RESUMO
The swamp type of the Asian water buffalo is assumed to have been domesticated by about 4000 years BP, following the introduction of rice cultivation. Previous localizations of the domestication site were based on mitochondrial DNA (mtDNA) variation within China, accounting only for the maternal lineage. We carried out a comprehensive sampling of China, Taiwan, Vietnam, Laos, Thailand, Nepal and Bangladesh and sequenced the mtDNA Cytochrome b gene and control region and the Y-chromosomal ZFY, SRY and DBY sequences. Swamp buffalo has a higher diversity of both maternal and paternal lineages than river buffalo, with also a remarkable contrast between a weak phylogeographic structure of river buffalo and a strong geographic differentiation of swamp buffalo. The highest diversity of the swamp buffalo maternal lineages was found in south China and north Indochina on both banks of the Mekong River, while the highest diversity in paternal lineages was in the China/Indochina border region. We propose that domestication in this region was later followed by introgressive capture of wild cows west of the Mekong. Migration to the north followed the Yangtze valley as well as a more eastern route, but also involved translocations of both cows and bulls over large distances with a minor influence of river buffaloes in recent decades. Bayesian analyses of various migration models also supported domestication in the China/Indochina border region. Coalescence analysis yielded consistent estimates for the expansion of the major swamp buffalo haplogroups with a credibility interval of 900 to 3900 years BP. The spatial differentiation of mtDNA and Y-chromosomal haplotype distributions indicates a lack of gene flow between established populations that is unprecedented in livestock.
Assuntos
Búfalos/genética , DNA Mitocondrial/genética , Genética Populacional , Cromossomo Y/genética , Animais , Animais Domésticos/genética , Ásia , Teorema de Bayes , Feminino , Fluxo Gênico , Haplótipos , Masculino , Modelos Genéticos , Filogeografia , Análise de Sequência de DNARESUMO
Due to human activity and a reduction in the size and quality of wetland habitats, populations of the Eastern sarus crane (Grus antigone sharpii) have declined dramatically across their range in Southeast Asia. Conservation efforts in Thailand have focused on reintroduction of the founders harboring the highest genetic diversity. One of the most important requirements to ensure the persistence of the reintroduced populations is a balanced sex ratio. In this study we tested three simple PCR-based methods which may be used for reliable sex identification in G. a. sharpii. The first method employs two combined primer sets based on a 0.6 kb EcoRI fragment (EE0.6). The second method is based on the intronic length polymorphism of the chromo-helicase DNA binding protein (CHD). The last technique relies on PCR-RFLP technique. The sex of six known and 24 unknown cranes were successfully identified by all three methods. These PCR-based sex identification methods are also useful for captive breeding management of G. a. sharpii.
Assuntos
Aves/genética , Aves/fisiologia , Reação em Cadeia da Polimerase/veterinária , Análise para Determinação do Sexo/veterinária , Animais , Sequência de Bases , Conservação dos Recursos Naturais , DNA/genética , Espécies em Perigo de Extinção , Análise para Determinação do Sexo/métodos , TailândiaRESUMO
The potential of macaque Macaca nemestrina leonina in Thailand to be infected by endemic arboviruses was assessed. The prevalence of antibodies of three arboviruses actively circulating in Thailand was determined by Plaque Reduction Neutralization assay procedures using samples from captive colonies in Northern Thailand. Out of 38 macaques, 9 (24%) presented reacting antibodies against dengue virus, 5 (13%) against Japanese encephalitis virus, and 4 (10%) against Chikungunya virus. Our results indicate that the northern pig-tailed macaque in Thailand can be infected by these arboviruses, inferring therefore that their virus specific vectors have bitten them. Given that, northern pig-tailed macaque represents an abundant population, living in close range to human or in peridomestic setting, they could play a role as potential reservoir host for arboviruses circulating in Thailand.
Assuntos
Infecções por Alphavirus/veterinária , Anticorpos Antivirais/sangue , Dengue/veterinária , Encefalite Japonesa/veterinária , Macaca nemestrina , Doenças dos Macacos/epidemiologia , Infecções por Alphavirus/epidemiologia , Infecções por Alphavirus/virologia , Animais , Vírus Chikungunya/isolamento & purificação , Dengue/epidemiologia , Dengue/virologia , Vírus da Dengue/isolamento & purificação , Vírus da Encefalite Japonesa (Espécie)/isolamento & purificação , Encefalite Japonesa/epidemiologia , Encefalite Japonesa/virologia , Doenças dos Macacos/virologia , Estudos Soroepidemiológicos , Tailândia/epidemiologia , Ensaio de Placa ViralRESUMO
The elephant endotheliotropic herpesvirus (EEHV) is now recognized as one of the main causes of death of young Asian elephants (Elephas maximus) in North American zoos. Its impact in wild and domestic elephant populations in Asia is not clearly understood. This article describes the first case of EEHV infection in Lao People's Democratic Republic of a 2.5-yr-old domestic male Asian elephant. Clinical signs and pathological findings reported here are consistent with previous infections in Asian elephant calves. Phylogenetic analyses showed 100% homology with other EEHV-1A strains identified in Asia, Europe, and North America. Contamination of the molecular assays was ruled out, because the DNA polymerase sequence identified in this study differed from the positive control by two base pairs.
Assuntos
Elefantes , Infecções por Herpesviridae/veterinária , Herpesviridae/classificação , Animais , Sequência de Bases , DNA Viral , Evolução Fatal , Herpesviridae/genética , Herpesviridae/isolamento & purificação , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/virologia , Laos/epidemiologia , Masculino , FilogeniaRESUMO
Background and Aim: Over the past two decades, the number of elephant camps in Thailand has increased considerably, and captive elephants have become more popular within the tourism industry. Tourist activities involving elephant exhibitions and trekking potentially affect animal health and welfare. This study aimed to investigate the relationships between a novel stress biomarker, fecal glucocorticoid metabolites (fGCM), and various factors (sex, age, weather season, tourist season, and elephant usage patterns), monitoring the fGCM concentration during and after trekking activities ceased. Materials and Methods: Fecal samples of 20 captive Asian elephants from two camps in Kanchanaburi Province were collected monthly for 1 year. The fGCM concentrations were measured using enzyme immunoassay and evaluated relative to individual demography, season, and tourist trekking activity. The mean differences of fGCMs concentrations were compared by analysis of variance and t-test statistics according to data types with p<0.5. Results: Significant differences in mean fGCM concentrations were found between age categories (p=0.001), trekking and non-trekking animals (p=0.039), and during and after trekking (p=0.023). The mean fGCM concentration of elephants aged during 0-44 years (136.7 ng/g) was significantly higher than for animals over 44 years old (107.7 ng/g), and the elephant trekking group (144.9 ng/g) was significantly higher than the other group (124.7 ng/g). Within the trekking group, the mean fGCM concentrations gradually declined to 129.13 ng/g within 8 months of trekking cessation. Conclusion: Elephant's ages and activities co-influenced the variance of fGCM concentrations. In addition, permanent tourist activity, especially trekking, can increase elephant stress. This study's findings can be applied to the health status monitoring of captive elephants and result in improved animal welfare.
RESUMO
The recombinant, modified leucine-rich repeat protein rhKU_Sej_LRR_2271 has been suggested as a candidate for leptospiral vaccine development since it was predicted to be a transmembrane protein containing leucine-rich repeat motifs and immunogenic epitopes. The immunogenic epitopes showed binding affinities with lower IC50 values than peptides of known antigenic proteins, e.g., LipL32. Moreover, this protein was immunoreactive with hyperimmune sera against several serovars. In this study, we aimed to develop a lateral flow strip test using the rhKU_Sej_LRR_2271 protein for the detection of anti-leptospiral IgG in dogs. The lateral flow assay was performed with 184 dog plasma samples and evaluated with a culture method, 16S ribosomal RNA gene (rss) analysis real-time PCR, and LipL32 ELISA. The culture method failed to detect leptospires in the dog blood samples. Six of nine symptomatic dogs gave positive results with the real-time PCR assay. The lateral flow assay and LipL32 ELISA gave positive results with 59 and 50 dogs, respectively. The sensitivity, specificity, and accuracy of the rhKU_Sej_LRR_2271 lateral flow strip test were 70.00, 82.09, and 78.80%, respectively, when compared with LipL32 ELISA. There was a significant association between the LipL32 ELISA and the rhKU_Sej_LRR_2271 lateral flow assay. The rhKU_Sej_LRR_2271 lateral flow strip test has therefore demonstrated a good potential to detect anti-leptospiral IgG in dogs.
RESUMO
Nonstructural 3ABC protein of foot and mouth disease virus (FMDV) was widely used to differentiate vaccinated from natural FMDV-infected animals. 3ABC is a polyprotein which is auto-processed to 3A, three copies of 3B and 3C(pro) by 3C(pro) protease. The 3ABC gene was cloned and expressed in Escherichia coli as native or mutated 3ABC (mu3ABC) forms. Cysteine residues 142 and 163 of the catalytic triad within the 3C(pro) of mu3ABC were changed to serine and glycine, respectively, to inhibit its protease activity. Both native and mutated 3ABC ORFs were cloned into BamHI and HindIII restriction sites of an expression vector, pQE80L. The expression of the recombinant native 3ABC and mu3ABC genes in E. coli BL21 was induced with 0.2mM isopropyl-beta-d-thiogalactopyranoside at 37 °C for 5h. SDS-PAGE and Western blot analysis revealed that the full length 3ABC was present in the lysate from mu3ABC but not native 3ABC transformed cells. The recombinant mu3ABC was expressed mainly in the inclusion body and presented as monomer and dimer. In addition, the mu3ABC reacted strongly with a convalescent serum from a natural FMDV-infected cattle but very weakly with a serum from vaccinated cattle. This study clearly demonstrates that successful expression of the full length 3ABC occurs only when the protease active sites within the 3C(pro) were completely abolished. This information would accelerate in house development of the 3ABC-based diagnostic test that can distinguish between vaccinated and FMDV-infected animals.
Assuntos
Clonagem Molecular , Escherichia coli/genética , Vírus da Febre Aftosa/genética , Proteínas não Estruturais Virais/genética , Sequência de Aminoácidos , Animais , Bovinos , Febre Aftosa/genética , Expressão Gênica , Dados de Sequência Molecular , Mutação , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificação , Proteínas não Estruturais Virais/isolamento & purificaçãoRESUMO
Abstract. The objectives of this study were to clone the conserved region of leptospiral immunoglobulin-like protein (lig) gene and evaluate the utility of the recombinant Lig as an ELISA antigen for detection of leptospiral antibodies in canine sera. Leptospira kirschneri serovar Grippotyposa strain Moskva V was chosen to be a target for cloning the conserved region of Lig gene. This assay was evaluated with canine sera (n = 91) that were MAT-negative (< 1:100 dilution) and sera (n = 103) that were MAT-positive (> or = 1:100 dilution) using 24 serovars. The ELISA showed a relative sensitivity as compared to MAT of 84.5% whereas the specificity was 76.9%. This assay is simple and can be routinely prepared in large amounts. It was concluded that the GST.Lig recombinant protein-based ELISA could be used as a screening test for serodiagnosis of canine leptospirosis with also for confirmation of MAT-positive test results.
Assuntos
Doenças do Cão/microbiologia , Leptospira/imunologia , Leptospira/isolamento & purificação , Leptospirose/veterinária , Testes de Aglutinação/métodos , Animais , Anticorpos Antibacterianos/análise , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/isolamento & purificação , Clonagem Molecular , Doenças do Cão/sangue , Doenças do Cão/diagnóstico , Cães , Ensaio de Imunoadsorção Enzimática/métodos , Ensaio de Imunoadsorção Enzimática/veterinária , Imunoglobulinas/análise , Imunoglobulinas/sangue , Imunoglobulinas/imunologia , Leptospirose/sangue , Leptospirose/diagnóstico , Leptospirose/imunologia , Sensibilidade e Especificidade , TailândiaRESUMO
Four Asian elephants were confirmed to be infected with Mycobacterium tuberculosis by bacterial culture, other diagnostic procedures, and sequencing of 16S-23S rDNA internal transcribed spacer region, 16S rRNA, and gyrase B gene sequences. Genotyping showed that the infectious agents originated from 4 sources in Thailand. To identify infections, a combination of diagnostic assays is essential.
Assuntos
Animais Domésticos/microbiologia , Elefantes/microbiologia , Mycobacterium tuberculosis/isolamento & purificação , RNA Ribossômico 23S/análise , Tuberculose/veterinária , Animais , Proteínas de Bactérias/genética , DNA Girase/genética , DNA Espaçador Ribossômico , Mycobacterium tuberculosis/genética , Reação em Cadeia da Polimerase , RNA Bacteriano/análise , RNA Ribossômico 16S/análise , Alinhamento de Sequência , Tailândia , Tuberculose/diagnóstico , Tuberculose/microbiologiaRESUMO
A CD30 ligand (CD30L, CD153) is a type II membrane-associated glycoprotein belonging to the TNF family. To illustrate the potential role of CD30L in CD4(+) Th1 cell responses, we investigated the fate of Ag-specific CD4(+) T cells in CD30L-deficient (CD30L(-/-)) mice after Mycobacterium bovis bacillus Calmette-Guérin (BCG) infection. The number of bacteria was significantly higher in organs of CD30L(-/-) mice than in wild-type (WT) mice 4 wk postinfection. The numbers of purified protein derivative- or Ag85B-specific-IFN-gamma-producing-CD4(+) T cells in spleen, lung, or peritoneal exudate cells were significantly fewer in CD30L(-/-) mice than in WT mice. During the infection, CD30L was expressed mainly by CD44(+)CD3(+)CD4(+) T cells but not by CD3(+)CD8(+) T cells, B cells, dendritic cells, or macrophages. Costimulation with agonistic anti-CD30 mAb or coculturing with CD30L-transfected P815 cells restored IFN-gamma production by CD4(+) T cells from BCG-infected CD30L(-/-) mice. Coculturing with CD30L(+/+)CD4(+) T cells from BCG-infected WT mice also restored the number of IFN-gamma(+)CD30L(-/-)CD4(+) T cells. When transferred into the CD30L(+/+) mice, Ag-specific donor CD30L(-/-) CD4(+) T cells capable of producing IFN-gamma were restored to the compared level seen in CD30L(+/+) CD4(+) T cells on day 10 after BCG infection. When naive CD30L(+/+) T cells were transferred into CD30L(-/-) mice, IFN-gamma-producing-CD4(+) Th1 cells of donor origin were normally generated following BCG infection, and IFN-gamma-producing-CD30L(-/-)CD4(+) Th1 cells of host origin were partly restored. These results suggest that CD30L/CD30 signaling executed by CD30(+) T-CD30L(+) T cell interaction partly play a critical role in augmentation of Th1 response capable of producing IFN-gamma against BCG infection.
Assuntos
Ligante CD30/fisiologia , Comunicação Celular/imunologia , Antígeno Ki-1/fisiologia , Infecções por Mycobacterium/imunologia , Transdução de Sinais/imunologia , Subpopulações de Linfócitos T/imunologia , Células Th1/imunologia , Células Th1/microbiologia , Animais , Ligante CD30/biossíntese , Ligante CD30/deficiência , Ligante CD30/genética , Linhagem Celular Tumoral , Células Cultivadas , Técnicas de Cocultura , Predisposição Genética para Doença , Interferon gama/antagonistas & inibidores , Interferon gama/biossíntese , Antígeno Ki-1/biossíntese , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Infecções por Mycobacterium/patologia , Infecções por Mycobacterium/prevenção & controle , Mycobacterium bovis/crescimento & desenvolvimento , Mycobacterium bovis/imunologia , Subpopulações de Linfócitos T/metabolismo , Subpopulações de Linfócitos T/patologia , Células Th1/metabolismo , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/patologia , Tuberculose Pulmonar/prevenção & controleRESUMO
Zoonotic pathogens such as arboviruses have comprised a significant proportion of emerging infectious diseases in humans. The role of wildlife species as reservoirs for arboviruses is poorly understood, especially in endemic areas such as Southeast Asia. This study aims to determine the exposure history of different macaque species from national parks in Thailand to mosquito-borne flaviviruses and alphavirus by testing the serum samples collected from 25 northern pigtailed macaques, 33 stump-tailed macaques, and 4 long-tailed macaques for the presence of antibodies against dengue, Zika, and chikungunya viruses by plaque reduction neutralization assay. Specific neutralizing antibodies against Dengue virus (DENV1-4) and Zika virus (ZIKV) were mainly found in stump-tailed macaques, whereas neutralizing antibody titers were not detected in long-tailed macaques and pigtailed macaques as determined by 90% plaque reduction neutralization assay (PRNT90). One long-tailed macaque captured from the south of Thailand exhibited antibody titers against chikungunya virus (CHIKV), suggesting enzootic of this virus to nonhuman primates (NHPs) in Thailand. Encroachment of human settlements into the forest has increased the interface that exposes humans to zoonotic pathogens such as arboviruses found in monkeys. Nonhuman primates living in different regions of Thailand showed different patterns of arboviral infections. The presence of neutralizing antibodies among wild monkeys in Thailand strongly suggests the existence of sylvatic cycles for DENV, ZIKV, and CHIKV in Thailand. The transmission of dengue, Zika, and chikungunya viruses among wild macaques may have important public health implications.
Assuntos
Febre de Chikungunya/epidemiologia , Vírus Chikungunya/imunologia , Culicidae/virologia , Vírus da Dengue/imunologia , Dengue/epidemiologia , Infecção por Zika virus/epidemiologia , Zika virus/imunologia , Animais , Anticorpos Neutralizantes , Anticorpos Antivirais/sangue , Febre de Chikungunya/virologia , Dengue/virologia , Feminino , Geografia , Humanos , Macaca , Masculino , Mosquitos Vetores/virologia , Estudos Soroepidemiológicos , Tailândia/epidemiologia , Infecção por Zika virus/virologiaRESUMO
In this study, a first food-grade mucosal vaccine against leptospirosis was developed without the use of antibiotic resistance gene. This expression system is based on a food-grade host/vector system of Lactobacillus plantarum and a new vaccine candidate antigen, a leucine-rich repeat (LRR) protein of Leptospira borgpetersenii. The LRR of interest from serovar Sejroe is encoded by two overlapping genes and these genes were fused together by site-directed mutagenesis. The mutant gene thus obtained could be successfully expressed in this system as was shown by western blot analysis and liquid chromatography-mass spectrometry (LC-MS/MS) analysis. In addition, this analysis showed that the mutant LRR protein fused to a homologous signal peptide of L. plantarum could be exported to the cell surface as a result of the native LPXAG motif of the heterologous LRR protein, which presumably is responsible for anchoring the protein to the cell wall of L. plantarum. This new strategy could be an essential tool for further studies of leptospirosis mucosal vaccine delivery.
RESUMO
Porcine circovirus type 3 (PCV3) has recently been detected in pigs worldwide, with similar clinical manifestations to porcine circovirus-associated disease (PCVAD) from porcine circovirus type 2 (PCV2) infection. Here, we report the identification and molecular epidemiology of PCV3 in swine in Thailand from clinical samples retrieved from 2006 to 2017. The epidemiological data revealed co-infection with PCV2, PRRSV, and PCV2/PRRSV was common in our samples. Circulating PCV3 from this study shared a high similarity of nucleotide and deduced amino acid sequences of the partial capsid gene (96.7%-100% and 96.7%-100% respectively), indicated the genetic stability of PCV3 in Thailand. Phylogenetic analysis based on the capsid gene revealed scatter clustering with current PCV3 having no relation to the geographical origin of the virus strains. In this retrospective study, results have demonstrated that PCV3 has spread extensively within Thai swine from as early as 2006 and may also be involved in PRDC and PCVAD.
Assuntos
Infecções por Circoviridae/virologia , Circovirus/genética , Doenças dos Suínos/virologia , Sequência de Aminoácidos , Animais , Proteínas do Capsídeo/genética , Infecções por Circoviridae/epidemiologia , Circovirus/isolamento & purificação , Coinfecção , Geografia , Epidemiologia Molecular , Filogenia , Estudos Retrospectivos , Suínos , Doenças dos Suínos/epidemiologia , Tailândia/epidemiologiaRESUMO
We studied the occurrence of swine influenza virus (SIV) infection in piglets with respiratory symptoms resembling porcine respiratory disease complex (PRDC). A total of 106 samples including nasal swab and lung suspension from sick piglets were collected from 30 farms of medium size in the central and eastern parts of Thailand from August 2006 to February 2007. Samples were inoculated onto Mardin-Darby Canine Kidney (MDCK) cells and SIV infection was confirmed by immunofluorescent assay (IFA) and reverse transcriptase polymerase chain reaction (RT-PCR) specific for M gene. Of 106 samples, 3 pigs from 3 different farms were found to be SIV positive on all assays. The positive samples were further identified by RT-PCR as H3N2 subtype using specific primers for hemagglutinin (HA) and neuraminidase (NA) genes. SIV infection was found in 2.8% of swine suffering from respiratory distress suggesting SIV may not be the major pathogen for PRDC in the central and eastern Thailand. SIV was present in 3 of 30 farms (10%) indicating the prevalence of SIV in these regions is considerable. Since pigs are vulnerable to infection from both human and avian influenza viruses and interspecies transmission between humans and swine occurs sporadically, it is essential to continue surveillance and monitoring of SIV infection in the swine population.
Assuntos
Vírus da Influenza A/isolamento & purificação , Infecções por Orthomyxoviridae/veterinária , Infecções Respiratórias/veterinária , Doenças dos Suínos/epidemiologia , Animais , Linhagem Celular , Primers do DNA , Cães , Imunofluorescência , Vírus da Influenza A Subtipo H3N2/genética , Vírus da Influenza A Subtipo H3N2/isolamento & purificação , Vírus da Influenza A/genética , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/virologia , Prevalência , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Suínos , Doenças dos Suínos/virologia , Tailândia/epidemiologiaRESUMO
We presented the powerful techniques for species identification using the short amplicon of mitochondrial cytochrome b gene sequence. Two faecal samples and one single hair sample of the Asian tapir were tested using the new cytochrome b primers. The results showed a high sequence similarity with the mainland Asian tapir group. The comparative sequence analysis of the reserved wild mammals in Thailand and the other endangered mammal species from Southeast Asia comprehensibly verified the potential of our novel primers. The forward and reverse primers were 94.2 and 93.2%, respectively, by the average value of the sequence identity among 77 species sequences, and the overall mean distance was 35.9%. This development technique could provide rapid, simple, and reliable tools for species confirmation. Especially, it could recognize the problematic biological specimens contained less DNA material from illegal products and assist with wildlife crime investigation of threatened species and related forensic casework.
Assuntos
Código de Barras de DNA Taxonômico/métodos , Espécies em Perigo de Extinção , Genes Mitocondriais , Perissodáctilos/genética , Animais , Sequência de Bases , Citocromos b/genética , Primers do DNA , Mamíferos/classificação , Mamíferos/genética , Perissodáctilos/classificação , Filogenia , Alinhamento de Sequência , TailândiaRESUMO
Immunolocalization of inhibin-α and inhibin/activin ßA and ßB subunits in the testes of Asian elephant was determined. Testicular sections were immunostained with polyclonal antisera against inhibin subunit-α and inhibin/activin ßA and ßB using the avidin-biotin-peroxidase complex method. Positive immunostaining against inhibin-α subunit was strongly present in Sertoli cells, and positive immunostaining for the inhibin/activin ßA and ßB subunits was observed in both Sertoli and Leydig cells. These results indicated that while Sertoli cells are the predominant source of inhibin and activin secretions in the testes of adult male Asian elephant, Leydig cells are a source of activin but not inhibin.
Assuntos
Ativinas/análise , Elefantes , Subunidades beta de Inibinas/análise , Animais , Células Intersticiais do Testículo/metabolismo , Masculino , Células de Sertoli/metabolismoRESUMO
Several simian adenoviruses (AdVs) have been detected and isolated in various species of non-human primates with the goals of monitoring the health of wildlife and investigating their potential for zoonotic disease transmission. Here, we provide evidence of AdV infection in wild Assamese macaques (Macaca assamensis assamensis) at Phu Khieo Wildlife Sanctuary, Thailand, based on polymerase chain reaction of non-invasively collected fecal samples. Eight out of 110 fecal samples (7.3%), or five out of 87 monkeys (5.7%), showed evidence of AdV infection. All infected individuals were infants or juveniles. Phylogenetic analysis based on the sequence of hexon and polymerase genes revealed two different AdV genotypes. One genotype clustered in the human AdV-G group, while another showed 100% identity with previously reported AdVs of captive Chinese rhesus macaques (Macaca mulatta), which may be tentatively classified as a new species of AdV in non-human primates while awaiting further supporting evidence.
Assuntos
Infecções por Adenoviridae/veterinária , Adenoviridae/genética , Macaca , Doenças dos Macacos/virologia , Adenoviridae/classificação , Adenoviridae/isolamento & purificação , Infecções por Adenoviridae/epidemiologia , Infecções por Adenoviridae/virologia , Animais , Proteínas do Capsídeo/genética , Feminino , Masculino , Doenças dos Macacos/epidemiologia , Filogenia , Reação em Cadeia da Polimerase/veterinária , Prevalência , Tailândia/epidemiologiaRESUMO
To clarify the reproductive cycle of female Rusa deer (Rusa timorensis), the fecal concentrations of progesterone and 17ß-estradiol metabolites were measured. Fecal samples were collected on a weekly basis for one year (between October, 2012 and September, 2013) from five healthy adult hinds in Thailand. At the beginning of the study, three hinds were pregnant. Two hinds delivered one healthy offspring, and one hind delivered a stillborn calf. The mating period of Rusa hinds in Thailand is from November to April. In pregnant hinds, fecal progesterone metabolite concentration was high in late pregnancy and abruptly declined to the baseline around parturition, suggesting that the placenta secretes a large amount of progesterone. Fecal 17ß-estradiol metabolite concentration remained elevated around the day of parturition. Both concentrations of fecal progesterone and 17ß-estradiol metabolites in non-lactating hinds were significantly higher than those in lactating hinds, indicating that ovarian activity of lactating hinds is suppressed by the suckling stimulus of fawn during lactation. The present study demonstrated that monitoring of fecal steroid hormones is useful method for assessing ovarian function in this species.
Assuntos
Cervos/fisiologia , Estradiol/metabolismo , Ciclo Estral/metabolismo , Fezes/química , Progesterona/metabolismo , Animais , Feminino , Lactação/fisiologia , Período Pós-Parto/fisiologia , Gravidez , TailândiaRESUMO
Sarcocystis species are heteroxenous cyst-forming coccidian protozoan parasites with a wide host range, including rodents. In this study, Sarcocystis spp. samples were isolated from Bandicota indica, Rattus argentiventer, R. tiomanicus and R. norvegicus across five provinces of Thailand. Two major groups of Sarcocystis cysts were determined in this study: large and small cysts. By sequence comparisons and phylogenetic analyses based on the partial sequences of 28S ribosomal DNA, the large cysts showed the highest identity value (99%) with the S. zamani in GenBank database. While the small cysts could be divided into 2 groups of Sarcocystis: S. singaporensis and presupposed S. zuoi. The further analysis on 18S rDNA supported that the 2 isolates (S2 and B6 no.2) were as identified as S. singaporensis shared a high sequence identity with the S. singaporensis in GenBank database and the unidentified Sarcocystis (4 isolates, i.e., B6 no.10, B6 no.12, B10 no.4 and B10 no.7) showed 96.3-99.5% identity to S. zuoi as well as high distinct identity from others Sarcocystis spp. (≤93%). The result indicated that these four samples should be S. zuoi. In this study, we provided complete sequence of internal transcribed spacer 1 (ITS1), 5.8S rDNA and internal transcribed spacer 2 (ITS2) of these three Sarcocystis species and our new primer set could be useful to study the evolution of Sarcocystis.
Assuntos
Doenças dos Roedores/parasitologia , Sarcocystis/genética , Sarcocistose/veterinária , Animais , DNA de Protozoário/genética , DNA de Protozoário/isolamento & purificação , Filogenia , RNA de Protozoário/genética , RNA Ribossômico 28S/genética , Doenças dos Roedores/epidemiologia , Roedores , Sarcocystis/classificação , Sarcocystis/isolamento & purificação , Sarcocistose/epidemiologia , Sarcocistose/parasitologia , Tailândia/epidemiologiaRESUMO
Leptospirosis is an important zoonotic disease, and the major outbreak of this disease in Thailand in 1999 was due largely to the Leptospira borgpetersenii serovar Sejroe. Identification of the leucine-rich repeat (LRR) LBJ_2271 protein containing immunogenic epitopes and the discovery of the LBJ_2271 ortholog in Leptospira serovar Sejroe, KU_Sej_R21_2271, led to further studies of the antigenic immune properties of KU_Sej_LRR_2271. The recombinant hybrid (rh) protein was created and expressed from a hybrid PCR fragment of KU_Sej_R21_2271 fused with DNA encoding the LBJ_2271 signal sequence for targeting protein as a membrane-anchoring protein. The fusion DNA was cloned into pET160/GW/D-TOPO® to form the pET160_hKU_R21_2271 plasmid. The plasmid was used to express the rhKU_Sej_LRR_2271 protein in Escherichia coli BL21 Star™ (DE3). The expressed protein was immunologically detected by Western blotting and immunoreactivity detection with hyperimmune sera, T cell epitope prediction by HLA allele and epitope peptide binding affinity, and potential T cell reactivity analysis. The immunogenic epitopes of the protein were evaluated and verified by HLA allele and epitope peptide complex structure molecular docking. Among fourteen best allele epitopes of this protein, binding affinity values of 12 allele epitopes remained unchanged compared to LBJ_2271. Two epitopes for alleles HLA-A0202 and -A0301 had higher IC50 values, while T cell reactivity values of these peptides were better than values from LBJ_2271 epitopes. Eight of twelve epitope peptides had positive T-cell reactivity scores. Although the molecular docking of two epitopes, 3FPLLKEFLV11/47FPLLKEFLV55 and 50KLSTVPEGV58, into an HLA-A0202 model revealed a good fit in the docked structures, 50KLSTVPEGV58 and 94KLSTVPEEV102 are still considered as the proteins' best epitopes for allele HLA-A0202. The results of this study showed that rhKU_Sej_LRR_2271 protein contained natural immunological properties that should be further examined with respect to antigenic immune stimulation for vaccine development to prevent prevalent leptospiral serovar infection in Thailand.