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1.
Arch Virol ; 166(1): 275-280, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33201342

RESUMO

The infectivity of shrew-borne hantaviruses to humans is still unclear because of the lack of a serodiagnosis method for these viruses. In this study, we prepared recombinant nucleocapsid (rN) proteins of Seewis orthohantavirus, Altai orthohantavirus (ALTV), Thottapalayam thottimvirus (TPMV), and Asama orthohantavirus. Using monospecific rabbit sera, no antigenic cross-reactivity was observed. In a serosurvey of 104 samples from renal patients and 271 samples from heathy controls from Sri Lanka, one patient serum and two healthy control sera reacted with rN proteins of ALTV and TPMV, respectively. The novel assays should be applied to investigate potential infectivity of shrew-borne hantaviruses to humans.


Assuntos
Infecções por Hantavirus/imunologia , Infecções por Hantavirus/virologia , Orthohantavírus/imunologia , Musaranhos/virologia , Animais , Estudos de Casos e Controles , Linhagem Celular , Chlorocebus aethiops , Células HEK293 , Humanos , Proteínas do Nucleocapsídeo/imunologia , Filogenia , Vírus de RNA/imunologia , Coelhos , Proteínas Recombinantes/imunologia , Testes Sorológicos/métodos , Sri Lanka , Células Vero
2.
Virol J ; 11: 87, 2014 May 14.
Artigo em Inglês | MEDLINE | ID: mdl-24885901

RESUMO

BACKGROUND: Hantaviruses are causative agents of hemorrhagic fever with renal syndrome (HFRS) and nephropathia epidemica (NE) in the Old World and hantavirus pulmonary syndrome (HPS) in the New World. There is a need for time-saving diagnostic methods. In the present study, recombinant N antigens were used as antigens in an immunochromatography strip (ICG) test to detect specific IgG antibodies. METHODS: The N-terminal 103 amino acids (aa) of Hantaan virus (HTNV), Puumala virus (PUUV) and Andes virus (ANDV) nucleocapsid (N) protein were expressed in E. coli as representative antigens of three groups (HFRS, NE and HPS-causing viruses) of hantavirus. Five different types of ICG test strips, one antigen line on one strip for each of the three selected hantaviruses (HTNV, PUUV and ANDV), three antigen lines on one strip and a mixed antigen line on one strip, were developed and sensitivities were compared. RESULTS: A total of 87 convalescent-phase patient sera, including sera from 35 HFRS patients, 36 NE patients and 16 HPS patients, and 25 sera from healthy seronegative people as negative controls were used to evaluate the ICG test. Sensitivities of the three-line strip and mixed-line strip were similar to those of the single antigen strip (97.2 to 100%). On the other hand, all of the ICG test strips showed high specificities to healthy donors. CONCLUSION: These results indicated that the ICG test with the three representative antigens is an effective serodiagnostic tool for screening and typing of hantavirus infection in humans.


Assuntos
Anticorpos Antivirais/sangue , Cromatografia de Afinidade/métodos , Vírus Hantaan/imunologia , Infecções por Hantavirus/diagnóstico , Proteínas do Nucleocapsídeo , Orthohantavírus/imunologia , Virus Puumala/imunologia , Antígenos Virais/genética , Antígenos Virais/isolamento & purificação , Escherichia coli/genética , Expressão Gênica , Vírus Hantaan/genética , Orthohantavírus/genética , Infecções por Hantavirus/virologia , Humanos , Imunoglobulina G/sangue , Proteínas do Nucleocapsídeo/genética , Proteínas do Nucleocapsídeo/isolamento & purificação , Virus Puumala/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Sensibilidade e Especificidade
3.
Artigo em Inglês | MEDLINE | ID: mdl-23077817

RESUMO

To help formulate a local intervention for leptospirosis in Sri Lanka, we determined the serogroups of leptospiral species among 97 patients diagnosed with leptospirosis at the University of Peradeniya Teaching Hospital, Sri Lanka. Ninety-two point eight percent of the patients were men; nearly two-thirds were > or = 35 years old; the majority had secondary or higher education level, half were farmers or laborers; and 57.7% presented in the acute-phase of the illness. Twenty-five patients (25.8%) were confirmed to have leptospirosis by a positive laboratory method; 17 and 8 cases were confirmed with a positive test by quantitative MAT and nested PCR, respectively. Of the 17 MAT positive cases, infection occurred in a variety of serogroups, but the predominant groups were Sejroe and Tarassovi. Of the 8 nested PCR positive cases, 7 were seen among those with a MAT titer <200 and 1 occurred in a patient with a MAT titer > or = 200 but <400. Of the 8 PCR positive cases, 7 were infected with the leptospiral species L. interrogans. Approximately 26% of the clinically diagnosed patients were confirmed by the two laboratory methods. Laboratory positivity was based on the time of blood collection after the onset of fever. Further studies are warranted to refine the clinical diagnostic criteria and to develop more efficient and accurate diagnostic tests for leptospirosis in resource limited settings.


Assuntos
Infecção Hospitalar/epidemiologia , Leptospira/genética , Leptospirose/epidemiologia , Adulto , Sequência de Bases , Infecção Hospitalar/diagnóstico , Feminino , Genes Bacterianos , Hospitais Universitários , Humanos , Leptospirose/diagnóstico , Masculino , Dados de Sequência Molecular , Análise de Sequência de DNA , Fatores Socioeconômicos , Sri Lanka/epidemiologia
4.
J Am Vet Med Assoc ; 260(13): 1589-1596, 2022 07 25.
Artigo em Inglês | MEDLINE | ID: mdl-35895801

RESUMO

Leptospirosis is a quintessential one health disease of humans and animals caused by pathogenic spirochetes of the genus Leptospira. Intra- and interspecies transmission is dependent on 1) reservoir host animals in which organisms replicate and are shed in urine over long periods of time, 2) the persistence of spirochetes in the environment, and 3) subsequent human-animal-environmental interactions. The combination of increased flooding events due to climate change, changes in human-animal-environmental interactions as a result of the pandemic that favor a rise in the incidence of leptospirosis, and under-recognition of leptospirosis because of nonspecific clinical signs and severe signs that resemble COVID-19 represents a "perfect storm" for resurgence of leptospirosis in people and domestic animals. Although often considered a disease that occurs in warm, humid climates with high annual rainfall, pathogenic Leptospira spp have recently been associated with disease in animals and humans that reside in semiarid regions like the southwestern US and have impacted humans that have a wide spectrum of socioeconomic backgrounds. Therefore, it is critical that physicians, veterinarians, and public health experts maintain a high index of suspicion for the disease regardless of geographic and socioeconomic circumstances and work together to understand outbreaks and implement appropriate control measures. Over the last decade, major strides have been made in our understanding of the disease because of improvements in diagnostic tests, molecular epidemiologic tools, educational efforts on preventive measures, and vaccines. These novel approaches are highlighted in the companion Currents in One Health by Sykes et al, AJVR, September 2022.


Assuntos
COVID-19 , Leptospira , Leptospirose , Saúde Única , Humanos , Animais , COVID-19/veterinária , Leptospirose/veterinária , Surtos de Doenças , Zoonoses/epidemiologia
5.
Am J Vet Res ; 83(10)2022 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-35986911

RESUMO

Leptospirosis is an archetypal One Health problem as described in the companion Currents in One Health article in the October 2022 issue of the Journal of the American Veterinary Medical Association by Sykes et al. A thorough understanding of leptospirosis requires a detailed analysis of the elaborate interplay among pathogenic leptospiral strains, host species, and the environment. Such an understanding is required to inform appropriate preventative measures including vaccine design, prophylaxis efforts, educational programs that help to reduce exposure to pathogenic spirochetes, as well as policy development. Because of the complex epidemiology of leptospirosis, a One Health approach as defined by the One Health Initiative Task Force is critical-an approach that calls for "the collaborative efforts of multiple disciplines working locally, nationally, and globally, to attain optimal health for people, animals and our environment." Over the last three decades, progressive advances in cutting-edge molecular typing techniques, as well as our ability to rapidly generate and share large amounts of sequence data through establishment and growth of databases, have been central to accelerating a One Health understanding of the epidemiology of leptospirosis. Nevertheless, our dependence on serotype information because of the serovar-specific nature of current vaccines means that laborious serotyping efforts continue. With the advent of new approaches such as mRNA vaccines that are based on lipopolysaccharide immunogens, sequence- and/or proteomics-based typing methods may replace these methods.


Assuntos
Leptospira , Leptospirose , Saúde Única , Animais , Humanos , Leptospira/genética , Leptospirose/epidemiologia , Leptospirose/prevenção & controle , Leptospirose/veterinária , Tipagem Molecular/veterinária , Sorogrupo
6.
J Med Microbiol ; 71(12)2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36748416

RESUMO

Background. Chronic kidney disease of unknown aetiology (CKDu) is a major public health problem in Sri Lanka, especially among agrarian communities. Although the cause of CKDu is still unknown, hantavirus infection has been proposed as a risk factor.Methods. This study was performed using serological samples collected from two CKDu-endemic areas, Anuradhapura (2010) and Badulla districts (2010 and 2016), and a non-endemic area, Matale (2016) district. The presence of anti-Thailand orthohantavirus IgG antibodies was investigated in serum samples. Hantavirus seroprevalence and demographic data were epidemiologically analysed.Results. Seroprevalence was higher in CKDu patients (40.6-60.0 %) and healthy individuals in CKDu-endemic areas (17.6-25.5 %) than in healthy individuals in non-endemic areas (3.0 %). Statistically significant odds ratios (ORs) for hantavirus infection in CKDu patients were detected in CKDu-endemic areas [ORs: 3.2 and 3.1; 95 % confidence interval (CI): 1.8-5.5 and 1.8-5.2 in Anuradhapura and Badulla districts in 2010; and OR: 4.4, 95 % CI: 2.3-8.5 in 2016 in Badulla district). Furthermore, the OR for hantavirus infection in Badulla district has increased in the last decade from 3.1 (95 % CI: 1.8-5.3) to 4.4 (95 % CI: 2.3-8.5).Conclusion. Hantavirus infection has been prevalent in two distant CKDu-endemic areas since 2010. The observed significant association of hantavirus seropositivity with CKDu indicates a possible role of hantavirus infection in CKDu pathogenesis.


Assuntos
Infecções por Hantavirus , Insuficiência Renal Crônica , Humanos , Doenças Renais Crônicas Idiopáticas , Estudos Retrospectivos , Sri Lanka/epidemiologia , Prevalência , Estudos Soroepidemiológicos , Fatores de Risco , Infecções por Hantavirus/complicações , Infecções por Hantavirus/epidemiologia , Insuficiência Renal Crônica/complicações , Insuficiência Renal Crônica/epidemiologia
7.
Viruses ; 13(10)2021 10 02.
Artigo em Inglês | MEDLINE | ID: mdl-34696414

RESUMO

We reported the genetic evidence of circulating hantaviruses from small mammals captured in a chronic kidney disease of unknown etiology (CKDu) hotspot area of Sri Lanka. The high seroprevalence of anti-hantavirus antibodies against Thailand orthohantavirus (THAIV) has been reported among CKDu patients and rodents in Sri Lankan CKDu hotspots. We captured 116 small mammals from CKDu endemic regions in the Polonnaruwa District of Sri Lanka. Seven animals (five out of 11 Mus booduga and two out of 99 Rattus rattus) were PCR-positive for the hantavirus. A rat-borne sequence was grouped with a THAIV-like Anjozorobe virus. In contrast, Mus-borne sequences belonged to the THAIV lineage, suggesting a novel orthohantavirus species according to the phylogenetic analyses and whole-genome comparisons. Our genetic evidence indicates the presence of two THAIV-related viruses circulating in this CKDu endemic area, suggesting a basis for further investigations to identify the infectious virus in patients with CKDu and the CKDu induction mechanism of these viruses.


Assuntos
Infecções por Hantavirus/epidemiologia , Orthohepadnavirus/isolamento & purificação , Animais , Doenças Endêmicas , Orthohantavírus/genética , Camundongos , Orthohepadnavirus/patogenicidade , Filogenia , Ratos , Insuficiência Renal Crônica/epidemiologia , Insuficiência Renal Crônica/etiologia , Roedores/virologia , Estudos Soroepidemiológicos , Sri Lanka/epidemiologia
8.
PLoS Negl Trop Dis ; 14(7): e0008437, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32701971

RESUMO

BACKGROUND: Leptospirosis is one of the most significant zoonoses across the world not only because of its impact on human and animal health but also because of the economic and social impact on agrarian communities. Leptospirosis is endemic in Sri Lanka where paddy farming activities, the use of draught animals in agriculture, and peridomestic animals in urban and rural areas play important roles in maintaining the infection cycle of pathogenic Leptospira, especially concerning animals as a potential reservoir. In this study, an environmental DNA (eDNA) metabarcoding methodology was applied in two different agro-ecological regions of Sri Lanka to understand the eco-epidemiology of leptospirosis. METHODOLOGY/PRINCIPAL FINDINGS: Irrigation water samples were collected in Kandy District (wet zone mid-country region 2) and Girandurukotte, Badulla District (intermediate zone low-country region 2); and analysed for the presence of pathogenic Leptospira, associated microbiome and the potential reservoir animals. Briefly, we generated PCR products for high-throughput sequencing of multiple amplicons through next-generation sequencing. The analysis of eDNA showed different environmental microbiomes in both regions and a higher diversity of Leptospira species circulating in Kandy than in Girandurukotte. Moreover, the number of sequence reads of pathogenic Leptospira species associated with clinical cases such as L. interrogans was higher in Kandy than in Girandurukotte. Kandy also showed more animal species associated with pathogenic bacterial species than Girandurukotte. Finally, several pathogenic bacterial species including Arcobacter cryaerophilus, responsible for abortion in animals, was shown to be associated with pathogenic Leptospira. CONCLUSIONS/SIGNIFICANCE: Leptospirosis has been considered to be endemic in wet regions, consistently, leptospiral sequences were detected strongly in Kandy. The great Leptospira species diversity in Kandy observed in this study shows that the etiological agents of leptospirosis in Sri Lanka might be underestimated. Furthermore, our eDNA metabarcoding can be used to discriminate bacterial and animal species diversity in different regions and to explore environmental microbiomes to identify other associated bacterial pathogens in the environment.


Assuntos
Água Doce/microbiologia , Leptospira/isolamento & purificação , Leptospirose/epidemiologia , Leptospirose/microbiologia , Irrigação Agrícola , Animais , DNA Bacteriano/genética , DNA Ambiental/genética , Humanos , Leptospira/classificação , Leptospira/genética , Filogenia , Sri Lanka/epidemiologia , Zoonoses/microbiologia , Zoonoses/transmissão
9.
PLoS Negl Trop Dis ; 14(8): e0008573, 2020 08.
Artigo em Inglês | MEDLINE | ID: mdl-32841248

RESUMO

BACKGROUND: Leptospirosis has gained much attention in Sri Lanka since its large outbreak in 2008. However, most of the cases were clinically diagnosed and information on Leptospira genotypes and serotypes currently prevailing in the country is lacking. METHODOLOGY/PRINCIPAL FINDINGS: We retrospectively analyzed 24 Leptospira strains from human patients as well as isolated and characterized three Leptospira strains from black rats using the microscopic agglutination test with antisera for 19 serovars and multilocus sequence typing. The isolates were identified as Leptospira borgpetersenii sequence types (STs) 143 and 144; L. interrogans STs 30, 34, 43, 44, 74, 75, 80, 308, 313, 314, 316, and 317; and L. kirschneri ST318. Six of the 15 STs were identified for the first time in this study. Five serogroups such as Autumnalis, Grippotyphosa, Hebdomadis, Javanica, and Pyrogenes were detected among the isolates. Contrary to previous studies, various genotypes including novel STs were isolated during an outbreak in Southern Province. L. borgpetersenii serogroup Javanica ST143 was isolated both from a human and black rat. CONCLUSIONS/SIGNIFICANCE: This study revealed that genetically diverse Leptospira strains currently circulate in Sri Lanka: some genotypes have been circulating and others have emerged recently, which may explain the recent surge of leptospirosis patients with varying clinical manifestations and frequent outbreaks of leptospirosis. Black rats were identified as the source of infection for humans, but reservoir animals for other genotypes remain unknown.


Assuntos
Genótipo , Leptospira/classificação , Leptospira/genética , Leptospira/isolamento & purificação , Leptospirose/microbiologia , Tipagem de Sequências Multilocus/métodos , Adolescente , Adulto , Idoso , Testes de Aglutinação , Animais , Criança , DNA Bacteriano/análise , Reservatórios de Doenças , Feminino , Humanos , Leptospirose/epidemiologia , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Análise de Sequência de DNA , Sorogrupo , Sorotipagem , Sri Lanka/epidemiologia , Adulto Jovem
10.
Artigo em Inglês | MEDLINE | ID: mdl-32974220

RESUMO

Murid and cricetid rodents were previously believed to be the principal reservoir hosts of hantaviruses. Recently, however, multiple newfound hantaviruses have been discovered in shrews, moles, and bats, suggesting a complex evolutionary history. Little is known about the genetic diversity and geographic distribution of the prototype shrew-borne hantavirus, Thottapalayam thottimvirus (TPMV), carried by the Asian house shrew (Suncus murinus), which is widespread in Asia, Africa, and the Middle East. Comparison of TPMV genomic sequences from two Asian house shrews captured in Myanmar and Pakistan with TPMV strains in GenBank revealed that the Myanmar TPMV strain (H2763) was closely related to the prototype TPMV strain (VRC66412) from India. In the L-segment tree, on the other hand, the Pakistan TPMV strain (PK3629) appeared to be the most divergent, followed by TPMV strains from Nepal, then the Indian-Myanmar strains, and finally TPMV strains from China. The Myanmar strain of TPMV showed sequence similarity of 79.3-96.1% at the nucleotide level, but the deduced amino acid sequences showed a high degree of conservation of more than 94% with TPMV strains from Nepal, India, Pakistan, and China. Cophylogenetic analysis of host cytochrome b and TPMV strains suggested that the Pakistan TPMV strain was mismatched. Phylogenetic trees, based on host cytochrome b and cytochrome c oxidase subunit I genes of mitochondrial DNA, and on host recombination activating gene 1 of nuclear DNA, suggested that the Asian house shrew and Asian highland shrew (Suncus montanus) comprised a species complex. Overall, the geographic-specific clustering of TPMV strains in Asian countries suggested local host-specific adaptation. Additional in-depth studies are warranted to ascertain if TPMV originated in Asian house shrews on the Indian subcontinent.


Assuntos
Variação Genética , Musaranhos , África , Animais , China , Índia , Nepal , Paquistão , Filogenia , Filogeografia
12.
Vector Borne Zoonotic Dis ; 19(11): 859-866, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31339833

RESUMO

We have reported high seroprevalence to Thailand orthohantavirus (THAIV) or THAIV-related orthohantavirus (TRHV) among patients with chronic kidney disease of unknown etiology in Girandurukotte, Sri Lanka. THAIV or TRHV infection is considered to be transmitted by rodent hosts in this area, but its reservoir rodents have not yet been identified. Hence, 116 rodents were captured, and seroprevalences were examined by indirect immunofluorescent antibody assay (immunofluorescence assay [IFA]) using antigens of THAIV strain Thai749-infected Vero E6 cells and recombinant nucleocapsid protein of THAIV expressed in Vero E6 cell. Molecular biological species identification of rodents was carried out by sequencing rag1, irbp, and mitochondrial cytb genes. The majority (112/116) of the captured rodents were lineage Ib of black rats (Rattus rattus). Among them, 19.6% (22/112) of the rats possessed antibodies against THAIV. Also, a lesser bandicoot rat (Bandicota bengalensis), which belongs to the Sri Lankan endemic genetic lineage, was seropositive (1/1). Two Mus booduga and one Murinae sp. were seronegative. Rodent sera showed less cross-reactivities to antigens of Vero E6 cells infected with Hantaan orthohantavirus (HTNV), Seoul orthohantavirus (SEOV), and Puumala orthohantavirus (PUUV) in IFA. These results suggest that the hantavirus present in rodents in Sri Lanka is related to THAIV or TRHV rather than to SEOV, HTNV, or PUUV. However, it might be serologically distinct from the prototype THAIV strain, Thai749, used in this study. This study revealed that black rats and lesser bandicoot rats belonging to Sri Lankan endemic lineages are possible reservoirs for THAIV or TRHV in Girandurukotte. Further multiple geographical studies are needed to confirm the THAIV or TRHV reservoir status of black and lesser bandicoot rats in Sri Lanka.


Assuntos
Reservatórios de Doenças/virologia , Infecções por Hantavirus/veterinária , Orthohantavírus/isolamento & purificação , Doenças dos Roedores/virologia , Animais , Técnica Indireta de Fluorescência para Anticorpo , Orthohantavírus/classificação , Orthohantavírus/imunologia , Infecções por Hantavirus/epidemiologia , Murinae/sangue , Murinae/virologia , Ratos , Insuficiência Renal Crônica , Doenças dos Roedores/epidemiologia , Roedores/classificação , Estudos Soroepidemiológicos , Sri Lanka/epidemiologia
13.
Viruses ; 11(8)2019 07 31.
Artigo em Inglês | MEDLINE | ID: mdl-31370348

RESUMO

Chronic kidney disease of unknown etiology (CKDu) imposes a substantial burden on public health in Sri Lankan agricultural communities. High seroprevalences of hantavirus have been reported in CKDu patients in several locations of Sri Lanka. We carried out a cross-sectional study followed by an unmatched case-control comparison in two geographically distinct areas of Sri Lanka, Girandurukotte (CKDu endemic) and Kandy (CKDu non-endemic) to determine whether exposure to hantaviruses is a potential risk factor in patients with kidney disease. An indirect immunofluorescent antibody assay using two antigens, Thailand orthohantavirus-infected and recombinant N protein-expressing Vero E6 cells, were used for serodiagnosis. Participants' demographic and other socio-economic data were collected through a structured questionnaire. Fifty kidney disease patients and 270 controls from Kandy and 104 kidney disease patients and 242 controls from Girandurukotte were examined. Seropositivities were 50% and 17.4% in kidney patients and controls, respectively, in Girandurukotte, and they were 18% and 7% in Kandy. The odds of exposure to hantaviruses were higher for kidney disease patients than for controls in both Girandurukotte (OR:3.66, 95% CI:2.01 to 6.64) and Kandy (OR:2.64, 95% CI:1.07 to 6.54) in binary logistic regression models. According to statistical analysis, individuals exposed to hantaviruses had a higher risk of developing renal impairment. Therefore, hantavirus infection might be an important risk factor for development of kidney disease in Sri Lanka.


Assuntos
Infecções por Hantavirus/complicações , Infecções por Hantavirus/epidemiologia , Insuficiência Renal Crônica/virologia , Adulto , Proteínas do Capsídeo/imunologia , Estudos de Casos e Controles , Estudos Transversais , Doenças Endêmicas , Fazendeiros/estatística & dados numéricos , Feminino , Técnica Indireta de Fluorescência para Anticorpo , Geografia , Orthohantavírus , Infecções por Hantavirus/diagnóstico , Humanos , Masculino , Insuficiência Renal Crônica/epidemiologia , Fatores de Risco , Testes Sorológicos , Sri Lanka/epidemiologia , Proteínas do Core Viral/imunologia
14.
J Vet Med Sci ; 78(2): 221-30, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26412049

RESUMO

The applicability of the recombinant LipL32 for serodiagnosis of leptospiral infection in field rodents was assessed in this study. An immunodominant region of LipL32 was determined by monoclonal antibodies, and then, truncated LipL32 (tLipL32) was designed to contain the region (87-188th amino acid). The tLipL32 was compared between two recombinant expression hosts Escherichia coli and Pichia pastoris in ELISA. With field rat sera, tLipL32 expressed by P. pastoris (tLipL32p) had high antigenicity without background reactions, while tLipL32 expressed by E. coli (tLipL32e) showed high background reactions, which were reduced by pre-adsorption of sera with E. coli. To evaluate tLipL32-ELISA, field rat sera were tentatively divided into a Leptospira infection positive (12 sera) and a negative group (12 sera) based on the results from flaB gene PCR of kidney samples and WB with whole Leptospira cell. Consequently, the sensitivity of tLipL32p-ELISA for field rat sera was 83% . A similar result was obtained from tLipL32e-ELISA with adsorbed sera, (92%). However, sensitivity of tLipL32e-ELISA using sera without an adsorption treatment was 50%. Regardless of the expression host, tLipL32-ELISA had 100% specificity and sensitivity in experimentally infected laboratory rats. These results suggest that recombinant LipL32 expressed by P. pastoris is more applicable for serodiagnosis in field rats due to a lack of background reaction.


Assuntos
Proteínas da Membrana Bacteriana Externa , Leptospirose/diagnóstico , Lipoproteínas , Animais , Anticorpos Antibacterianos/imunologia , Anticorpos Monoclonais/imunologia , Proteínas da Membrana Bacteriana Externa/genética , Proteínas da Membrana Bacteriana Externa/imunologia , Ensaio de Imunoadsorção Enzimática , Mapeamento de Epitopos , Escherichia coli/genética , Feminino , Leptospira/imunologia , Lipoproteínas/genética , Lipoproteínas/imunologia , Fragmentos de Peptídeos/genética , Fragmentos de Peptídeos/imunologia , Pichia/genética , Ratos , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Testes Sorológicos
15.
Int Health ; 7(5): 348-53, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25549632

RESUMO

BACKGROUND: In Sri Lanka, one of the major challenges in rabies control is to manage the dog population and subsequently to protect people, especially young children, from dog bites. METHODS: In 2009, an educational-entertainment campaign called 'Rabies Edutainment 4 Kids' was introduced in the school curricula in rural Sri Lanka to improve practices on rabies prevention and pet care among school children, and to evaluate its effectiveness through pre- and post-tests. RESULTS: The level of rabies knowledge, attitude and practice among the pupils was dependent on their responses to a survey, and scores were significantly improved both among the study and control groups after the intervention. A lecture accompanied by a rabies awareness leaflet was much more effective in improving knowledge than the leaflet alone. The type of intervention and language used was significantly associated with the score increment (p<0.001). CONCLUSIONS: The threat of rabies to pupils in Sri Lanka would be reduced if they are given appropriate information on rabies prevention as a part of the school curricula. Close collaboration with local education offices is key to successful implementation of school-based rabies control programmes, which is, in turn, crucial to the eradication of rabies from Sri Lanka.


Assuntos
Educação em Saúde/organização & administração , Conhecimentos, Atitudes e Prática em Saúde , Raiva/prevenção & controle , População Rural , Serviços de Saúde Escolar/organização & administração , Animais , Criança , Cães , Feminino , Humanos , Masculino , Animais de Estimação , Vacina Antirrábica/administração & dosagem , Sri Lanka
16.
Genes Genet Syst ; 89(2): 71-80, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25224973

RESUMO

We examined genetic variation in black rats (the Rattus rattus complex) from Kandy District, Sri Lanka using mitochondrial cytochrome b (cytb, 1140 bp) and nuclear melanocortin 1 receptor (Mc1r, 954 bp) gene sequences together with database sequences. We confirmed the existence of two divergent mitochondrial lineages in Sri Lankan black rats, with genetic distance of 2.2% and estimated divergence time of 0.3 million years ago. Because one lineage is unique to the island and the other is closely related to R. rattus populations on the Indian subcontinent, two migration events of R. rattus from the subcontinent are inferred, one ancient and one recent. Mc1r analyses revealed 12 haplotypes among the Sri Lankan black rats. A median-joining network together with other available sequences separated the 12 haplotypes into two groups, one unique to the island and the other related to previously reported R. rattus sequences. Notably, most individuals possessed various combinations of both haplotype groups which had no association with the cytb clades. These results imply that old and new R. rattus lineages are now intermingled as a result of hybridization in Sri Lanka. Specimens of the lesser bandicoot rat (Bandicota bengalensis) collected from Sri Lanka (n = 24) were shown to have no genetic variability in the cytb sequence. Our results indicate that the two most abundant groups of commensal rats in Sri Lanka, black rats and lesser bandicoot rats, are the product of contrasting evolutionary histories on different timescales.


Assuntos
Núcleo Celular/genética , DNA Mitocondrial/genética , Variação Genética , Murinae/genética , Ratos/genética , Animais , Citocromos b/genética , Citocromos b/metabolismo , Evolução Molecular , Marcadores Genéticos , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Filogenia , Receptor Tipo 1 de Melanocortina/genética , Receptor Tipo 1 de Melanocortina/metabolismo , Análise de Sequência de DNA , Sri Lanka
17.
Glob Public Health ; 8(2): 159-73, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23205515

RESUMO

Religious leaders in Sri Lanka may have a high potential of contributing to HIV/AIDS prevention among the general public because of their social status. In order to assess their current HIV/AIDS-related knowledge and attitude and the possibility of becoming community advocates of HIV/AIDS prevention, we conducted a questionnaire survey among Buddhist, Hindu, Muslim and Christian leaders in Sri Lanka in 2009. There were limited correct responses about HIV/AIDS-related knowledge and attitudes toward people living with HIV/AIDS (PLHIV), and information regarding condoms, HIV testing and counselling were poorly understood. Although a condom was less acceptable as a part of HIV/AIDS prevention, they were willing to learn more about HIV/AIDS and expressed support for both PLHIV and HIV prevention activities. Their experiences, preparedness and willingness of HIV prevention activities were associated with age, knowledge and/or religious background. In conclusion, intensive and systematic learning opportunities should be provided to equip the religious leaders with overall HIV/AIDS knowledge to become key players for HIV/AIDS prevention in their communities.


Assuntos
Atitude Frente a Saúde/etnologia , Infecções por HIV/prevenção & controle , Conhecimentos, Atitudes e Prática em Saúde/etnologia , Religião e Medicina , Religião e Sexo , Adolescente , Adulto , Idoso , Budismo , Cristianismo , Comparação Transcultural , Feminino , Infecções por HIV/epidemiologia , Infecções por HIV/transmissão , Hinduísmo , Humanos , Islamismo , Liderança , Masculino , Pessoa de Meia-Idade , Prevalência , Sri Lanka/epidemiologia , Adulto Jovem
18.
J Virol Methods ; 193(1): 42-9, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23684845

RESUMO

Hantavirus is a causative agent of rodent-borne viral zoonoses, hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome. Seoul virus (SEOV) is a causative agent of urban and laboratory rat-associated HFRS worldwide. Surveillance of rodents has been done mainly by serological detection of hantavirus-specific antibodies by enzyme linked immunosorbent assay (ELISA) and immunofluorescent antibody assay (IFA). An immunochromatographic (ICG) test was developed with the N-terminal 103 amino acids of nucleocapsid protein of Hantaan virus expressed by Escherichia coli as an antigen to detect IgG antibody specific to hantavirus in sera from Rattus sp. animals. Antibody-detecting sensitivity of the ICG test was the same as that of ELISA and about 100-times higher than that of IFA. Overall sensitivities and specificities of the ICG test in comparison to ELISA and IFA for sera from 192 urban rats and 123 laboratory rats were 99.3% and 100%, respectively. Diluted whole blood samples without separation could be used for the ICG test. The ICG test enabled detection of antibodies to SEOV, Hantaan, Dobrava/Belgrade, and Thailand viruses, which are causative agents of HFRS throughout Eurasia. The ICG test is a rapid, simple and safe method for diagnosis of SEOV infection in rats.


Assuntos
Anticorpos Antivirais/sangue , Cromatografia de Afinidade/métodos , Testes Diagnósticos de Rotina/métodos , Infecções por Hantavirus/veterinária , Orthohantavírus/imunologia , Doenças dos Roedores/diagnóstico , Animais , Proteínas do Capsídeo/genética , Escherichia coli/genética , Feminino , Vírus Hantaan/genética , Vírus Hantaan/imunologia , Infecções por Hantavirus/imunologia , Imunoglobulina G/sangue , Ratos , Proteínas Recombinantes/genética , Doenças dos Roedores/imunologia , Sensibilidade e Especificidade , Tailândia , Proteínas do Core Viral/genética
20.
Jpn J Infect Dis ; 65(2): 157-61, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22446124

RESUMO

In Sri Lanka, leptospirosis is a notifiable disease. In addition to having a routine disease reporting system, Sri Lanka has implemented a hospital-based sentinel surveillance system since 2004. This report discusses the findings of a descriptive analysis of the sentinel surveillance data collected from 2005 to 2008. Of the 4,000 suspected leptospirosis cases, 46.9% and 26.8% were recorded from the Western and Sabaragamuwa provinces, respectively. Most of the individuals were male (83.5%), and approximately 45.6% were aged 30-49 years. Farmers accounted for 16.5%, and laborers for 16.1%; however, the occupation of nearly half (44.8%) of the study population was unknown. More than half (53.9%) of the individuals worked in paddy fields. Almost all had acute fever (98.8%), myalgia (92.9%), and headache (92.7%), but fewer had other related symptoms. Out of the 4,000 individuals, 2,496 (62.4%) underwent a laboratory test; however, the laboratory test results of only 1,445 (57.9%) and the microscopic agglutination results of 41 (2.8%) were available at the sentinel sites. Less than 2% of the reported individuals underwent prophylactic treatment. These findings will help enhance the ongoing efforts for controlling and preventing leptospirosis in Sri Lanka. Sentinel surveillance is a useful tool, but the data quality needs to be improved by supplementing the findings with adequate laboratory diagnosis data.


Assuntos
Leptospirose/epidemiologia , Vigilância de Evento Sentinela , Adolescente , Adulto , Distribuição por Idade , Idoso , Idoso de 80 Anos ou mais , Quimioprevenção/métodos , Criança , Pré-Escolar , Feminino , Hospitais , Humanos , Leptospirose/diagnóstico , Leptospirose/patologia , Masculino , Pessoa de Meia-Idade , Exposição Ocupacional , Distribuição por Sexo , Sri Lanka/epidemiologia , Adulto Jovem
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