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1.
Genomics ; 114(6): 110501, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-36270383

RESUMO

BACKGROUND: The iconic giant panda (Ailuropoda melanoleuca), as both a flagship and umbrella species endemic to China, is a world famous symbol for wildlife conservation. The giant panda has several specific biological traits and holds a relatively small place in evolution. A high-quality genome of the giant panda is key to understanding the biology of this vulnerable species. FINDINGS: We generated a 2.48-Gb chromosome-level genome (GPv1) of the giant panda named "Jing Jing" with a contig N50 of 28.56 Mb and scaffold N50 of 134.17 Mb, respectively. The total length of chromosomes (n = 21) was 2.39-Gb, accounting for 96.4% of the whole genome. Compared with the previously published four genomes of the giant panda, our genome is characterized by the highest completeness and the correct sequence orientation. A gap-free and 850 kb length of immunoglobulin heavy-chain gene cluster was manually annotated in close proximity to the telomere of chromosome 14. Additionally, we developed an algorithm to predict the centromere position of each chromosome. We also constructed a complete chromatin structure for "Jing Jing", which includes inter-chromosome interaction pattern, A/B compartment, topologically associated domain (TAD), TAD-clique and promoter-enhancer interaction (PEI). CONCLUSIONS: We presented an improved chromosome-level genome and complete chromatin structure for the giant panda. This is a valuable resource for the future genetic and genomic studies on giant panda.


Assuntos
Ursidae , Animais , Ursidae/genética , Genômica , China , Cromossomos/genética , Cromatina
2.
Cryobiology ; 94: 95-99, 2020 06.
Artigo em Inglês | MEDLINE | ID: mdl-32304703

RESUMO

Sperm cryopreservation is an essential approach for assisted reproduction and genetic resources conservation in captive giant pandas. Cryopreservation, however, leads to a significant decrease in sperm quality and, consequently, a low fertilization rate. Therefore, it is mandatory to disclose more suitable and efficient freezing strategies for sperm cryopreservation. In the present study, we compared for the first time the performance of two commercial freeze extender (INRA96 versus TEST) freezing methods on post-thawed semen quality. Semen cryopreserved with the INRA96 showed better total motility (73.00 ± 4.84% vs 57.56 ± 3.60%, P < 0.001), membrane integrity (60.92 ± 2.27% vs 40.53 ± 2.97%, P < 0.001) and acrosome integrity (90.39 ± 2.74% vs 84.26 ± 4.27%, P < 0.05) than stored with TEST. There was no significant difference in DNA integrity after thawing between the two extenders (95.69 ± 3.60% vs 94.26 ± 4.84%). In conclusion, the INRA96 method showed to be better for giant panda sperm cryopreservation and should therefore be recommended for use in order to increase success of artificial insemination.


Assuntos
Criopreservação/métodos , Crioprotetores/farmacologia , Preservação do Sêmen/métodos , Sêmen , Ursidae , Animais , Masculino , Análise do Sêmen , Espermatozoides/efeitos dos fármacos
3.
Korean J Parasitol ; 52(1): 21-6, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-24623877

RESUMO

Dirofilaria immitis (heartworm) infections affect domestic dogs, cats, and various wild mammals with increasing incidence in temperate and tropical areas. More sensitive antibody detection methodologies are required to diagnose asymptomatic dirofilariasis with low worm burdens. Applying current transcriptomic technologies would be useful to discover potential diagnostic markers for D. immitis infection. A filarial homologue of the mammalian translationally controlled tumor protein (TCTP) was initially identified by screening the assembled transcriptome of D. immitis (DiTCTP). A BLAST analysis suggested that the DiTCTP gene shared the highest similarity with TCTP from Loa loa at protein level (97%). A histidine-tagged recombinant DiTCTP protein (rDiTCTP) of 40 kDa expressed in Escherichia coli BL21 (DE3) showed immunoreactivity with serum from a dog experimentally infected with heartworms. Localization studies illustrated the ubiquitous presence of rDiTCTP protein in the lateral hypodermal chords, dorsal hypodermal chord, muscle, intestine, and uterus in female adult worms. Further studies on D. immitis-derived TCTP are warranted to assess whether this filarial protein could be used for a diagnostic purpose.


Assuntos
Antígenos de Helmintos/genética , Antígenos de Helmintos/isolamento & purificação , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/isolamento & purificação , Dirofilaria immitis/genética , Estruturas Animais/química , Animais , Anticorpos Anti-Helmínticos/sangue , Antígenos de Helmintos/química , Antígenos de Helmintos/imunologia , Biomarcadores Tumorais/química , Biomarcadores Tumorais/imunologia , Clonagem Molecular , Dirofilaria immitis/química , Dirofilaria immitis/imunologia , Modelos Animais de Doenças , Cães , Escherichia coli/genética , Expressão Gênica , Dados de Sequência Molecular , Peso Molecular , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/imunologia , Proteínas Recombinantes de Fusão/isolamento & purificação , Análise de Sequência de DNA , Proteína Tumoral 1 Controlada por Tradução
4.
Vet Res ; 44: 3, 2013 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-23331513

RESUMO

The heartworm Dirofilaria immitis is the causative agent of cardiopulmonary dirofilariosis in dogs and cats, which also infects a wide range of wild mammals and humans. The complex life cycle of D. immitis with several developmental stages in its invertebrate mosquito vectors and its vertebrate hosts indicates the importance of miRNA in growth and development, and their ability to regulate infection of mammalian hosts. This study identified the miRNA profiles of D. immitis of zoonotic significance by deep sequencing. A total of 1063 conserved miRNA candidates, including 68 anti-sense miRNA (miRNA*) sequences, were predicted by computational methods and could be grouped into 808 miRNA families. A significant bias towards family members, family abundance and sequence nucleotides was observed. Thirteen novel miRNA candidates were predicted by alignment with the Brugia malayi genome. Eleven out of 13 predicted miRNA candidates were verified by using a PCR-based method. Target genes of the novel miRNA candidates were predicted by using the heartworm transcriptome dataset. To our knowledge, this is the first report of miRNA profiles in D. immitis, which will contribute to a better understanding of the complex biology of this zoonotic filarial nematode and the molecular regulation roles of miRNA involved. Our findings may also become a useful resource for small RNA studies in other filarial parasitic nematodes.


Assuntos
Dirofilaria immitis/genética , MicroRNAs/genética , Animais , Dirofilariose/parasitologia , Doenças do Cão/parasitologia , Cães , Feminino , Sequenciamento de Nucleotídeos em Larga Escala/veterinária , Masculino , MicroRNAs/metabolismo , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/veterinária , Análise de Sequência de RNA/veterinária
5.
Microorganisms ; 11(7)2023 Jul 20.
Artigo em Inglês | MEDLINE | ID: mdl-37513017

RESUMO

Feline panleukopenia (FPL) is a highly contagious acute infectious disease caused by feline parvovirus (FPV). FPV has also been found in giant pandas with clinical signs of vomiting and mild diarrhea, posing a threat to this vulnerable species. Cleaning and disinfection may be one of the most efficacious ways to prevent FPV spread in the habitat of giant pandas. This study evaluated the inactivation effect of peracetic acid (PAA), povidone-iodine (PVP-I), glutaral and deciquam solution (JM) and Virkon S. The tissue culture infective dose (TCID50) assay indicated that the virus may be totally inactivated by JM, PAA and Virkon S. Meanwhile, the hemagglutination (HA) assay showed a high inactivation efficiency of PAA and Virkon S. The analysis of Western blot revealed that PAA, Virkon S and JM can inhibit the structural protein synthesis. Taken together, our findings demonstrated that PAA could rapidly and efficiently inactivate FPV, representing an efficacious disinfectant for FPV control.

6.
Front Immunol ; 14: 1237630, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37662912

RESUMO

Represented by feline panleukopenia virus (FPV) and canine parvovirus (CPV), the species carnivore protoparvovirus 1 has a worldwide distribution through continuous ci13rculation in companion animals such as cats and dogs. Subsequently, both FPV and CPV had engaged in host-to-host transfer to other wild animal hosts of the order Carnivora. In the present study, we emphasized the significance of cross-species transmission of parvoviruses with the isolation and characterization of an FPV from giant panda displaying severe and fatal symptoms. The isolated virus, designated pFPV-sc, displayed similar morphology as FPV, while phylogenetic analysis indicated that the nucleotide sequence of pFPV-sc clades with Chinese FPV isolates. Despite pFPV-sc is seemingly an outcome of a spillover infection event from domestic cats to giant pandas, our study also provided serological evidence that FPV or other parvoviruses closely related to FPV could be already prevalent in giant pandas in 2011. Initiation of host transfer of pFPV-sc is likely with association to giant panda transferrin receptor (TfR), as TfR of giant panda shares high homology with feline TfR. Strikingly, our data also indicate that pFPV-sc can infect cell lines of other mammal species, including humans. To sum up, observations from this study shall promote future research of cross-host transmission and antiviral intervention of Carnivore protoparvovirus 1, and necessitate surveillance studies in thus far unacknowledged potential reservoirs.


Assuntos
Vírus da Panleucopenia Felina , Ursidae , Humanos , Gatos , Animais , Cães , Vírus da Panleucopenia Felina/genética , Filogenia , Animais Selvagens , Tropismo
7.
Front Microbiol ; 14: 1117135, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36922967

RESUMO

Canine parvovirus (CPV) and Canine distemper virus (CDV) can cause fatal diseases in giant panda (Ailuropoda melanoleuca). The main capsid protein of CPV VP2 can be self-assembled to form virus-like particles (VLPs) in vitro, which is of great significance for potential vaccine development. In the present study, we remodeled the VP2 protein of a giant panda-derived CPV, where the major CDV F and N epitopes were incorporated in the N-terminal and loop2 region in two combinations to form chimeric VLPs. The reactivity ability and morphology of the recombinant proteins were confirmed by Western blot, hemagglutination (HA) test and electron microscopy. Subsequently, the immunogenicity of the VLPs was examined in vivo. Antigen-specific antibodies and neutralizing activity were measured by ELISA, hemagglutination inhibition (HI) test and serum neutralization test (SNT), respectively. In addition, antigen specific T cell activation were determined in splenic lymphocytes. The results indicated that the VLPs displayed good reaction with CDV/CPV antibodies, and the heterologous epitopes do not hamper solubility or activity. The VLPs showed decent HA activity, and resembled round-shaped particles with a diameter of 22-26 nm, which is identical to natural virions. VLPs could induce high levels of specific antibodies to CPV and CDV, shown by the indication of neutralizing antibodies in both VP2N and VP2L VLPs group. In addition, splenic lymphocytes of mice immunized with VLPs could proliferate rapidly after stimulation by specific antigen. Taken together, the CPV VP2 VLPs or chimeric VLPs are highly immunogenic, and henceforth could function as CPV/CDV vaccine candidates for giant pandas.

8.
In Vitro Cell Dev Biol Anim ; 59(7): 550-563, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37639049

RESUMO

Conservation of genetic resources is an important way to protect endangered species. At present, mesenchymal stem cells (MSCs) have been isolated from the bone marrow and umbilical cords of giant pandas. However, the types and quantities of preserved cell resources were rare and limited, and none of MSCs was derived from female reproductive organs. Here, we first isolated MSCs from the endometrium of giant panda. These cells showed fibroblast morphology and expressed Sox2, Klf4, Thy1, CD73, CD105, CD44, CD49f, and CD105. Endometrium mesenchymal stem cells (eMSCs) of giant panda could induce differentiation into three germ layers in vitro. RNA-seq analysis showed that 833 genes were upregulated and 716 genes were downregulated in eMSCs compared with skin fibroblast cells. The results of GO and the KEGG analysis of differentially expressed genes (DEGs) were mainly focused on transporter activity, signal transducer activity, pathways regulating pluripotency of stem cells, MAPK signaling pathway, and PI3K-Akt signaling pathway. The genes PLCG2, FRK, JAK3, LYN, PIK3CB, JAK2, CBLB, and MET were identified as hub genes by PPI network analysis. In addition, the exosomes of eMSCs were also isolated and identified. The average diameter of exosomes was 74.26 ± 13.75 nm and highly expressed TSG101 and CD9 but did not express CALNEXIN. A total of 277 miRNAs were detected in the exosomes; the highest expression of miRNA was the has-miR-21-5p. A total of 14461 target genes of the whole miRNAs were predicted and proceeded with functional analysis. In conclusion, we successfully isolated and characterized the giant panda eMSCs and their exosomes, and analyzed their functions through bioinformatics techniques. It not only enriched the conservation types of giant panda cell resources and promoted the protection of genetic diversity, but also laid a foundation for the application of eMSCs and exosomes in the disease treatment of giant pandas.


Assuntos
Exossomos , Células-Tronco Mesenquimais , MicroRNAs , Ursidae , Feminino , Animais , Ursidae/genética , Exossomos/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/genética , Endométrio/metabolismo
9.
Int J Parasitol Parasites Wildl ; 18: 287-291, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35873088

RESUMO

Toxoplasma gondii is a worldwide-distributed zoonotic protozoan parasite which causes toxoplasmosis and has a significant effect on public health. In the giant panda (Ailuropoda melanoleuca), toxoplasmosis can cause asymptomatic infections, reproductive disorder and even death, which poses a serious threat to the conservation of this rare protected species. Therefore, serological investigation of T. gondii is essential to understanding its risk to giant pandas, however, there are no specific testing kits for giant pandas. Previous research has used MAT as the reference method for screening T. gondii, to investigate this further, this study focused on the agreement comparing of MAT with ELISA and IHA tests for detecting T. gondii antibodies in 100 blood samples from 55 captive giant pandas in Chengdu, China. The results showed 87.0%, 87.0%, 84.0%, samples were sero-positive for T. gondii using ELISA (kits a, b, c), respectively, while MAT and IHA tests were 84.0% and 9.0% sero-positive, respectively. There was no significant difference between MAT and the three ELISA kits and these two methods had substantial agreement (0.61 < Ò› ≤ 0.80). Meanwhile, there was a significant difference (P < 0.001) between MAT and IHA, and these two methods had only a slight agreement (қ ≤ 0.20). The relative sensitivity of the ELISA (kits a, b, c) were 89.0%, 91.5% and 95.1%, and the specificity were 86.7%, 80.0% and 80.0%, respectively, which showed these three ELISA kits all had great accuracy. It is suggested that MAT is the recommended test method for primary screening T. gondii in giant pandas and then verified by ELISA.

10.
Genomics Proteomics Bioinformatics ; 20(2): 366-381, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-34487863

RESUMO

Baylisascaris schroederi, a roundworm (ascaridoid) parasite specific to the bamboo-feeding giant panda (Ailuropoda melanoleuca), represents a leading cause of mortality in wild giant panda populations. Here, we present a 293-megabase chromosome-level genome assembly of B. schroederi to infer its biology, including host adaptations. Comparative genomics revealed an evolutionary trajectory accompanied by host-shift events in ascaridoid parasite lineages after host separations, suggesting their potential for transmission and rapid adaptation to new hosts. Genomic and anatomical lines of evidence, including expansion and positive selection of genes related to the cuticle and basal metabolisms, indicate that B. schroederi undergoes specific adaptations to survive in the sharp-edged bamboo-enriched gut of giant pandas by structurally increasing its cuticle thickness and efficiently utilizing host nutrients through gut parasitism. Additionally, we characterized the secretome of B. schroederi and predicted potential drug and vaccine targets for new control strategies. Overall, this genome resource provides new insights into the host adaptation of B. schroederi to the giant panda as well as the host-shift events in ascaridoid parasite lineages. Our findings on the unique biology of B. schroederi will also aid in the development of prevention and treatment measures to protect giant panda populations from roundworm parasitism.


Assuntos
Ascaridoidea , Parasitos , Ursidae , Animais , Ursidae/genética , Ursidae/parasitologia , Ascaridoidea/genética
11.
Front Cell Infect Microbiol ; 12: 1071988, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36519136

RESUMO

Introduction: Toxoplasma gondii, a globally zoonotic protozoan parasite, infects most warm-blooded animals including the giant panda, and poses a serious threat to the giant panda conservation. However, the seroprevalence and the risk factors for toxoplasmosis in giant pandas are unknown. Here we aimed to determine the seroprevalence of T. gondii in the captive population of giant pandas and analyze the factors associated with the increased risk of infection. Methods: A total of 203 serum samples were collected from 157 (95 females and 62 males) captive giant pandas from 2007 to 2022, antibodies against T. gondii were screened using commercial ELISA and MAT kits. Results: The results showed 56 (35.67%) giant pandas were seropositive, age and transfer history between institutions were identifified as risk factors for T. gondii infection. It is suggested that age-related seroprevalence was the main factor, and housing multiple species in the same environment may increase the chance of cross-infection of T. gondii. Discussion: This study can provide research data for developing policies for the prevention and control of T. gondii and protecting the health of captive giant pandas and other wildlife.


Assuntos
Toxoplasma , Toxoplasmose , Ursidae , Animais , Masculino , Feminino , Estudos Soroepidemiológicos , Anticorpos Antiprotozoários , Fatores de Risco
12.
Front Cell Infect Microbiol ; 11: 820144, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-35198456

RESUMO

A feline panleukopenia virus (FPV), Giant panda/CD/2018, was isolated from a captive giant panda with mild diarrhea in 2018 in Chengdu, China, and further identified via indirect immunofluorescence assay (IFA), transmission electron microscopy (TEM) observation, and genetic analysis. Phylogenetic analysis based on the complete VP2 nucleotide sequences showed that it shared high homology with Chinese FPV isolates and grouped within FPV cluster 1. One unique substitution Gly(G)299Glu(E) in the capsid protein VP2 was first identified with Giant panda/CD/2018. The presence of the G299E substitution is notable as it is located on the top region of the interconnecting surface loop 3, which may be involved in controlling the host range and antigenicity of FPV. These findings first demonstrate that FPV with natural point mutation G299E in the VP2 gene is prevalent in giant panda and suggest that etiological surveillance and vaccination among all giant pandas are urgently needed to protect this endangered species against FPV infection.


Assuntos
Vírus da Panleucopenia Felina , Infecções por Parvoviridae , Ursidae , Animais , Animais de Zoológico/virologia , Proteínas do Capsídeo/genética , China/epidemiologia , Diarreia/veterinária , Diarreia/virologia , Vírus da Panleucopenia Felina/genética , Infecções por Parvoviridae/veterinária , Filogenia , Ursidae/virologia
13.
Mol Med Rep ; 21(5): 2260-2266, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32186752

RESUMO

The aim of the present study was to investigate the effect of microRNAs (miRNAs) on the expression level of core1ß3­galactosyltransferase­specific molecular chaperone (COSMC) in immunoglobulin A nephropathy (IgAN). miRNA expression levels were determined in pediatric patients with IgAN (IgAN group), in patients with other renal diseases (control group) and healthy pediatrics (control group). The target miRNAs of COSMC were investigated in the present study. Western blot analysis was performed to examine the effects of miRNAs on COSMC expression levels. In addition, galactose­deficient IgA1 (Gd­IgA1) expression levels were detected following the addition of miRNA­196b. The present results suggested that the expression levels of 205 miRNAs significantly differed between the IgAN and healthy control groups. The present results also suggested that miRNA­196b and miRNA­33a­3p targeted COSMC, and that miRNA­196b expression in B lymphocytes was significantly higher in the IgAN group compared with the control group (P<0.0001). However, COSMC expression level was significantly downregulated in isolated B lymphocytes transfected with miRNA­196b mimics, but Gd­IgA1 expression levels were increased. Therefore, miRNA­196b may play a role in the formation of Gd­IgA1 and IgAN pathogenesis via COSMC regulation.


Assuntos
Galactosiltransferases/metabolismo , Glomerulonefrite por IGA/metabolismo , MicroRNAs/metabolismo , Chaperonas Moleculares/metabolismo , Linfócitos B/metabolismo , Criança , Regulação para Baixo , Feminino , Galactosiltransferases/genética , Humanos , Imunoglobulina A , Masculino , MicroRNAs/genética , Pediatria
14.
Am J Med Sci ; 359(3): 168-176, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32089158

RESUMO

BACKGROUND: Intestinal-barrier damage plays an important pathogenic role in immunoglobulin A nephropathy (IgAN). In this study, we explored the characteristics of the intestinal barrier in rats with IgAN. MATERIALS AND METHODS: We randomly divided 17 Sprague Dawley (SD) male rats into a normal control group (NC; n = 9) and an IgAN model group (n = 8). Feces in the distal ileum were taken for intestinal-microbiota 16sDNA sequencing. We also took a segment of terminal ileum to analyze intestinal morphology and to detect mRNA and protein expression of the tight-junction proteins zonula occludens-1 (ZO-1) and occludin (OCLN), as well as of mucin 2 (MUC2). We then measured levels of serum diamine oxidase (DAO) and D-lactic acid (D-LA), the biomarkers of intestinal permeability. RESULTS: Compared with the NC group, mRNA expression levels of ZO-1 (t = 4.216, P = 0.0007), OCLN (t = 2.413, P = 0.029) and MUC2 (t = 0.859, P < 0.0001) were significantly decreased in the IgAN model group. Protein expression of ZO-1 (t = 7.349, P < 0.0001) and OCLN (t = 6.367, P < 0.0001) was also decreased in the IgAN model group. Conversely, serum DAO (t = 3.758, P = 0.0024) and D-LA (t = 2.246, P = 0.0427) levels increased in this group. At the genus level, the relative abundance of Ruminococcus2 (P = 0.0086) was increased in the IgAN model group. CONCLUSIONS: Decreased expression of ZO-1, OCLN and MUC2, plus intestinal-microbiota dysbiosis, are associated with intestinal-barrier damage in IgAN rats.


Assuntos
Glomerulonefrite por IGA/fisiopatologia , Mucosa Intestinal/fisiologia , Intestinos/fisiopatologia , Junções Íntimas/fisiologia , Animais , Masculino , Permeabilidade , Distribuição Aleatória , Ratos , Ratos Sprague-Dawley
15.
Vet Parasitol ; 160(1-2): 175-9, 2009 Mar 09.
Artigo em Inglês | MEDLINE | ID: mdl-19046806

RESUMO

In this paper, mtDNA gene cytochrome coxidase subunit I (COI) and small subunit ribosomal RNA (12S rDNA) were used to examine the phylogenetic position of Dirofilaria immitis from dogs and red pandas in the evolutionary tree of filarial. Different approaches, including minimal evolution (ME) and maximum parsimony (MP) from distance matrix and character state, were used to evaluate the evolutional relation between Dirofilaria spp. and other species included in the family Onchocercidae. Intra-specific variation was found in COI but not in 12S rDNA. D. immitis and D. repens appear to be sister species.


Assuntos
Dirofilaria immitis/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Filogenia , RNA Ribossômico/genética , Animais , China/epidemiologia , Dirofilariose/epidemiologia , Dirofilariose/parasitologia , Cães , Complexo IV da Cadeia de Transporte de Elétrons/metabolismo , Regulação da Expressão Gênica/fisiologia , RNA Ribossômico/metabolismo , Ursidae/parasitologia
16.
PLoS One ; 13(1): e0191991, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29385201

RESUMO

Surfactin secreted by Bacillus subtilis can confer strong, diverse antipathogenic effects, thereby benefitting the host. Carbon source is an important factor for surfactin production. However, the mechanism that bacteria utilize cellulose, the most abundant substance in the intestines of herbivores, to produce surfactin remains unclear. Here, we used B. subtilis HH2, isolated from the feces of a giant panda, as a model to determine changes in surfactin expression in the presence of different concentrations of cellulose by quantitative polymerase chain reaction and high-performance liquid chromatography. We further investigated the antimicrobial effects of surfactin against three common intestinal pathogens (Escherichia coli, Staphylococcus aureus, and Salmonella enterica) and its resistance to high temperature (60-121°C), pH (1-12), trypsin (100-300 µg/mL, pH 8), and pepsin (100-300 µg/mL, pH 2). The results showed that the surfactin expressed lowest in bacteria cultured in the presence of 1% glucose medium as the carbon source, whereas increased in an appropriate cellulose concentration (0.67% glucose and 0.33% cellulose). The surfactin could inhibit E. coli and Staphylococcus aureus, but did not affect efficiently for Salmonella enterica. The antibacterial ability of surfactin did not differ according to temperature (60-100°C), pH (2-11), trypsin (100-300 µg/mL), and pepsin (100-300 µg/mL; P > 0.05), but decreased significantly at extreme environments (121°C, pH 1 or 12; P < 0.05) compared with that in the control group (37°C, pH = 7, without any protease). In conclusion, our findings indicated that B. subtilis HH2 could increase surfactin expression in an appropriate cellulose environment and thus provide benefits to improve the intestinal health of herbivores.


Assuntos
Antibacterianos/metabolismo , Bacillus subtilis/metabolismo , Celulose/metabolismo , Lipopeptídeos/metabolismo , Animais , Antibacterianos/farmacologia , Meios de Cultura , Lipopeptídeos/farmacologia , Ursidae
17.
Parasit Vectors ; 10(1): 499, 2017 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-29047380

RESUMO

BACKGROUND: Most studies on Enterocytozoon bieneusi are conducted based on the internal transcribed spacer (ITS) region of the rRNA gene, whereas some have examined E. bieneusi population structures. Currently, the population genetics of this pathogen in giant panda remains unknown. The objective of this study was to determine the E. bieneusi population in captive giant pandas in China. RESULTS: We examined 69 E. bieneusi-positive specimens from captive giant pandas in China using five loci (ITS, MS1, MS3, MS4 and MS7) to infer E. bieneusi population genetics. For multilocus genotype (MLG) analysis of E. bieneusi-positive isolates, the MS1, MS3, MS4, and MS7 microsatellite and minisatellite loci were amplified and sequenced in 48, 45, 50 and 47 specimens, respectively, generating ten, eight, nine and five types. We successfully amplified 36 specimens and sequenced all five loci, forming 24 MLGs. Multilocus sequence analysis revealed a strong and significant linkage disequilibrium (LD), indicating a clonal population. This result was further supported by measurements of pairwise intergenic LD and a standardized index of association (I SA) from allelic profile data. The analysis in STRUCTURE suggested three subpopulations in E. bieneusi, further confirmed using right's fixation index (F ST). Subpopulations 1 and 2 exhibited an epidemic structure, whereas subpopulation 3 had a clonal structure. CONCLUSIONS: Our results describe E. bieneusi population genetics in giant pandas for the first time, improving the current understanding E. bieneusi epidemiology in the studied region. These data also benefit future studies exploring potential transmission risks from pandas to other animals, including humans.


Assuntos
Enterocytozoon/isolamento & purificação , Genética Populacional , Microsporidiose/microbiologia , Ursidae/microbiologia , Animais , Técnicas de Tipagem Bacteriana/veterinária , Enterocytozoon/genética , Repetições de Microssatélites/genética , Tipagem de Sequências Multilocus/veterinária
18.
Sci Rep ; 7: 40749, 2017 01 16.
Artigo em Inglês | MEDLINE | ID: mdl-28091600

RESUMO

Luteinizing hormone (LH) is one of the main pituitary hormones that regulate ovulation, however its role has not been studied in giant panda. In this study, we developed an ELISA method for the detection of panda urinary LH. We analyzed urinary hormones of 24 female pandas during 36 breeding periods, we found females could easily be impregnated if the first mating occurred within 10 hours after LH peak. We also found the patterns of the ratios of urinary LH and progestagen in pandas that bred and successfully gave birth were significantly different from those that bred but failed to give birth. These data was the first to provide the urinary LH profiles during the estrous and gestational periods in pandas, and demonstrated that the appearance of the urinary LH peak indicated the timing of ovulation. The LH detection together with estrogen analysis makes the window for successful mating narrower than previously reported. Moreover, detection of urinary LH and progestagen can be used to discriminate between pregnancies and pseudopregnancies/miscarriages in the species. Thus, our findings suggest that LH not only plays a critical role in regulating ovulation but also plays an important role in maintaining pregnancy in the giant panda.


Assuntos
Estrogênios/urina , Ciclo Estral/urina , Hormônio Luteinizante/urina , Progestinas/urina , Ursidae/fisiologia , Ursidae/urina , Animais , Biomarcadores , Ensaio de Imunoadsorção Enzimática , Feminino , Gravidez , Reprodutibilidade dos Testes , Fatores de Tempo
19.
PLoS One ; 12(7): e0182094, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28750056

RESUMO

Baylisascaris schroederi is a common parasite of captive giant pandas. The diagnosis of this ascariasis is normally carried out by a sedimentation-floatation method or PCR to detect eggs in feces, but neither method is suitable for early diagnosis. Fatty acid-binding protein (FABP) and galectin (GAL) exist in various animals and participate in important biology of parasites. Because of their good immunogenicity, they are seen as potential antigens for the diagnosis of parasitic diseases. In this study, we cloned and expressed recombinant FABP and GAL from B. schroederi (rBs-FABP and rBs-GAL) and developed indirect enzyme-linked immunosorbent assays (ELISAs) to evaluate their potential for diagnosing ascariasis in giant pandas. Immunolocalization showed that Bs-FABP and Bs-GAL were widely distributed in adult worms. The ELISA based on rBs-FABP showed sensitivity of 95.8% (23/24) and specificity of 100% (12/12), and that based on rBs-GAL had sensitivity of 91.7% (22/24) and specificity of 100% (12/12).


Assuntos
Ascaridoidea/metabolismo , Proteínas de Ligação a Ácido Graxo/metabolismo , Galectinas/metabolismo , Células Procarióticas/metabolismo , Testes Sorológicos/métodos , Sequência de Aminoácidos , Animais , Western Blotting , Clonagem Molecular , Ensaio de Imunoadsorção Enzimática , Proteínas de Ligação a Ácido Graxo/química , Feminino , Galectinas/química , Immunoblotting , Masculino , Camundongos Endogâmicos ICR , Filogenia , Coelhos , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Sensibilidade e Especificidade , Alinhamento de Sequência , Análise de Sequência de DNA , Ursidae/parasitologia
20.
Parasite ; 24: 1, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28098070

RESUMO

The present review discusses the findings of cryptosporidiosis research conducted in cattle in China and highlights the currently available information on Cryptosporidium epidemiology, genetic diversity, and distribution in China, which is critical to understanding the economic and public health importance of cryptosporidiosis transmission in cattle. To date, 10 Cryptosporidium species have been detected in cattle in China, with an overall infection rate of 11.9%. The highest rate of infection (19.5%) was observed in preweaned calves, followed by that in juveniles (10.69%), postweaned juveniles (9.0%), and adult cattle (4.94%). The dominant species were C. parvum in preweaned calves and C. andersoni in postweaned, juvenile, and adult cattle. Zoonotic Cryptosporidium species (C. parvum and C. hominis) were found in cattle, indicating the possibility of transmission between humans and cattle. Different cattle breeds had significant differences in the prevalence rate and species of Cryptosporidium. This review demonstrates an age-associated, breed-associated, and geographic-related occurrence of Cryptosporidium and provides references for further understanding of the epidemiological characteristics, and for preventing and controlling the disease.


Assuntos
Doenças dos Bovinos/epidemiologia , Criptosporidiose/epidemiologia , Distribuição por Idade , Animais , Bovinos , Doenças dos Bovinos/economia , Doenças dos Bovinos/parasitologia , Doenças dos Bovinos/prevenção & controle , China/epidemiologia , Criptosporidiose/economia , Criptosporidiose/prevenção & controle , Criptosporidiose/transmissão , Cryptosporidium/classificação , Cryptosporidium/genética , Variação Genética , Prevalência
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