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Simian malaria in the Brazilian Atlantic forest: first description of natural infection of capuchin monkeys (Cebinae subfamily) by Plasmodium simium
Malaria Journal volume 14, Article number: 81 (2015)
In Brazil, two species of Plasmodium have been described infecting non-human primates, Plasmodium brasilianum and Plasmodium simium. These species are morphologically, genetically and immunologically indistinguishable from the human Plasmodium malariae and Plasmodium vivax parasites, respectively. Plasmodium simium has been observed naturally infecting monkeys of the genera Alouatta and Brachyteles in a restricted area of the Atlantic Forest in the south and southeast regions of Brazil. However, its reported geographical distribution and the diversity of its vertebrate hosts may be underestimated, since available data were largely based on analyses by microscopic examination of peripheral blood, a method with limited sensitivity, considering the potential sub-patent feature of these infections. The present study describes, for the first time, the natural infection of P. simium in capuchin monkeys from the Brazilian Atlantic Forest.
Blood samples from 30 non-human primates belonging to nine species kept in the Primate Centre of Rio de Janeiro were collected. Fragments of spleen and liver from one dead monkey found in the neighborhoods of the Primate Centre were also analysed. Molecular diagnosis was performed by nested PCR (18SSU rRNA) and the amplified fragment was sequenced.
Thirty per cent of the captive animals were infected with P. simium and/or P. brasilianum. The dead monkey tested positive for DNA of P. simium. For the first time, Cebinae primates (two specimens of genus Cebus and two of genus Sapajos) were found naturally infected by P. simium. The infection was confirmed by sequencing a small fragment of 18SSU rRNA.
The results highlight the possibility of infection by P. simium in other species of non-human primates whose impact could be significant for the malaria epidemiology among non-human primates and, if it becomes clear that this P. simium is able to infect monkeys and, eventually, man, also for the maintenance of transmission of human malaria in the context of a zoonosis in areas under influence of the Atlantic Forest.
Malaria, one of the major public health problems in the world, is a mosquito-borne disease caused by parasites of the genus Plasmodium that affects mammals, reptiles and birds . Plasmodium species causing infection of non-human primates are of great interest because they may be naturally or accidentally transmitted to humans [2-7]. In Brazil, a country that holds the largest species diversified simian fauna of the planet, only two simian Plasmodium species have been found infecting non-human primates: Plasmodium brasilianum and Plasmodium simium. These species are morphologically, genetically and immunologically indistinguishable from the human malaria parasites Plasmodium malariae and Plasmodium vivax, respectively [8-11]. Both, P. brasilianum and P. simium are infective to human [2,12].
Plasmodium brasilianum has a wide distribution in Central and South America, where it has been found in Brazil, Colombia, Venezuela, Panama, and Peru naturally infecting a large number of species belonging to all families of New World non-human primates, i.e., Aotidae, Atelidae, Cebidae, and Pitheciidae families [9,13]. In Brazil, excepting the arid portions in the northeast and savannah in the southeast, the territory range of P. brasilianum includes all regions and overlaps that of P. simium, which is restricted to the Atlantic Forest in the south and southeast . Contrarily to P. brasilianum, P. simium has been detected infecting only two Atelidae genera: Alouatta (howler monkeys) and Brachyteles (woolly spider monkeys) [9,14,15]. However, most of the previous simian malaria surveys performed in the Atlantic Forest were based on microscopy of blood smears . Given to the great diversity of primates in the Brazilian Atlantic Forest it is expected that, with the use of more sensitive diagnostic methods, the determination of the real rate of infection in monkeys and, eventually, the expansion of the list of potential hosts would be possible.
According to the National Malaria Control Program (PNCM), Secretary for Health Surveillance (SVS), Ministry of Health, from January 2006 to December 2013, 8,410 cases of malaria were reported in extra-Amazon region, 1,068 of them being autochthonous . In 2013, from the 827 registered extra-Amazonian cases, 10.6% (88 cases) were due to autochthonous transmission, corresponding to 0.05% of the total cases in Brazil . In south and southeastern Brazil, human malaria transmission has been essentially eliminated or has been reported during scattered outbreaks of introduced cases, due to imported ones, for more than four decades [17-19]. However, few autochthonous human malaria cases and outbreaks have been reported associated with the Atlantic Forest environment, particularly in its mountain valleys in southeast Brazil [9,17,19,20]. In such environments, malaria transmission is supported by the bromeliad mosquito Anopheles (Kerteszia) cruzii, which has been proved to be the natural vector of both human and simian malaria in south and southeast Brazil . Therefore, Deane and colleagues in the 1990s and others [8,9,15,17,21] have suggested that human malaria in the Atlantic Forest could be a zoonosis where non-human primates are the reservoirs.
Autochthonous human malaria cases reported in the mountain valleys of the Atlantic Forest environment in Rio de Janeiro State motivated the search for natural simian plasmodial infections in this region . The present study describes for the first time the natural infection of P. simium in the subfamily Cebinae by using molecular approaches.
Origin of primate specimens
Animals used in this study were captive primates from the Primate Centre of Rio de Janeiro (CPRJ). The CPRJ (IBAMA register number 458460) is a unit for wild monkey protection and is located in the municipality of Guapimirim, on the Serra dos Órgãos slopes (Figure 1), in an area under influence of the Atlantic Forest, about l00 km from the city of Rio de Janeiro. Serra dos Órgãos is part of the large coastal mountain chain in southeast Brazil named Serra do Mar. One wild animal, identified as an adult female of Alouatta guariba clamitans, was found dead nearby the CPRJ in December 2013 and an autopsy was performed, and spleen and liver fragments were included in the analysis. The present study was conduced in response to a demand of the Health Secretary of Rio de Janeiro State, to the Centro de Pesquisa, Diagnóstico e Treinamento em Malária (CPD-Mal), Fiocruz, in the context of an investigation on the autochthonous human cases registered in the surroundings of the CPRJ, and it had, therefore, the authorization of the Fiocruz Human Ethical Committee. As the handling of monkeys was exclusively done by CPRJ technicians, Fiocruz Animal Ethics Committee (CEUA) agreed to the protocol for sample collection.
Samples collection and DNA extraction
A total of 30 captive primates were sampled (Additional file 1) in November 2011. Of these, 18 were female (62%); the average age was six years, median of seven years (1–14). The average weight was of 2,800 g (700–7,500 g). No animal was splenectomized at the time of sample collection. Approximately 2 ml of blood were taken in vacuum tubes containing EDTA by femoral venipuncture. For just a few individuals sedation with Quetamina® and Midazolan was required. The samples collection was performed by the Med Vet Alcides Pissinatti, the blood samples were frozen and send to the Malaria laboratory of the CPqRR, Fiocruz.
DNA extraction from blood samples was made using the Gentra Puregene Blood kit (Qiagen, Venlo, The Netherlands) according to manufacturer’s recommendations. Fragments of spleen and liver from the dead monkey were used for DNA extraction using the Gentra Puregene kit (Qiagen, Venlo, The Netherlands) according to manufacturer’s recommendations. The DNA was eluted in a 100 μL volume and stored at −20°C until it was used.
The samples were subjected to nested-PCR  using primers for identification of human Plasmodium species, targeting the small subunit of 18S ribosomal RNA gene (18SSU rRNA). Briefly, all PCR reactions were performed in 20 μL volumes containing 250 μM each oligonucleotide primer, 10 μL of Master Mix (Promega) (0.3 units of Taq Polymerase, 200 μM each deoxyribonucleotide triphosphates and 1.5 mM MgCl2) and 2 μL DNA. The PCR assays were performed using an automatic thermocycler (PTC-100TM v.7.0) (MJ Research Inc, USA) and the following cycling parameters were used: an initial denaturation at 95°C for 5 min followed by 24 cycles of annealing at 58°C for 2 min, extension at 72°C for 2 min and denaturation at 94°C for 1 min followed by a final annealing incubation at 58°C for 2 min and extension at 72°C for 2 min. The temperature was then reduced to 4°C until the samples were taken. The cycling parameters for the second round of PCR were the same as the first reaction, but instead 30 cycles of amplification were used.
To prevent cross-contamination, the DNA extraction and master mix preparation were performed in “parasite DNA-free rooms” distinct from each other. Furthermore, each of these separate areas has different sets of pipettes and all procedures were performed using plugged pipette tips. DNA extraction was performed twice at different days. Every PCR reaction had a negative control, where DNA has replaced by water and also positive controls for each pair of primers. The sources of genomic DNA samples that served as positive controls in the nested PCR assays are: i) P. falciparum DNA, strain 3D7 maintained in Malaria Laboratory (CPqRR-FIOCRUZ MINAS); (ii) DNA of patient with high parasitaemia for P. vivax and DNA of P. simium of a non-human primate with an acute infection and parasitaemia confirmed by optical microscopy (BL10); (iii) DNA of P. brasilianum of MR4 (Malaria Research and Reference Reagent Resource Center – ATCC, USA).
The amplified fragments were visualized in electrophoresis on 1.5% agarose gel dissolved in 1x TAE buffer (40 mM Tris-acetate, 1 mM EDTA) with 5 μg/mL ethidium bromide (Invitrogen) in a horizontal system (Bio-Rad) at 100 V for about 30 min. The gels were be examined under UV transilluminator (UVP - Bio-Doc System it) and were filed under digital system. Electrophoresis was performed in a room specific for amplified DNA, with appropriated sets of pipettes and plugged pipettes tips.
Sequencing of PCR-amplified DNA
For DNA sequencing, PCR products were purified using Purification Kit QIAquick (Qiagen) following manufacturer’s procedure. Around 3 ng of purified PCR products were amplified using 2.0 μM of each primer (forward or reverse of species-specific primers of second reaction) and 1 μL of Big Dye terminator kit in a program of: 96°C for 1 min, 35 cycles of 96°C for 15 sec, the temperature of primer annealing for 15 sec and 60°C for 15 sec. The fragments were precipitated using ammonium acetate, ressuspended in formamide HI-DI (Applied Biosystems) and electrophoretically separated in ABI 3730 DNA automatic sequencer.
The sequence data were analysed using the Blast program and the sequences aligned using the Clustal W and CAP3 programs in Bioedit package and also Muscle multiple alignment program . The tree was constructed using the maximum likelihood method with Tamura 3-parameter model  and 5,000 bootstrap replicates in MEGA 6.0 software .
Molecular diagnosis was performed in samples from 30 non-human primates from the CPRJ, and nine samples (30%) were found to be positive by nested-PCR: five samples were positive for P. brasilianum; three samples were positive for P. simium and one sample was positive for both P. simium and P. brasilianum (Figure 2). Negative controls never showed any amplified fragments and positive controls showed fragments with the expected sizes (Figure 2). Plasmodium simium was identified in Sapajus xanthosternos, Sapajus robustus and Cebus sp.; and P. brasilianum was detected in Sapajus xanthosternos, Sapajus robustus, Callicebus personatus, Aotus nigriceps and Alouatta g. clamitans (see Additional file 1). The dead wild monkey found nearby the CPRJ, sample MP1943, an Alouatta g. clamitans, was also found positive for P. simium by using DNA obtained from the spleen fragment and, with less intensity of amplification, from the liver fragment (Figure 3).
A species-specific fragment of 18SSU rRNA from P. simium identified in the molecular diagnosis was sequenced to confirm the infection for P. simium in monkeys from the Cebinae subfamily. Then the nucleotide sequence of 18SSU rRNA fragments from four infected animals (2324, 2559, 2503, and MP1943) identified here were compared to P. vivax and other Plasmodium species sequences available at Genbank (Figure 4). From the alignment, the high genetic similarity between P. simium and P. vivax was evident (one difference out of 86 nt). It was also able to discern between the P. vivax/P. simium from the other species of Plasmodium. In addition, this alignment was used to reconstruct the phylogenetic relationships of the simian and human Plasmodium. Plasmodium simium sequences clustered in a single branch. Moreover, all samples of P. simium are in the clade of P. vivax, reinforcing the genetic similarity between these parasites (Figure 5).
By molecular analysis of blood samples of 30 captive primates from a primate center in southeastern Brazil, 30% of the individuals were found infected with Plasmodium. Moreover, the dead monkey (Alouatta clamitans) found close to the CPRJ and also studied here showed infection by P. simium through detection of Plasmodium DNA in spleen and liver fragments. Interestingly, previous surveys based on microscopy of simian blood smears conducted in the same primate center (CPRJ) as well in other localities of the state of Rio de Janeiro failed to detect Plasmodium infections [26,27].
Although microscopic slides were not obtained, the presence of parasite DNA, even indicating possibly a low parasitaemia in the examined animals, points to the identification of a new natural host of P. simium: the capuchins monkeys. Capuchins monkeys are New World monkeys of the family Cebidae, subfamily Cebinae, that have the largest geographical distribution among the neotropical primates [28-32]. Prior to 2011, the subfamily Cebinae contained only a single genus, Cebus. However, recent studies have suggested the division of capuchin monkeys into two genera, Sapajus for robust, tufted capuchins and Cebus for untufted, gracile capuchins . Cebus and Sapajus have been described infected only by P. brasilianum [9,34]. However, this study describes that primates belonging to three species from both genera were found infected by P. simium. According to the literature, P. simium was described infecting only two species of genus Alouatta and one of Brachyteles primates [9,15]. Also, no evidence of experimental infection was obtained in splenectomized Cebus sp. inoculated with P. simium heavily parasitized blood sample . Based on microscopy of blood smears, the natural infection rates for P. brasilianum in capuchin monkeys are usually very low compared to other Cebidae species, such as squirrel monkeys, as well as Pitheciidae and Atelidae species [9,13]. Thus, the finding of P. simium DNA in blood samples of capuchin monkeys from Rio de Janeiro suggests that these primates possibly undergoes Plasmodium infections with very low or transient parasitaemia.
Although the prevalence of Cebidae monkeys found infected in the CPRJ, which is located inside the Atlantic Forest, is similar to those reported for other species of monkeys found to carry Plasmodium parasites in the region [9,15,21], it is still necessary to determine if captive monkeys were caught infected by Plasmodium or if the transmission has occurred at the CPRJ, facilitated by the simultaneous presence of the vector and infected monkeys. Plasmodium infection detected in one monkey born in CPRJ (monkey 2324), the naturally infected (dead) wild monkey in the neighborhoods of CPRJ and the existence of malaria vectors such as An. (Ker.) cruzii, Anopheles (Nyssorhynchus) albitarsis and Anopheles (Nyssorhynchus) aquasalis in the region may support the latter hypothesis [36,37]. In addition, the high proportion of monkeys found carrying antibodies against blood forms and sporozoites of P. brasilianum and P. simium may either reinforce the hypothesis of an increased transmission in the CPRJ neighborhoods or suggest that the prevalence of Plasmodium infection may be much higher than expected in this area of Atlantic Forest [9,15,21,38].
As all measures to avoid contamination have been undertaken here and the PCR reactions were very well controlled (see Methods) the results presented here can not result from cross-contamination in the laboratory. Therefore, the present list of P. simium-susceptible, neotropical, non-human primate species might be incomplete because of the usual nature of sub-patent infections along with the limited sensitivity of the diagnostic methods used, that were mainly based on analyses of peripheral blood microscopy. The findings showed here are strengthened by those of Duarte et al. who described the occurrence of antibodies against the P. vivax circumsporozoite surface protein (CSP) in Cebus sp. from Brazil . The presence of individuals from the Cebus genus carrrying antibodies against P. vivax DBPII, MSP119 and AMA1 has also been identified by our group in monkeys from different areas (unpublished data).
New World simian Plasmodium are very closely related to human Plasmodium; P. brasilianum and P. simium are similar to the human P. malariae and P. vivax parasites, respectively. The sequencing of a small fragment of the18SSU rRNA encoding gene showed a high genetic similarity between P. vivax and P. simium, as demonstrated by Tazi and Ayala . This high identity is also observed in the phylogenetic reconstruction of P. simium isolates studied here. It is important to notice that the small size of the sequenced fragment may hinder their ability to reproduce the phylogeny of the genus Plasmodium. Moreover, the high level of conservation in 18SSU rRNA among close species, such as P. vivax and P. simium, hamper the discussion if these species are distinct or not from each other. Many authors based on an analysis of different molecular targets, such as the conserved regions of the gene coding for the circumsporozoite surface protein (CSP) or mitochondrial genes have been suggesting the existence of two different species, P. simium and P. vivax, very closely related and their origin was through multiple host switches recently in the evolution [40-42].
The evolutionary history of P. vivax in the Americas is still very controversial. The most accepted hypothesis is that of post-Columbian origin, where P. vivax would have arrived with Europeans during the colonization of the Americas [43,44]. In the New World, several evidences suggest a recent introduction of P. vivax [43,45,46]. The evolutionary history of P. vivax in the Americas was studied by analysing the mitochondrial genome variability of these parasites. Many different genetic groups circulating in South America were found, and they have probably originated from independent introductions .
Despite the data presented here is not sufficient to support a deep discussion about this subject, the evidence collected so far suggest that P. simium is possibly a different species from P. vivax, based especially on some differences on the morphology and, more recently, polymorphisms in some genomic sequences. However, additional genomic information on P. simium is still needed to support this hypothesis.
Taken together, the data reinforce the hypothesis that non-human primates of Atlantic Forest, including species which have never be incriminated before, such as the capuchins monkeys, could be acting as a reservoir for Plasmodium. Once autochthonous human cases of malaria are reported in those regions, the presence of possible wild reservoirs may have important implications for public health, due to close contact between humans and monkeys in some parts of the Atlantic Forest environment [17,19].
The findings presented here might impact the knowledge on the epidemiology of the disease among human and non-human primates in the Brazilian Atlantic Forest. If it becomes clear that the same parasite can infect both humans and monkeys in this area, the presence of possible animal reservoirs may have important implications for public health  and areas of Atlantic Forest should be under strict surveillance for the possibility of a zoonotic pattern of malaria transmission. However, more studies are still needed to confirm this, particularly the detection of gametocytes in the monkeys and the identification of the same parasites circulating among the vectors and both hosts: human and simian. Due to the known close similarity between P. simium and P. vivax, the data reported here indicate that Cebus and Sapajus monkeys should have their susceptibility to P. vivax investigated in order to determine if they could represent new experimental models for studies with human Plasmodium, in addition to the Saimiri and Aotus neotropical monkeys recommended by the World Health Organization .
Perkins SL, Austin CC. Four new species of Plasmodium from New Guinea lizards: integrating morphology and molecules. J Parasitol. 2009;95:424–33.
Deane LM, Deane MP, Ferreira Neto J. Studies on transmission of simian malaria and on the natural infection of man with Plasmodium simium in Brazil. Bull World Health Organ. 1966;35:805–8.
Arruda ME, Nardini EH, Nussenzweig RS, Cochrane AH. Sero-epidemiological studies of malaria in indian tribes and monkeys of the Amazon basin of Brazil. Am J Trop Med Hyg. 1989;4:379–85.
Carréri-Bruno GC, Ciaravolo RM, Pereira M. Malaria acquired during entomological research in the Serra do Mar, southeastern region of Brazil. Rev Saude Publica. 1995;29:142–3.
Singh B, Kim Sung L, Matusop A, Radhakrishnan A, Shamsul SS, Cox-Singh J, et al. A large focus of naturally acquired Plasmodium knowlesi infections in human beings. Lancet. 2004;363:1017–24.
Ta TH, Hisam S, Lanza M, Jiram AI, Ismail N, Rubio JM. First case of a naturally acquired human infection with Plasmodium cynomolgi. Malar J. 2014;13:68.
Yusof R, Lau YL, Mahmud R, Fong MY, Jelip J, Ngian HU, et al. High proportion of knowlesi malaria in recent malaria cases in Malaysia. Malar J. 2014;13:168.
Coatney GR. The simian malarias: zoonoses, anthroponoses, or both? Am J Trop Med Hyg. 1971;20:795–803.
Deane LM. Simian malaria in Brazil. Mem Inst Oswaldo Cruz. 1992;87 Suppl 3:1–20.
Leclerc M, Hugot J, Durand P, Renaud F. Evolutionary relationships between 15 Plasmodium species from new and old world primates (including humans): an 18S rDNA cladistic analysis. Parasitology. 2004;129:677–84.
Duarte AM, Porto MA, Curado I, Malafronte RS, Hoffmann EH, de Oliveira SG, et al. Widespread occurrence of antibodies against circumsporozoite protein and against blood forms of Plasmodium vivax, P. falciparum and P. malariae in Brazilian wild monkeys. J Med Primatol. 2006;35:87–96.
Garnham PCC. Malaria parasites and other haemosporidia. Oxford: Blackwell; 1966.
Lourenço-de-Oliveira R, Deane LM. Simian malaria at two sites in the Brazilian Amazon. I – The infection rates of Plasmodium brasilianum in non-human primates. Mem Inst Oswaldo Cruz. 1995;90:331–9.
Deane LM, Ferreira Neto J, Sitônio JG. A new natural host of Plasmodium simium and Plasmodium brasilianum: the woolly spider monkey. Rev Inst Med Trop Sao Paulo. 1968;10:287–8.
Duarte AM, Malafronte RS, Cerutti C, Curado I, de Paiva BR, Maeda AY, et al. Natural Plasmodium infections in Brazilian wild monkeys: reservoirs for human infections? Acta Trop. 2008;107:179–85.
Pina-Costa A, Brasil P, Santi SM, Araujo MP, Suárez-Mutis MC, Santelli AC, et al. Malaria in Brazil: what happens outside the Amazonian endemic region. Mem Inst Oswaldo Cruz. 2014;109:618–33.
Deane LM. Malaria studies and control in Brazil. Am J Trop Med Hyg. 1988;38:223–30.
Oliveira-Ferreira J, Lacerda MV, Brasil P, Ladislau JL, Tauil PL, Daniel-Ribeiro CT. Malaria in Brazil: an overview. Malar J. 2010;9:115.
Miguel RB, Peiter PC, Albuquerque H, Coura JR, Moza PG, Costa Ade P, et al. Malaria in the state of Rio de Janeiro, Brazil, an Atlantic Forest area: an assessment using the health surveillance service. Mem Inst Oswaldo Cruz. 2014;109:634–40.
Yamasaki T, Summa ME, Neves DV, de Oliveira SG, Gomes AC. Natural Plasmodium infections in Brazilian wild monkeys: reservoirs for human infections? Acta Trop. 2008;107:179–85.
Snounou G, Viriyakosol S, Jarra W, Thaithong S, Brown KN. Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detection of a high prevalence of mixed infections. Mol Biochem Parasitol. 1993;58:283–92.
Tamura K. Estimation of the number of nucleotide substitutions when there are strong transition-transversion and G + C-content biases. Mol Biol Evol. 1992;9:678–87.
Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S. MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol. 2011;28:2731–9.
Lourenço-de-Oliveira R. Natural infection of golden lion tamarin, Leontopithecus rosalia, with Trypanosoma cruzi in the state of Rio de Janeiro. Mem Inst Oswaldo Cruz. 1990;85(I):15.
Moreira GV, Peixoto CMS, Ziccardi M, Oliveira RL, Castro MG, Dionísio DF, et al. Prevalência de Trypanosoma cruzi, Trypanosoma minasense e de anticorpos contra arbovírus em primatas não humanos (Callithrichidae) em cativeiro. Rev Bras Med Vet. 2000;22:252–4.
Fragaszy DM, Visalberghi E, Fedigan LM. The complete Capuchin – the biology of the genus Cebus. Cambridge: Cambridge University Press; 2004.
Izawa K. Foods and feeding behavior of wild black-capped capuchin (Cebus apella). Primates. 1979;20:57–76.
Freese CHE, Oppenheimer JR. The Capuchin Monkeys, Genus Cebus. In: A. & R. Mittermeier, editor. Ecology and behavior of neotropical primates. Volume 1. 1st ed. Rio de Janeiro: Academia Brasileira de Ciências; 1981. p. 331–90.
Brown A, Chalukian S, Malmierca L. Habitat y alimentacion de Cebus apella en el N.O. Argentino y la disponibilidad de frutos en el dosel arboreo. Revista del Museuo Argentino de Ciencias Naturales. 1984;13:273–80.
Fedigan LM. Vertebrate predation in Cebus capucinus: meat eating in a Neotropical monkey. Folia Primatol. 1990;54:196–205.
Alfaro JW, Silva Jr JD, Rylands AB. How different are robust and gracile capuchin monkeys? An argument for the use of Sapajus and Cebus. Am J Primatol. 2012;74:273–86.
Araújo MS, Messias MR, Figueiró MR, Gil LH, Probst CM, Vidal NM, et al. Natural Plasmodium infection in monkeys in the state of Rondônia (Brazilian Western Amazon). Malar J. 2013;12:180.
Deane LM. Studies on simian malaria in Brazil. Bull World Health Organ. 1964;31:752–3.
Flores-Mendoza C, Lourenço-de-Oliveira R. Bionomics of Anopheles aquasalis Curry 1932, in Guaraí, State of Rio de Janeiro, southeastern Brazil-I. Seasonal distribution and parity rates. Mem Inst Oswaldo Cruz. 1996;91:265–70.
Miguel RB. Estudo da infecção humana por Plasmodium spp no município de Guapimirim, estado do Rio de Janeiro. Fundação Oswaldo Cruz: MsD thesis. Instituto Oswaldo Cruz; 2011.
Curado I, Dos Santos MR, de Castro Duarte AM, Kirchgatter K, Branquinho MS, Bianchi Galati EA. Malaria epidemiology in low-endemicity areas of the Atlantic Forest in the Vale do Ribeira, São Paulo, Brazil. Acta Trop. 2006;100:54–62.
Tazi L, Ayala FJ. Unresolved direction of host transfer of Plasmodium vivax v. P. simium and P. malariae v. P. brasilianum. Infect Genet Evol. 2011;11:209–21.
Escalante AA, Barrio E, Ayala FJ. Evolutionary origin of human and primate malarias: evidence from the circumsporozoite protein gene. Mol Biol Evol. 1995;12:616–26.
Escalante AA, Cornejo OE, Freeland DE, Poe AC, Durrego E, Collins WE, et al. A monkey’s tale: the origin of Plasmodium vivax as a human malaria parasite. Proc Natl Acad Sci U S A. 2005;102:1980–5.
Mu J, Joy DA, Duan J, Huang Y, Carlton J, Walker J, et al. Host switch leads to emergence of Plasmodium vivax malaria in humans. Mol Biol Evol. 2005;22:1686–93.
Carter R. Speculations on the origins of Plasmodium vivax malaria. Trends Parasitol. 2003;19:214–9.
Lim C, Tazi L, Ayala F. Plasmodium vivax: recent world expansion and genetic identity to Plasmodium simium. Proc Natl Acad Sci U S A. 2005;102:15523–8.
Cornejo OE, Escalante AA. The origin and age of Plasmodium vivax. Trends Parasitol. 2006;22:558–63.
Li J, Collins WE, Wirtz RA, Rathore D, Lal A, McCutchan TF. Geographic subdivision of the range of the malaria parasite Plasmodium vivax. Emerg Infect Dis. 2001;7:35–42.
Taylor JE, Pacheco MA, Bacon DJ, Beg MA, Machado RL, Fairhurst RM, et al. The evolutionary history of Plasmodium vivax as inferred from mitochondrial genomes: parasite genetic diversity in the Americas. Mol Biol Evol. 2013;30:2050–64.
Pacheco MA, Cranfield M, Cameron K, Escalante AA. Malarial parasite diversity in chimpanzees: the value of comparative approaches to ascertain the evolution of Plasmodium falciparum antigens. Malar J. 2013;12:328.
World Health Organization (WHO). MALVAC meeting 2004: evaluation of malaria vaccines. Pre-clinical evaluation group: optimizing the developmental pathway from the lab to the clinic. Switzerland: Report from a technical consultation at WHO/IVR Malaria Vaccine Advisory Committee meeting Montreux; 2004.
The authors thank the PDTIS sequencing facilities of FIOCRUZ, Gabriela Assis and Lara Cotta Amaral for assistance on sequencing experiments and the team of the Primate Centre of Rio de Janeiro (CPRJ/INEA). The authors are also grateful to the Parc Zoologique et Botanique Ville de Mulhouse, France; to Jean Marc Lernould, Zoological Society for the Conservation of Species and Populations – München Germany; and to Dr Roland Wirth and the Conservation International (CI), for cooperation in the program for reproduction of Sapajus from the Brazilian Atlantic Forest. CFAB, CTDR and RLO are supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (through a Research Fellowship). CTDR and RLO are grateful for the support of the Faperj as Cientistas do nosso Estado. APC and DAMA received fellowships from the Coordenação de Pessoal de Ensino Superior (Capes). The study received financial support from the Secretary for Health Surveillance (SVS) through the Global Fund.
The authors declare that they have no competing interests.
CTDR, CFAB, PB, APC, MGZ, and RLO conceived the idea and participated in the study design. AP, PB, MCS, and APC were responsible for sample collection. DAMA and TNS carried out the experiments and analysed the data. CFAB, CTDR, RLO, and DAMA drafted the manuscript. All authors read, corrected and approved the final manuscript.
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Cite this article
de Alvarenga, D.A.M., de Pina-Costa, A., de Sousa, T.N. et al. Simian malaria in the Brazilian Atlantic forest: first description of natural infection of capuchin monkeys (Cebinae subfamily) by Plasmodium simium . Malar J 14, 81 (2015). https://doi.org/10.1186/s12936-015-0606-6
- Simian malaria
- Plasmodium simium
- Plasmodium vivax
- Plasmodium brasilianum
- Plasmodium malariae
- Molecular diagnosis
- Atlantic forest


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PITTMAN, JUSTICE, FOR THE COURT:
The Mississippi State Bar filed a Formal Complaint against Donald J. Steighner, alleging that he had violated several provisions of the Code of Professional Responsibility, which was in effect at that time, and Miss. Code Ann. 73-3-35 (1972). A trial on the merits was conducted by a Complaint Tribunal. The Tribunal found that Steighner had violated the provisions of DR 6-101 (A) (2 and 3) and DR 7-101 (A) (1 and 2) of the Code of Professional Responsibility and imposed a thirty (30) day suspension. Steighner appeals, assigning as error:
I. THE DECISION OF THE COMPLAINT TRIBUNAL FINDING THE APPELLANT IN VIOLATION OF THE CODE OF PROFESSIONAL CONDUCT WAS INCORRECT AND NOT SUPPORTED BY THE EVIDENCE.
In 1983 Phyllis Taggart was experiencing medical problems, and as a result consulted with Dr. Perrin Smith, a gynecologist. Dr. Smith performed a hysterectomy on June 9, 1983, in the Golden Triangle Medical Center in Columbus, Mississippi. Mrs. Taggart was discharged on June 17, 1983. She was re-admitted for surgery on June 25, 1983, and was operated on by Dr. Miles, a partner of Dr. Smith. Sometime after that, in June or July, 1983, Mrs. Taggart retained Mr. Steighner on a contingency-fee basis to represent her on a malpractice claim. Mr. Steighner told Mrs. Taggart that he would need a complete set of her medical records.
Mr. Steighner received Mrs. Taggart's medical records, although he could not say when. Mr. Steighner claimed that from the time he was retained until May, 1984, he was keeping in touch with Mrs. Taggart via telephone, checking on how her health was affecting her daily activities. Steighner reviewed Mrs. Taggart's medical records, and had his wife, who was an R.N., review them. Steighner spoke to a Dr. Sams, who was the anesthesiologist during Mrs. Taggart's surgery. Steighner reported that Sams felt that there was nothing unusual about the case. Steighner did not get a statement from Sams, nor did he depose him. Steighner also read the notes made by the nurses who were at the surgery. According to Steighner, the notes indicated nothing unusual, although he did not get statements from any of the nurses. Mr. Steighner was able to find out who Dr. Smith's insurance agent was, but little else. According to Steighner, Dr. Miles, who had assisted Dr. Smith during the June 9 surgery, and had performed the June 25 surgery, would not talk to him at all. As of May, 1984, Steighner had not found any expert witnesses who would testify that the doctors who had operated on Mrs. Taggart had been negligent or had deviated from accepted standards of practice.
On May 23, 1984, Steighner wrote to Mrs. Taggart, stating
that he had been reading her medical records and needed more clarification on the facts. He asked Mrs. Taggart to compile for him a complete summary of her medical problems leading up to and including the surgeries in question. Mrs. Taggart complied with this request in a timely fashion. On June 4, 1985, Steighner filed suit in U.S. District Court in the Northern District of Mississippi, with the complaint styled "Phyllis Taggart and Charles L. Taggart v. The Columbus Women's Clinic and Dr. Perrin Smith." The Columbus Women's Clinic was later dismissed from the suit, as Mrs. Taggart was never hospitalized there. The primary reason the suit was filed was to toll the running of the statute of limitations.
In June, 1985, Mrs. Taggart, who had moved with her family to East Moline, Illinois in January, 1984, came back to Columbus to be deposed. This was the only deposition that Steighner took. He also did not file any interrogatories and did not request production of any documents, saying that he already had "all the documents I needed - that existed." He allegedly consulted with at least two doctors. The first was a Phillipine doctor, but Steighner did not talk to him personally and could not remember his name. The Phillipine doctor did suggest to Steighner's associate that they consult with Dr. John Wilson, an emergency room physician in Tupelo who had a background in forensic medicine. According to Steighner, Wilson reviewed the file and concluded that while Dr. Smith's bedside manner could be improved, there was nothing wrong with the surgery that he had performed, and Mrs. Taggart was the victim of an "untoward incident" . Steighner claimed that Dr. Wilson dictated a statement of his opinion, but could not find the tape, and the taped statement had never been reduced to writing. Steighner never reduced the statement to writing because he did not want it to be discoverable. Steighner did not consult with Dr. Wilson until after December 20, 1985.
On December 20, 1985, Dr. Smith filed a Motion for Summary Judgment. Steighner never filed any response to this Motion. Steighner never wrote to Mrs. Taggart to inform her about the Motion. He did not remember whether he had sent a copy of the Motion to Mrs. Taggart. He was aware that a response was required within ten days. On February 25, 1986, Dr. Smith's Motion for Summary Judgment was granted by the Court. Steighner did not discuss appealing the decision with Mrs. Taggart. Although Steighner claimed to have told Mrs. Taggart that "she had no case" , he never informed her that if her suit was dismissed, she had no further recourse against Dr. Smith and Golden Triangle Medical Center. Steighner did not remember Mrs. Taggart ever requesting her file so that another attorney might review it. Steighner never wrote to Mrs. Taggart to inform her about his opinion that there was no case and that it would soon
be dismissed "[b]ecause we spoke so often together. I felt that we had that communication. She was forever calling me and I felt we had that communication." As to why he did not take a voluntary nonsuit against the defendants instead of allowing the summary judgment motion to ripen, Steighner claimed that he just kept looking and hoping that something would turn up to support the suit. Mrs. Taggart has not sued Steighner for malpractice.
According to Mrs. Taggart, she began experiencing problems after the June 9 hysterectomy, until she finally had to be re-admitted for surgery on June 25, 1983. This surgery was performed by Dr. Miles, Dr. Smith's partner. It lasted four and a half hours, and Mrs. Taggart claimed that after the emergency surgery, Dr. Miles "told my husband and I that he had to take out part of my bowels and clean them up that they had been nicked up by Dr. Smith and that my stomach lining had been nicked up." According to Mrs. Taggart, she was not charged anything for the second surgery because of this. Mrs. Taggart believed that she paid Steighner $100.00 so that he could obtain her medical records. Steighner also told Mrs. Taggart that once he got her medical records, he would send them to Jackson to be examined by a panel of doctors. She complained that she never could seem to contact Steighner at his office, and when she could he was not very forthcoming, so she began to call him at his home, usually around 6:00 A.M. After Mrs. Taggart was deposed, she claimed that Steighner told her that Dr. Smith would be deposed the next day. Mrs. Taggart asked that Steighner contact her and tell her how it went. When he did not contact her, she called to find out what had happened, and Steighner told her first, that he had been unable to set up the deposition, and later told her that Dr. Smith was too busy and "[t]hat he would just lie about it anyway."
The next contact Mrs. Taggart had with Steighner came after the granting of the summary judgment motion. When Mrs. Taggart asked Steighner why the suit had been dismissed, he told her that she would have to get a doctor to explain it. It was at this time that she first considered filing a complaint with the Bar. Mrs. Taggart also contacted two Mississippi attorneys, Mr. Carter and Mr. Liston, to advise her on her further options against the doctors or against Steighner. She decided not to sue Steighner. The only expenses Mrs. Taggart could remember incurring were for the first set of medical records that Steighner requested, and approximately $68.00 for a filing fee. Mrs. Taggart felt that Steighner had written her "maybe a couple" of times besides the May 23, 1984 letter. Mrs. Taggart estimated that between June, 1983, and February, 1986, she had telephoned Steighner two hundred times, although she could not remember the amounts of any of her bills. She remembered Steighner calling her a couple of times. He notified her of the dismissal of the suit by telephone. When Steighner sent Mrs. Taggart a copy of the
judgment of dismissal, he did not include any kind of cover letter or explanation.
The Mississippi Bar Association filed a Formal Complaint against Steighner on March 4, 1987. The Complaint alleged that Steighner had violated certain provisions of the Code of Professional Responsibility, including DR 1-102 (A)(4, 5, and 6), DR 6-101 (A)(2 and 3), DR 7-101 (A)(1 and 3), and Miss. Code Ann. 73-3-35 (1972). The Complaint asked the Tribunal to take appropriate disciplinary action against Steighner, and to order him to pay $38.17, the costs of filing the Formal Complaint, along with the costs and expenses of the litigation of the Formal Complaint. Chief Justice Roy Noble Lee designated John C. Love, Jr., John W. Prewitt, Sr., and Fred Ross, Jr. to sit as members of the Complaint Tribunal.
The Bar made a Motion for Entry of Default Judgment on May 27, 1987. In its Motion the Bar alleged that Steighner had been served with a copy of the Formal Complaint on April 10 and April 21, 1987, that he had not answered the Complaint within thirty days, and that a default judgment should be entered against Steighner. Steighner finally answered the Complaint on June 11, 1987, denying any deficient conduct. Steighner also moved to set aside the Bar's Default Motion, alleging that he had valid defenses to the Complaint and had had a heavy caseload during this time. A hearing was held by the Complaint Tribunal on the Default Motion on June 16, 1987. Because of Steighner being a solo practitioner who had been involved in several felony criminal cases, and ...

domain: law


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RECORD BREAKING YEAR OF HOLIDAY ACTIVITY PROVISION
This can result in ‘unhealthy holidays’ - increased holiday hunger, limited access to nutritious and healthy meals alongside limited access to physical and fun activities. All of which can contribute to the likelihood of experiencing social isolation. Sadly, these issues have also been further exasperated by the Covid-19 pandemic.
In response to this, the Department of Education (DfE) has provided local authorities with Holiday Activity Funding (HAF) to develop a free activity and food programme during the holidays. This means we are able to work with clubs and organisations across the city to develop a food and activity programme to respond to the above, which we know will make a positive impact on the children and young people of Manchester.
From Summer 2021 - Summer 2022, MCRactive and Manchester City Council offered thousands of FREE places to children eligible for benefit related free school meals and those with no recourse to public funds; to access a variety of activities over the school holiday periods. Activities range from Sport and Physical Activities, Arts and Crafts and sometimes can include a variety of Day Trips.
Our work with holiday activity has gone from strength to strength over the past year, with a focus on providing support for children and families eligible for free school meals, with funding secured from the Department for Education; allowing us to deliver enriching and inclusive activities alongside a healthy meal.
This year, Manchester commissioned 115 unique providers to receive Holiday Activity Funding (HAF) to deliver a meal and activities over school holidays. Aimed at local 5-16 years old in receipt of free school meals, we delivered;
- 3,466 Sessions
- 81,847 engagements
- 14,572 children
MCRactive, the City Council are immensely proud of the work we have achieved over the past year, but we couldn't have done it without the hard work or local providers, volunteers and communities working tirelessly to provide free, engaging and vital activity and hot meals to disadvantaged children in our city. And we have no plans to stop anytime soon, with plans to expand the Holiday Activity Fund program even further in 2023 to ensure we reach all children in need accross Manchester.

domain: news


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THE 10 BEST Chinese Restaurants in Hamilton, Updated April - Tripadvisor
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domain: food


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In this outing Caesar and his growing family have made lives for themselves in the forest. The renegade gorilla Kubo, from the second film who wanted to destroy humanity, is long gone. All is well until humans bring war to the group of apes who just want to survive and prosper in peace. Caesar walks a deadly line where he will either become like Kubo to save his family or face extinction. That moral dilemma of either becoming like your enemy or transcending to a higher place is part of what makes “Apes” tick.
“War for the Planet of the Apes” isn’t a perfect film but the flaws are few and in between. “Apes” once again stars the very impressive Andy Serkis as Caesar and a bunch of green dots that enable him to appear as an erect walking and talking chimpanzee. Weta Digital who helped develop the special effects has truly outdone themselves. Of course people generally don’t go to a film just to see the special effects but this film could make a case for exactly that. Skillfully, it doesn’t have too because the film delivers on much more than just how realistic the actors look as apes.
Adding to the film’s positive landscape is another bold and dynamic soundtrack from Michael Giachina (Rogue One, Doctor Strange) who is innately aware that a large portion of the actors are using sign language to communicate. His music takes great care to evoke emotions alongside actors who don’t get to vocalize a lot of audible language. Giachina even includes a few riffs of the original “Planet of the Apes” theme if you listen for it.
Reeves and his crew do an excellent job of creating an atmosphere of oppressiveness where the apes live in a world of rainy, cloudy environments. The color palettes are dreary and are expertly designed around characters who won’t be vocalizing. The environment and staging of the scenes go a long ways in helping to tell this story and it’s an easy bet that some nominations will be coming down later this year for the design work done on this film (besides the technical aspects which will be easily nominated as well).
All three films combined can go up against any re-imagined and or rebooted franchise films in the past twenty years and be cited as the best. That’s something to be said but when you really get down to comparing it to franchise films that rely heavily on action and go weak on character development and or story, “Apes” never forgets character development. Explosions, dramatic deaths, visceral fight scenes mean nothing if you don’t care about the characters. Reeves keeps that tightly in focus as he weaves a tale about survival of the fittest. As spectacular as the special effects are in this film, they wouldn’t mean anything if the actors didn’t have a meaningful story to perform. “War for the Planet of the Apes” is easily the best of the three in an already stellar franchise.
Final Verdict: 4 out of 5

domain: artistic


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About Satanic Warmaster
Satanic Warmaster is a black metal band from Lappeenranta, Finland. The musician "Satanic Tyrant Werwolf" began recording under this name in 1999. Satanic Warmaster plays a raw style of black metal. The band's lyrics deal mainly with nature, folklore, Satanism, and "occult and warlike themes".


The Japanese carmaker chose to follow a different path from its rivals by its use of boxer engines and all-wheel drive across all models. The company is renowned and respected for its innovation, technology, and participation in motor sport. Subaru offers the small Impreza hatch and sedan, medium Liberty sedan, the high riding Liberty-based Outback crossover wagon, XV and Forester SUVs, BRZ sports coupe, and rally-bred WRX and WRX STi hatches.
Latest Subaru Reviews
Family SUV Comparison: Haval H6 Hybrid Ultra vs Mazda CX-5 Touring Active 2.2D AWD vs Subaru Forester Hybrid L vs Toyota RAV4 GXL Hybrid16 Jul 2022 by Richard Berry
Kia Sportage GT-Line vs Mitsubishi Outlander Exceed vs Subaru Forester 2.5i-S vs Hyundai Tucson Highlander N Line 2022 comparison review16 Jan 2022 by Richard Berry
Ford Puma and Subaru XV review - We compare two of Australia's most distinctive small SUVs7 Oct 2021 by Tom White
Latest Subaru News
Exxy Rexy! 2023 Subaru WRX pricing goes up with no updates or extra features for Volkswagen Golf GTI, Hyundai i30 N rival - but by how much?
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No car is perfect, so we've gathered everything relating to the Subaru reliability here to help you decide if it's a smart buy.


We gather weekly the most important Vaping news.
South Africa flirts with e-cigarette tax which could set back the fight against smoking
The South African government intends to introduce a new excise tax on e-cigarettes, applying the duty to both non-nicotine and nicotine solutions used in these devices. Harm reduction advocates have expressed their alarm over the proposed rules, which could see e-cigarette products regulated and taxed in a similar vein to combustible tobacco products.
Denmark proposes ban on selling cigarettes to people born after 2010
Denmark has unveiled plans to ban the sale of cigarettes and nicotine products to any citizens born after 2010. The move aims to prevent the next generation of Danes from touching any form of tobacco, Health Minister Magnus Heunicke told a press conference.
Smoke-free products to help Philippines reduce healthcare expenditures, say experts
Making e-cigarettes and heated tobacco products available as less harmful alternatives to traditional cigarettes for millions of Filipino smokers will enable the government to reduce the smoking rate and manage healthcare expenditures, international public health policy experts said.
‘Time is running out’: Connecticut advocates push for ban on flavored vaping products
Despite tighter federal restrictions on flavored e-cigarette products that took effect in 2020, supporters of a Connecticut bill say the state needs to implement its own ban on the sale of these products in order to drive down use among youth.
Smoking spoils dental whiteness: the ODON study
In terms of public health, these studies may have a tremendous impact. People who feel that bad breath or their teeth appearance are a major problem, can be massively influenced by aesthetic considerations to stop smoking.
The Impact of Banning Electronic Nicotine Delivery Systems on Combustible Cigarette Sales: Evidence From US State-Level Policies
In the fall of 2019, several US states passed short-term bans on the sale of electronic nicotine delivery systems (ENDS) in response to an outbreak of illnesses strongly linked to tetrahydrocannabinol vaping products that received national news coverage. This study assessed how such state-level ENDS bans in 3 US states may have affected cigarette sales.
Nicotine use and stress
Using nicotine can be complicated. In the short-term, vaping nicotine may feel good by stimulating production of the “feel good” chemical in the brain called dopamine which can create feelings of pleasure and relaxation. Vaping nicotine also creates social opportunities to connect with other people and provides a distraction from stressful situations


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Bridgton, Maine Treatment Centers
Bridgton, ME 04009 View Details →
Giving the Alcoholic Person a Successful Start on the Twelve-Step Road of Recovery Since 1977 The New Direction Alcohol Rehabilitation in New York The New Direction was founded in 1977 by Matt and Margaret Batson under the premise that people who make a commitment to A.A. tend to stay sober. Since the doors of the….
It’s hard to say 30 strikes and you’re out with a straight face. It sounds like a silly joke. But the astounding truth is that the Vancouver Police Department tracks 379 offenders who have an average of 39 convictions each, primarily for theft and other property crime. Six have more than 100 convictions. These are….
Regardless of the type of drug involved, addictive drugs all exert certain effects that increase the likelihood of ongoing drug use. Also known as psychoactive agents, these drugs are able to cross the blood-brain barrier, which gives them easy access to the brain’s workings. According to the National Institute on Drug Abuse, drug addiction creates….
It’s a success story in an area with too few success stories. When she was 19 years old, Kim Selby came to Humboldt County to grow pot. Then she became addicted to methamphetamine. Today, she is off the drug and runs a clean and sober house for women in Fortuna. The nine women who live….
Anyone who’s battled a heroin addiction well knows how hard this drug can be on the body. While the “highs” from heroin may feel great, the lows can be equally painful. According to the National Institute on Drug Abuse, in 2011, as many as 4.2 million Americans reported having used heroin on at least one….
Where do calls go?
Calls to numbers on a specific treatment center listing will be routed to that treatment center. Calls to any general helpline (non-facility specific 1-8XX numbers) could be forwarded to SAMHSA or a verified treatment provider. Calls are routed based on availability and geographic location.

domain: medical


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Essentia Health Joins the Cardiometabolic Center Alliance as a Charter Member
April 22, 2022 | Press Releases
The Cardiometabolic Center Alliance (CMCA), founded by Saint Luke’s Mid America Heart Institute, is honored to announce the addition of Essentia Health as a Charter Member.
As a coalition that seeks to establish Cardiometabolic Centers of Excellence nationwide aimed at transforming the treatment of type 2 diabetes (T2D), and related cardiovascular (CV) and renal comorbidities, the CMCA considers Essentia Health—an integrated health system spanning the upper Midwest—a key member of the Alliance. Essentia Health will house their program in Duluth and Fargo cardiology practices.
“We’re thrilled to join the Cardiometabolic Center Alliance, a nationwide initiative designed to improve care and outcomes for patients with cardiometabolic diseases, such as type 2 diabetes and elevated cholesterol,” said Dr. Natalia Rocha, a physician championing Essentia’s program. “In our cardiometabolic clinic, we will tailor a personalized, multidisciplinary plan of care following up to date guidelines to ensure risk factors for heart disease are well controlled.”
“The inclusion of Essentia Health as a member of the CMCA speaks to the mission of the Alliance which is to transform the quality of care delivered to those living with cardiometabolic disease,” said Mikhail Kosiborod, MD, Executive Director of the CMCA. “Essentia’s geographic reach ensures that team-based, coordinated care is delivered to those in need—regardless of where they live. We look forward to our collaboration with Essentia as we work together to improve outcomes for our patients.”
Dr. Catherine Benziger, another cardiologist leading Essentia’s center, added, “We are excited to join the Alliance because we believe it will help us deliver the very best care to patients. High-risk patients, including those with strong family history of heart problems, obesity, diabetes, high cholesterol, and prior heart disease, can get comprehensive care in the new cardiometabolic clinic. Our goal is to help individuals live a more heart-healthy life free of heart disease by losing weight, getting their LDL cholesterol and hemoglobin A1c under control, and overall feeling better.”
About the Cardiometabolic Center Alliance
The Cardiometabolic Center Alliance is a national collaborative of healthcare organizations committed to the mission of improving the quality of care and outcomes in patients with cardiometabolic disease. Across its aligned member sites, care is delivered via a standardized, patient-centered, team-based, comprehensive risk-reduction approach based on a novel care model designed and successfully implemented by Saint Luke’s Michael & Marlys Haverty Cardiometabolic Center of Excellence in Kansas City, Missouri.
About Essentia Health
Essentia Health is an integrated health system serving patients in Minnesota, Wisconsin, and North Dakota. Headquartered in Duluth, Minnesota, Essentia Health combines the strengths and talents of more than 14,000 employees, including more than 2,100 physicians and advanced practitioners, who serve our patients and communities through the mission of being called to make a healthy difference in people’s lives. Essentia Health, which includes many Catholic facilities, is guided by the values of Quality, Hospitality, Respect, Joy, Justice, Stewardship and Teamwork. The organization lives out its mission by having a patient-centered focus at 14 hospitals, 71 clinics, six long-term care facilities, three assisted living facilities, three independent living facilities, six ambulance services and one research institute. Essentia Health is accredited as an Accountable Care Organization by the National Committee for Quality Assurance.


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Flying the flag for UK IT The UK IT industry has dropped from third to sixth in a league table of IT industry competitiveness. Despite this, the annual research by the Economist Intelligence Unit...
Bitdefender's decryption tool can neutralise the latest versions of GandCrab, as well as retrieve files encrypted by earlier versions
Zhengfei admits he underestimated potential impact of being placed on US 'Entity List'
Huawei is considering pulling launch of Honor 20 smartphone from Europe in order to focus on China
President Trump labelled the New York Times report "a virtual act of treason"