01-1354 ref|YP_003328481.1| hypothetical protein ACIS_00561 [Anaplasma centrale str. Israel...] gb|ACZ49167.1| hypothetical protein ACIS_00561 [Anaplasma centrale str. Israel] YP_003328481.1 0.002 28% ...

NCBI nr-aa BLAST: CBRC-GGOR-01-1354 [SEVENS

Lifescience Database Archive (English)

Full Text Available CBRC-GGOR-01-1354 ref|YP_003328565.1| hypothetical protein ACIS_00677 [Anaplasma centrale str. Israel...] gb|ACZ49251.1| hypothetical protein ACIS_00677 [Anaplasma centrale str. Israel] YP_003328565.1 0.002 28% ...

Molecular characterization of Plasmodium falciparum in Arunachal Pradesh from Northeast India based on merozoite surface protein 1 & glutamate-rich protein.

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Sarmah, Nilanju Pran; Sarma, Kishore; Bhattacharyya, Dibya Ranjan; Sultan, Ali; Bansal, Devendra; Singh, Neeru; Bharti, Praveen K; Kaur, Hargobinder; Sehgal, Rakesh; Mohapatra, Pradyumna Kishore; Mahanta, Jagadish

2017-09-01

Northeast (NE) India is one of the high endemic regions for malaria with a preponderance of Plasmodium falciparum, resulting in high morbidity and mortality. The P. falciparum parasite of this region showed high polymorphism in drug-resistant molecular biomarkers. However, there is a paucity of information related to merozoite surface protein 1 (msp-1) and glutamate-rich protein (glurp) which have been extensively studied in various parts of the world. The present study was, therefore, aimed at investigating the genetic diversity of P. falciparum based on msp-1 and glurp in Arunachal Pradesh, a State in NE India. Two hundred and forty nine patients with fever were screened for malaria, of whom 75 were positive for P. falciparum. Blood samples were collected from each microscopically confirmed patient. The DNA was extracted; nested polymerase chain reaction and sequencing were performed to study the genetic diversity of msp-1 (block 2) and glurp. The block 2 of msp-1 gene was found to be highly polymorphic, and overall allelic distribution showed that RO33 was the dominant allele (63%), followed by MAD20 (29%) and K1 (8%) alleles. However, an extensive diversity (9 alleles and 4 genotypes) and 6-10 repeat regions exclusively of R2 type were observed in glurp. The P. falciparum population of NE India was diverse which might be responsible for higher plasticity leading to the survival of the parasite and in turn to the higher endemicity of falciparum malaria of this region.

SEROLOGICAL EVIDENCE OF EXPOSURE TO Anaplasma phagocytophilum IN HORSES FROM THE RIO DE JANEIRO STATE MOUNTED POLICE BRED IN THE URBAN ZONE

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Marcia Farias Rolim

2015-07-01

Full Text Available Anaplasma phagocytophilum is an emergent hemoparasite in regions where ticks are regularly found. In order to investigate the rate of anti-A. phagocytophilum antibody in horses of the Mounted Police of Rio de Janeiro state and therefore the presence of this agent, 41 horses from the Cavalry Squadron (CS and 50 from the Regiment of Mounted Police (RMP were selected. For the serologic diagnosis the Indirect Fluorescent Antibody test was performed. In the present work, among 91 equine sera samples anti-A. phagocytophilum antibodies, with titer ≥ 1:80, were detected in 11 animals (12%, being five (5% from males and six (7% females, with no statistical difference. The adult animals with ages varying between five and 14 years presented the highest rate of positive reaction, although antibodies were detected in animals of all ages. We did not observe statistic differences in relation to the presence of anti-A. phagocytophilum antibodies among the animals bred in the CS and RMP. The presence of anti-A.phagocytophilim antibodies in horses of the Mounted Police with no clinical signs is indicative that the parasite is present in the enzootic form among the horses of the urban area. The circulation of the parasite among the animals is not dependent of the presence of tick infestations.

Good work ability despite multisite musculoskeletal pain? A study among occupationally active Finns.

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Pensola, Tiina; Haukka, Eija; Kaila-Kangas, Leena; Neupane, Subas; Leino-Arjas, Päivi

2016-05-01

Although multisite pain (MSP) often threatens work ability (WA), some of those with MSP retain good WA. Our aim was to identify factors associated with good WA among subjects with MSP. A nationally representative sample (the Health 2000-Study, response rate 87%) comprising 3884 occupationally active Finns aged 30-64 years. Data on WA, musculoskeletal pain, physical and psychosocial working conditions, chronic diseases, lifestyle and domestic situation were gathered by questionnaire, interview and clinical examination. Good current WA compared with the lifetime best was defined as ⩾9 on a 0-10 scale. Musculoskeletal pain in 18 body locations was combined into four sites, and thereafter pain in two or more sites was defined as MSP (N=1351). Poisson regression analysis was used to obtain prevalence rate ratios (PRR). Good WA was reported by 48% of the women and 37% of the men with MSP. In a multivariable model good WA was associated with younger age, female gender, physically non-strenuous work (PRR 1.3, 95% CI 1.1-1.5), low job strain (1.2, 1.0-1.4), high supervisor support (1.2, 1.0-1.4), and not having musculoskeletal diseases (1.3, 1.1-1.5), mental disorders (1.4, 1.1-1.9), daytime tiredness (1.4, 1.2-1.7) or economic troubles (1.5, 1.1-1.9). Age-stratified analyses revealed also associations with high coworker support (1.2, 1.0-1.4) and strenuous leisure-time physical exercise (1.2, 1.0-1.4) in those aged 30-44 and low alcohol consumption (1.8, 1.2-2.6) in the age-group 45-64. Several potentially modifiable factors related to health, work, and lifestyle were associated with good WA among occupationally active subjects with MSP. © 2015 the Nordic Societies of Public Health.

The genetic polymorphism of merozoite surface protein-1 in Plasmodium falciparum isolates from Aceh province, Indonesia

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Jamil, K. F.; Supargiyono, S.; Syafruddin, D.; Pratama, N.; Silvy, S.

2018-03-01

An estimated of 3.3 million Indonesian population were infected with malaria. However, extensive genetic polymorphism of the field isolates msp-1 of P. falciparum represents a major obstacle for the development of malaria treatment. The aim of this study was to investigate the genetic diversity of msp-1 genotype in field isolates of P. falciparum collected in Aceh Province. A total of 90 patients with malaria (+) were selected from eleven district hospitals in Aceh from 2013-2015. Data were collected by anamnesis, complete physical examination and laboratory tests for msp-1. All protocols to diagnose malaria followed the WHO 2010 guideline. All samples were stored in Eijkman Biology Molecular Institute, Jakarta. Among 90 samples, 57.7% were male, and 42.3% were female with the most cases found between 21-30 years old. From the allele typing analysis of P. falciparum from Aceh; K1, MAD20, and RO33 allele types were identified. MAD20 type was the highest allele found in this study (57.9%). It was found in single and mixed infection. A moderate level of the mixed allele was also observed.

A mutation in human VAP-B--MSP domain, present in ALS patients, affects the interaction with other cellular proteins.

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Mitne-Neto, M; Ramos, C R R; Pimenta, D C; Luz, J S; Nishimura, A L; Gonzales, F A; Oliveira, C C; Zatz, M

2007-09-01

Amyotrophic Lateral Sclerosis (ALS) is the most common adult-onset Motor Neuron Disease (MND), characterized by motor neurons death in the cortex, brainstem and spinal cord. Ten loci linked to Familial ALS have been mapped. ALS8 is caused by a substitution of a proline by a serine in the Vesicle-Associated Membrane Protein-Associated protein-B/C (VAP-B/C). VAP-B belongs to a highly conserved family of proteins implicated in Endoplasmic Reticulum-Golgi and intra-Golgi transport and microtubules stabilization. Previous studies demonstrated that the P56S mutation disrupts the subcellular localization of VAP-B and that this position would be essential for Unfolded Protein Response (UPR) induced by VAP-B. In the present work we expressed and purified recombinant wild-type and P56S mutant VAP-B-MSP domain for the analysis of its interactions with other cellular proteins. Our findings suggest that the P56S mutation may lead to a less stable interaction of this endoplasmic reticulum protein with at least two other proteins: tubulin and GAPDH. These two proteins have been previously related to other forms of neurodegenerative diseases and are potential key points to understand ALS8 pathogenesis and other forms of MND. Understanding the role of these protein interactions may help the treatment of this devastating disease in the future.

The characterization and manipulation of the bacterial microbiome of the Rocky Mountain wood tick, Dermacentor andersoni.

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Clayton, Katie A; Gall, Cory A; Mason, Katheen L; Scoles, Glen A; Brayton, Kelly A

2015-12-10

In North America, ticks are the most economically impactful vectors of human and animal pathogens. The Rocky Mountain wood tick, Dermacentor andersoni (Acari: Ixodidae), transmits Rickettsia rickettsii and Anaplasma marginale to humans and cattle, respectively. In recent years, studies have shown that symbiotic organisms are involved in a number of biochemical and physiological functions. Characterizing the bacterial microbiome of D. andersoni is a pivotal step towards understanding symbiont-host interactions. In this study, we have shown by high-throughput sequence analysis that the composition of endosymbionts in the midgut and salivary glands in adult ticks is dynamic over three generations. Four Proteobacteria genera, Rickettsia, Francisella, Arsenophonus, and Acinetobacter, were identified as predominant symbionts in these two tissues. Exposure to therapeutic doses of the broad-spectrum antibiotic, oxytetracycline, affected both proportions of predominant genera and significantly reduced reproductive fitness. Additionally, Acinetobacter, a free-living ubiquitous microbe, invaded the bacterial microbiome at different proportions based on antibiotic treatment status suggesting that microbiome composition may have a role in susceptibility to environmental contaminants. This study characterized the bacterial microbiome in D. andersoni and determined the generational variability within this tick. Furthermore, this study confirmed that microbiome manipulation is associated with tick fitness and may be a potential method for biocontrol.

Genetic Diversity of Plasmodium falciparum Populations in Malaria Declining Areas of Sabah, East Malaysia.

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Mohd Ridzuan Mohd Abd Razak

Full Text Available Malaysia has a national goal to eliminate malaria by 2020. Understanding the genetic diversity of malaria parasites in residual transmission foci can provide invaluable information which may inform the intervention strategies used to reach elimination targets. This study was conducted to determine the genetic diversity level of P. falciparum isolates in malaria residual foci areas of Sabah. Malaria active case detection was conducted in Kalabakan and Kota Marudu. All individuals in the study sites were screened for malaria infection by rapid diagnostic test. Blood from P. falciparum-infected individuals were collected on filter paper prior to DNA extraction. Genotyping was performed using merozoite surface protein-1 (MSP-1, merozoite surface protein-2 (MSP-2, glutamate rich protein (GLURP and 10 neutral microsatellite loci markers. The size of alleles, multiplicity of infection (MOI, mean number of alleles (Na, expected heterozygosity (He, linkage disequilibrium (LD and genetic differentiation (FST were determined. In Kalabakan, the MSP-1 and MSP-2 alleles were predominantly K1 and FC27 family types, respectively. The GLURP genotype VI (751-800 bp was predominant. The MOI for MSP-1 and MSP-2 were 1.65 and 1.20, respectively. The Na per microsatellite locus was 1.70. The He values for MSP-1, MSP-2, GLURP and neutral microsatellites were 0.17, 0.37, 0.70 and 0.33, respectively. In Kota Marudu, the MSP-1 and MSP-2 alleles were predominantly MAD20 and 3D7 family types, respectively. The GLURP genotype IV (651-700 bp was predominant. The MOI for both MSP-1 and MSP-2 was 1.05. The Na per microsatellite locus was 3.60. The He values for MSP-1, MSP-2, GLURP and neutral microsatellites were 0.24, 0.25, 0.69 and 0.30, respectively. A significant LD was observed in Kalabakan (0.495, p<0.01 and Kota Marudu P. falciparum populations (0.601, p<0.01. High genetic differentiation between Kalabakan and Kota Marudu P. falciparum populations was observed (FST = 0

Prevalence of Babesia canis, Borrelia burgdorferi sensu lato, and Anaplasma phagocytophilum in hard ticks collected from meadows of Lubelskie Voivodship (eastern Poland

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Dzięgiel Beata

2014-03-01

Full Text Available The aim of the study was to assess the distribution of Anaplasma phagocytophilum, Borrelia burgdorferi sensu lato, and Babesia canis in adult females and males of Ixodes ricinus and Dermacentor reticulatus ticks, inhabiting meadows near large forest complexes throughout the Lubelskie Voivodship (eastern region of Poland. Ticks were collected using the flagging method. Among 720 ticks collected, 506 were identified as D. reticulatus, and 214 as I. ricinus. DNA of B. canis and B. burgdorferi s.l. was detected in 21.3% and 0.6% of D. reticulatus ticks, respectively. In I. ricinus ticks, DNA specific to B. burgdorferi s.l. and A. phagocytophilum was detected in 5.6% and 10.3%, respectively. Co-infections of B. burgdorferi s.l. and A. phagocytophilum were found in two I. ricinus ticks. These results indicate that the Lublin region is an area at risk of tick-borne diseases of humans and animals, which must be considered in clinical practice.

Infection of Anaplasma phagocytophilum and Ehrlichia spp. in Opossums and Dogs in Campeche, Mexico: The Role of Tick Infestation

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Edgar Rojero-Vázquez

2017-12-01

Full Text Available In recent years, some tick-borne diseases such as anaplasmosis and ehrlichiosis became widespread worldwide, threatening the health of humans, domestic animals and wildlife. The aims of this study were to determine the presence of Anaplasma phagocytophilum, Ehrlichia canis, and Ehrlichia chaffeensis in 102 opossums (Didelphis spp. and 44 owned free-ranging dogs in southeastern Mexico using a specific polymerase chain reaction (PCR. A. phagocytophilum was detected in opossums and dogs with a prevalence of 3 and 27%, respectively. E. canis was only present in 7% of dogs, while we didn't detect E. chaffeensis in any host. We report the first evidence of infections of A. phagocytophilum in Didelphis virginiana and D. marsupialis in Mexico. The infection rates and patterns we found of A. phagocytophilum suggest that dogs are more directly involved in the ecology of this pathogen than opossums. Despite the small prevalence found, our results are of public health concern because of the zoonotic capabilities of A. phagocytophilum, the high tick infestation rates found and because both opossums and free-ranging dogs can achieve high population densities in the region.

Dynamics of anti-MSP3 and Pfs230 antibody responses and multiplicity of infection in asymptomatic children from southern Ghana

DEFF Research Database (Denmark)

Amoah, Linda E; Acquah, Festus K; Ayanful-Torgby, Ruth

2018-01-01

BACKGROUND: During a Plasmodium infection, exposure of human host immune cells to both the asexual and the sexual stages of the parasite elicit immune responses. These responses may be protective and prevent the development of high parasitaemia and its associated clinical symptoms, or block...... the transmission of malaria to an uninfected person. This study aimed at examining the dynamics of naturally acquired immune responses against the asexual and sexual forms of Plasmodium falciparum as well as assessing differences in the multiplicity of infection (MOI) in asymptomatic Ghanaian children living...... 2015. Filter paper blood blots (DBS) as well as thick and thin blood smears were made from finger-pricked blood at each visit. Plasmodium falciparum parasite prevalence was determined by microscopy and PCR. Serum eluted from the DBS were used to assess anti-Pfs230 (sexual stage) and anti-MSP3 (asexual...

Prevalence of Ehrlichia canis, Anaplasma platys, Babesia canis vogeli, Hepatozoon canis, Bartonella vinsonii berkhoffii, and Rickettsia spp. in dogs from Grenada.

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Yabsley, Michael J; McKibben, John; Macpherson, Calum N; Cattan, Peggy F; Cherry, Natalie A; Hegarty, Barbara C; Breitschwerdt, Edward B; O'Connor, Tom; Chandrashekar, Ramaswamy; Paterson, Tara; Perea, Marta Lanza; Ball, Geoffrey; Friesen, Stanley; Goedde, Jill; Henderson, Brooke; Sylvester, Wayne

2008-02-14

To identify the tick-borne pathogens in dogs from Grenada, we conducted a serologic survey for Ehrlichia canis in 2004 (104 dogs) and a comprehensive serologic and molecular survey for a variety of tick-borne pathogens in 2006 (73 dogs). In 2004 and 2006, 44 and 32 dogs (42.3% and 43.8%) were seropositive for E. canis, respectively. In 2006, several tick-borne pathogens were identified by serology and PCR. DNA of E. canis, Anaplasma platys, Babesia canis vogeli, Hepatozoon canis, and Bartonella sp. were identified in 18 (24.7%), 14 (19.2%), 5 (7%), 5 (7%), and 1 (1.4%) dogs, respectively. Six (8.2%) dogs were seropositive for Bartonella vinsonii subsp. berkhoffii. All dogs were seronegative and PCR-negative for Rickettsia spp. Coinfection with two or three pathogens was observed in eight dogs. Partial 16S rRNA E. canis and A. platys sequences were identical to sequences in GenBank. Partial 18S rRNA gene sequences from the Grenadian H. canis were identical to each other and had one possible mismatch (ambiguous base) from H. canis detected from Spain and Brazil. Grenadian B. c. vogeli sequences were identical to B. c. vogeli from Brazil and Japan. All of the detected pathogens are transmitted, or suspected to be transmitted, by Rhipicephalus sanguineus. Results of this study indicate that dogs from Grenada are infected with multiple tick-borne pathogens; therefore, tick-borne diseases should be included as differentials for dogs exhibiting thrombocytopenia, leukopenia, fever, or lethargy. One pathogen, E. canis, is also of potential public health significance.

A Search for Millisecond-pulsar Radio Emission from the Faint Quiescent Soft X-Ray Transient 1H 1905+000

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Mikhailov, K.; Van Leeuwen, J. [Anton Pannekoek Institute for Astronomy, University of Amsterdam, Science Park 904, P.O. Box 94249, 1090 GE Amsterdam (Netherlands); Jonker, P. G., E-mail: K.Mikhailov@uva.nl [SRON, the Netherlands Institute for Space Research, Sorbonnelaan 2, 3584 CA, Utrecht (Netherlands)

2017-05-01

Transitional millisecond pulsars (tMSPs) switch between an accretion-powered state without radio pulsations and a rotation-powered state with radio pulsations. In the former state, tMSPs are X-ray bright, while in the latter state, they are X-ray dim. Soft X-ray transients (SXTs) undergo similar switches in X-ray, between “high” states with bright X-ray outbursts and “low” states of quiescence. The upper limit on the quiescent X-ray luminosity of SXT 1H 1905+000 suggests that its luminosity might be similar to that of the known tMSPs. A detection of radio pulsations would link SXTs more strongly with tMSPs; and thus, e.g., put stricter constraints on tMSP transitional timescales through the connection with the well-known SXT periods of quiescence. A nondetection allows us, based on the telescope sensitivity, to estimate how likely these sources are to pulsate in radio. Over a 10-year span, 2006–2015, we carried out targeted radio observations at 400/800 MHz with Arecibo, and searched for radio pulsations from the quiescent SXT 1H 1905+000. None of the observations have revealed radio pulsations from the targeted SXT. For a 1 ms pulsar, our flux density upper limit is 10.3 μ Jy. At an assumed distance of 10 kpc this translates to a pseudo-luminosity upper limit of 1.0 mJy kpc{sup 2}, which makes our search complete to ∼85% of the known MSP population. Given the high sensitivity, and the generally large beaming fraction of millisecond pulsars, we conclude that SXT 1H 1905+000 is unlikely to emit in radio as a tMSP.

Interaction between estrogen receptor and retinol-binding protein-4 polymorphisms as a tool for the selection of prolific pigs

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Iara Denise Vasconcellos Gonçalves

2008-01-01

Full Text Available The aim of the present study was to investigate the association of the estrogen receptor (ER-PvuII and retinol-binding protein 4 (RBP4-MspI gene polymorphisms and their interactions with prolificacy in a commercial synthetic pig line reared in Brazil. A total of 10,374 piglet records from 218 sows and 817 litters were used for litter size analysis. Only females with three or four farrowings were included in the analysis. The mean litter size ranged from 5.0 to 19.5 piglets. DNA was extracted from leukocytes by a standard method, and ER-PvuII and RBP4-MspI polymorphisms were characterized by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP technique. The association between alleles or genotypes and reproductive performance was analyzed using a general linear model including the interaction between the ER-PvuII and RBP4-MspI genotypes. For the ER-PvuII gene, the allele frequencies of allele A and allele B were 0.56 and 0.44, respectively. For the RBP4-MspI gene, the frequencies of alleles A1 and A2 were 0.29 and 0.71, respectively. The total number of piglets born (TNB, born alive (NBA, or number of mummies and stillborn piglets (NMUM and NSB per litter did not differ between the various ER-PvuII and RBP4-MspI genotypes. However, when the ER-PvuII and RBP4-MspI genotypes were considered together in each sow, TNB and NBA were 1.4 (p = 0.0026 and 0.9 (p = 0.019 higher in AA/A1 and AB/A1 animals, respectively, than in AA/A2 and BB/A1 animals. Likewise, TNB and NBA were 0.9 (p = 0.0258 and 0.8 (p = 0.0168 higher in BB/A2 and AB/A2 sows, respectively, than in AA/A2 and BB/A1 animals, but no difference was observed compared to AA/A1 and AB/A1 animals. The results showed larger litter sizes (TNB and NBA for sows carrying the ER-PvuII allele A and the RBP4-MspI genotype A1, and for animals carrying the ER-PvuII allele B and the RBP4-MspI genotype A2. In conclusion, the interaction between genotypes ER-PvuII and RBP4-MspI is

The high accuracy data processing system of laser interferometry signals based on MSP430

Science.gov (United States)

Qi, Yong-yue; Lin, Yu-chi; Zhao, Mei-rong

2009-07-01

Generally speaking there are two orthogonal signals used in single-frequency laser interferometer for differentiating direction and electronic subdivision. However there usually exist three errors with the interferential signals: zero offsets error, unequal amplitude error and quadrature phase shift error. These three errors have a serious impact on subdivision precision. Based on Heydemann error compensation algorithm, it is proposed to achieve compensation of the three errors. Due to complicated operation of the Heydemann mode, a improved arithmetic is advanced to decrease the calculating time effectively in accordance with the special characteristic that only one item of data will be changed in each fitting algorithm operation. Then a real-time and dynamic compensatory circuit is designed. Taking microchip MSP430 as the core of hardware system, two input signals with the three errors are turned into digital quantity by the AD7862. After data processing in line with improved arithmetic, two ideal signals without errors are output by the AD7225. At the same time two original signals are turned into relevant square wave and imported to the differentiating direction circuit. The impulse exported from the distinguishing direction circuit is counted by the timer of the microchip. According to the number of the pulse and the soft subdivision the final result is showed by LED. The arithmetic and the circuit are adopted to test the capability of a laser interferometer with 8 times optical path difference and the measuring accuracy of 12-14nm is achieved.

Geographic Variation in Maternal Smoking during Pregnancy in the Missouri Adolescent Female Twin Study (MOAFTS)

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Lian, Min; Madden, Pamela A.; Lynskey, Michael T.; Colditz, Graham A.; Lessov-Schlaggar, Christina N.; Schootman, Mario; Heath, Andrew C.

2016-01-01

Objective Despite well-known adverse health effects of maternal smoking during pregnancy (MSP), it is still unclear if MSP varies geographically and if neighborhood socioeconomic deprivation (SED) plays an important role in MSP. This study aims to investigate small-area geographic variation in MSP and examine the association of SED with MSP. Methods The Missouri Adolescent Female Twin Study (MOAFTS) is a cohort study of female like-sex twins born in Missouri to Missouri-resident parents during 1975–1985. Biological mothers completed a baseline interview in 1995–1998 and reported MSP with the twins. Residential address of the mother at birth was geocoded. We developed a census tract-level SED index using a common factor approach based on 21 area-level socioeconomic variables from the 1980 Census data. Multilevel logistic regressions estimated geographic heterogeneity (random effect) in MSP and the odds ratios (ORs, fixed effects) of neighborhood SED associated with MSP. Results Of 1658 MOAFTS mothers, 35.2% reported any MSP and 21.9% reported MSP beyond the first trimester. Neighborhood SED was associated with any MSP (the highest vs. the lowest quartile: OR = 1.90, 95% confidence interval [CI] = 1.40–2.57, Ptrendalcohol use, and parents’ cohabitation), neighborhood SED was not associated with MSP, but geographic variation still persisted in MSP (variance = 0.41, P = 0.003) and in MSP beyond the first trimester (variance = 0.82, P<0.001). Conclusions Neighborhood SED was associated with MSP in unadjusted analyses but this association could be explained by individual socioeconomic conditions. Nonetheless, significant geographic variation in MSP persisted and was not accounted for by differences in neighborhood SED. To develop effective interventions to reduce MSP, further studies are necessary to explore underlying reasons for its geographic variation. PMID:27100091

Babesia spp. and Anaplasma phagocytophilum in questing ticks, ticks parasitizing rodents and the parasitized rodents--analyzing the host-pathogen-vector interface in a metropolitan area.

Science.gov (United States)

Silaghi, Cornelia; Woll, Dietlinde; Hamel, Dietmar; Pfister, Kurt; Mahling, Monia; Pfeffer, Martin

2012-09-05

The aims of this study were to evaluate the host-tick-pathogen interface of Babesia spp. and Anaplasma phagocytophilum in restored areas in both questing and host-attached Ixodes ricinus and Dermacentor reticulatus and their small mammalian hosts. Questing ticks were collected from 5 sites within the city of Leipzig, Germany, in 2009. Small mammals were trapped at 3 of the 5 sites during 2010 and 2011. DNA extracts of questing and host-attached I. ricinus and D. reticulatus and of several tissue types of small mammals (the majority bank voles and yellow-necked mice), were investigated by PCR followed by sequencing for the occurrence of DNA of Babesia spp. and by real-time PCR for A. phagocytophilum. A selected number of samples positive for A. phagocytophilum were further investigated for variants of the partial 16S rRNA gene. Co-infection with Rickettsia spp. in the questing ticks was additionally investigated. 4.1% of questing I. ricinus ticks, but no D. reticulatus, were positive for Babesia sp. and 8.7% of I. ricinus for A. phagocytophilum. Sequencing revealed B. microti, B. capreoli and Babesia spp. EU1 in Leipzig and sequence analysis of the partial 16S RNA gene of A. phagocytophilum revealed variants either rarely reported in human cases or associated with cervid hosts. The statistical analysis revealed significantly less ticks infected with A. phagocytophilum in a city park in Leipzig as compared to the other sampling sites. A. phagocytophilum-DNA was detected in 2 bank voles, DNA of B. microti in 1 striped field-mouse and of Babesia sp. EU1 in the skin tissue of a mole. Co-infections were detected. Our results show the involvement of small mammals in the natural endemic cycles of tick-borne pathogens. A more thorough understanding of the interactions of ticks, pathogens and hosts is the essential basis for effective preventive control measures.

TWRS Privatization Phase 1 Master Site Plan

International Nuclear Information System (INIS)

PARAZIN, R.J.

1999-01-01

The U.S. Department of Energy (DOE) has chosen to accomplish the Tank Waste Remediation System disposal mission via privatization. The disposal mission has been divided into two privatization phases. Phase I, a 'proof of concept' phase, will establish and demonstrate the technical, commercial, and procurement capabilities necessary far privatization to proceed. Once established, privatization will be expanded in the form of a second phase (Phase II) to dispose of the remainder of the tank waste. In conjunction with preparation of the Tank Waste Remediation System (TWRS) Privatization Request for Proposals (RFP)(RL, 1996), a location was selected for the Phase I demonstration facilities (Shord, 1996). The location selected was the area previously developed and characterized for the Grout Disposal Site, adjoining the 200 East Area. The site is of sufficient size for a Private Contractor (PC) to carry out pretreatment, immobilization, and vitrification operations and possesses the required characteristics (e.g., close to feed tanks) to best facilitate the Phase I operations. This overall long-range Master Site Plan (MSP) has been developed to establish a ''baseline'' for the (TWRS) Privatization Phase I (TPPI) PC Site. The MSP depicts the planned layout for the PC Site along with various interfaces between the site and other Hanford utilities and functions. The complete integration of TPPl MSP with overall Hanford Site planning process will assist in establishing the PC site and the necessary priorities to meet the Hanford cleanup mission. The MSP has been developed systematically into a comprehensive, safe, flexible, logical and cost-effective plan. The general philosophy behind the preparation of a MSP for the TPPl program is that it will serve as a single source documentation of the planning for the development of the TPPl complex. The effort will plan temporary and permanent land use, utilities, and traffic flow for the overall program. It will identify needs

The unmet needs of Aboriginal Australians with musculoskeletal pain: A mixed method systematic review.

Science.gov (United States)

Lin, Ivan B; Bunzli, Samantha; Mak, Donna B; Green, Charmaine; Goucke, Roger; Coffin, Juli; O'Sullivan, Peter B

2017-12-15

Musculoskeletal pain (MSP) conditions are the biggest cause of disability and internationally, Indigenous peoples experience a higher burden. There are conflicting reports about Aboriginal Australians and MSP. We conducted a systematic review to describe the prevalence, associated factors, impacts, care access, health care experiences, and factors associated with MSP among Aboriginal Australians. A systematic search of quantitative and qualitative scientific and grey literature (PROSPERO number: CRD42016038342). Articles were appraised using the Mixed Methods Appraisal Tool. Due to study heterogeneity a narrative synthesis was conducted. Of 536 articles identified, 18 were included (14 quantitative, 4 qualitative), of high (n=11), medium (n=2) and low (n=5) quality. Prevalences of MSP in Aboriginal populations were similar to or slightly higher than the non-Aboriginal population (prevalence rate ratio 1.1 for back pain, 1.2-1.5 for osteoarthritis (OA), 1.0-2.0 for rheumatoid arthritis). Aboriginal people accessed primary care for knee or hip OA at around half the rate of non-Aboriginal people, and were less than half as likely to have knee or hip replacement surgery. Communication difficulties with health practitioners were the main reason why Aboriginal people with MSP choose not to access care. No articles reported interventions. Findings provide preliminary evidence of an increased MSP burden amongst Aboriginal Australians and, particularly for OA, a mismatch between the disease burden and access to health care. To increase accessibility, health services should initially focus on improving Aboriginal patients' experiences of care, in particular by improving patient-practitioner communication. Implications for care and research are outlined. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.

Plasmodium vivax merozoite surface protein-3 alpha: a high-resolution marker for genetic diversity studies.

Science.gov (United States)

Prajapati, Surendra Kumar; Joshi, Hema; Valecha, Neena

2010-06-01

Malaria, an ancient human infectious disease caused by five species of Plasmodium, among them Plasmodium vivax is the most widespread human malaria species and causes huge morbidity to its host. Identification of genetic marker to resolve higher genetic diversity for an ancient origin organism is a crucial task. We have analyzed genetic diversity of P. vivax field isolates using highly polymorphic antigen gene merozoite surface protein-3 alpha (msp-3 alpha) and assessed its suitability as high-resolution genetic marker for population genetic studies. 27 P. vivax field isolates collected during chloroquine therapeutic efficacy study at Chennai were analyzed for genetic diversity. PCR-RFLP was employed to assess the genetic variations using highly polymorphic antigen gene msp-3 alpha. We observed three distinct PCR alleles at msp-3 alpha, and among them allele A showed significantly high frequency (53%, chi2 = 8.22, p = 0.001). PCR-RFLP analysis revealed 14 and 17 distinct RFLP patterns for Hha1 and Alu1 enzymes respectively. Further, RFLP analysis revealed that allele A at msp-3 alpha is more diverse in the population compared with allele B and C. Combining Hha1 and Alu1 RFLP patterns revealed 21 distinct genotypes among 22 isolates reflects higher diversity resolution power of msp-3 alpha in the field isolates. P. vivax isolates from Chennai region revealed substantial amount of genetic diversity and comparison of allelic diversity with other antigen genes and microsatellites suggesting that msp-3 alpha could be a high-resolution marker for genetic diversity studies among P. vivax field isolates.

Identification and characterization of polymorphisms at the HSA a1-acid glycoprotein (ORM* gene locus in Caucasians

Directory of Open Access Journals (Sweden)

Owczarek Catherine M.

2002-01-01

Full Text Available Human alpha1-acid glycoprotein (AGP or orosomucoid (ORM is a major acute phase protein that is thought to play a crucial role in maintaining homeostasis. Human AGP is the product of a cluster of at least two adjacent genes located on HSA chromosome 9. Using a range of restriction endonucleases we have investigated DNA variation at the locus encoding the AGP genes in a panel of healthy Caucasians. Polymorphisms were identified using BamHI, EcoRI, BglII, PvuII, HindIII, TaqI and MspI. Non-random associations were found between the BamHI, EcoRI, BglII RFLPs. The RFLPs detected with PvuII, TaqI and MspI were all located in exon 6 of both AGP genes. The duplication of an AGP gene was observed in 11% of the indiviuals studied and was in linkage disequilibrium with the TaqI RFLP. The identification and characterization of these polymorphisms will prove useful for other population and forensic studies.

Molecular Survey on Rickettsia spp., Anaplasma phagocytophilum, Borrelia burgdorferi Sensu Lato, and Babesia spp. in Ixodes ricinus Ticks Infesting Dogs in Central Italy.

Science.gov (United States)

Morganti, Giulia; Gavaudan, Stefano; Canonico, Cristina; Ravagnan, Silvia; Olivieri, Emanuela; Diaferia, Manuela; Marenzoni, Maria Luisa; Antognoni, Maria Teresa; Capelli, Gioia; Silaghi, Cornelia; Veronesi, Fabrizia

2017-11-01

Dogs are a common feeding hosts for Ixodes ricinus and may act as reservoir hosts for zoonotic tick-borne pathogens (TBPs) and as carriers of infected ticks into human settings. The aim of this work was to evaluate the presence of several selected TBPs of significant public health concern by molecular methods in I. ricinus recovered from dogs living in urban and suburban settings in central Italy. A total of 212 I. ricinus specimens were collected from the coat of domestic dogs. DNA was extracted from each specimen individually and tested for Rickettsia spp., Borrelia burgdorferi sensu lato, Babesia spp., and Anaplasma phagocytophilum, using real-time and conventional PCR protocols, followed by sequencing. Sixty-one ticks (28.8%) tested positive for TBPs; 57 samples were infected by one pathogen, while four showed coinfections. Rickettsia spp. was detected in 39 specimens (18.4%), of which 32 were identified as Rickettsia monacensis and seven as Rickettsia helvetica. Twenty-two samples (10.4%) tested positive for A. phagocytophilum; Borrelia lusitaniae and Borrelia afzelii were detected in two specimens and one specimen, respectively. One tick (0.5%) was found to be positive for Babesia venatorum (EU1). Our findings reveal the significant exposure of dogs to TBPs of public health concern and provide data on the role of dogs in the circulation of I. ricinus-borne pathogens in central Italy.

Babesia spp. and Anaplasma phagocytophilum in questing ticks, ticks parasitizing rodents and the parasitized rodents – Analyzing the host-pathogen-vector interface in a metropolitan area

Directory of Open Access Journals (Sweden)

Silaghi Cornelia

2012-09-01

Full Text Available Abstract Background The aims of this study were to evaluate the host-tick-pathogen interface of Babesia spp. and Anaplasma phagocytophilum in restored areas in both questing and host-attached Ixodes ricinus and Dermacentor reticulatus and their small mammalian hosts. Methods Questing ticks were collected from 5 sites within the city of Leipzig, Germany, in 2009. Small mammals were trapped at 3 of the 5 sites during 2010 and 2011. DNA extracts of questing and host-attached I. ricinus and D. reticulatus and of several tissue types of small mammals (the majority bank voles and yellow-necked mice, were investigated by PCR followed by sequencing for the occurrence of DNA of Babesia spp. and by real-time PCR for A. phagocytophilum. A selected number of samples positive for A. phagocytophilum were further investigated for variants of the partial 16S rRNA gene. Co-infection with Rickettsia spp. in the questing ticks was additionally investigated. Results 4.1% of questing I. ricinus ticks, but no D. reticulatus, were positive for Babesia sp. and 8.7% of I. ricinus for A. phagocytophilum. Sequencing revealed B. microti, B. capreoli and Babesia spp. EU1 in Leipzig and sequence analysis of the partial 16S RNA gene of A. phagocytophilum revealed variants either rarely reported in human cases or associated with cervid hosts. The statistical analysis revealed significantly less ticks infected with A. phagocytophilum in a city park in Leipzig as compared to the other sampling sites. A. phagocytophilum-DNA was detected in 2 bank voles, DNA of B. microti in 1 striped field-mouse and of Babesia sp. EU1 in the skin tissue of a mole. Co-infections were detected. Conclusion Our results show the involvement of small mammals in the natural endemic cycles of tick-borne pathogens. A more thorough understanding of the interactions of ticks, pathogens and hosts is the essential basis for effective preventive control measures.