scholarly journals Occurrence and multilocus genotyping of Giardia duodenalis in pets and zoo animals in Shanghai, China

2017 ◽  
Vol 11 (06) ◽  
pp. 479-486 ◽  
Author(s):  
Hua Liu ◽  
Yujuan Shen ◽  
Aiqin Liu ◽  
Jianhai Yin ◽  
Zhongying Yuan ◽  
...  
Keyword(s):  
Giardia Duodenalis ◽  
Pcr Amplification ◽  
Amino Acid Sequences ◽  
Giardia Infection ◽  
Multilocus Genotyping ◽  
High Prevalence ◽  
Amplification Rate ◽  
Zoo Animals ◽  
First Time

Introduction: High prevalence of Giardia infections occurs in humans and animals, partly because of the increasing numbers of pets. We determined the presence and genotypes of G. duodenalis in pets and zoo animals. Methodology: A total of 84 specimens were collected from dogs and cats from a pet hospital, and 54 specimens from a zoo, which included deer, tigers, yaks, and others. All the specimens were examined by microscopy and by polymerase chain reaction (PCR) amplification and subsequent sequencing of glutamate dehydrogenase (gdh), beta-giardin (bg), and triose phosphate isomerase (tpi) genes. Results: Giardia infection was confirmed in 5.95% and 15.48% of animals by microscopy and by PCR, respectively; the detection levels were 13.33% and 26.67% for pets, and 1.85% and 9.26% for zoo animals. Four assemblages were identified: assemblage C in dogs, cats, and a sheep; D in dogs, a wolf, a yak, and a leopard; E in a sheep; and F in a cat and a leopard. PCR gave the highest amplification rate at the gdh locus. Eight, five, and four sequences were novel at the gdh, bg, and tpi loci, respectively. Two tpi sequences of dog-derived assemblage C had 100% homology with amino acid sequences from human-derived isolates. Conclusions: The molecular characterization of G. duodenalis in pets and zoo animals in China is described. Assemblage D was identified in a yak and a leopard for the first time. Multilocus genotyping analysis identified the same tpi gene sequences of assemblage C in dogs and humans, indicating potential zoonotic transmission.

Characterization of a cDNA for Rhesus Monkey Protein C Inhibitor – Evidence for N-Terminal Involvement in Heparin Stimulation

1995 ◽  
Vol 74 (04) ◽  
pp. 1079-1087 ◽  
Author(s):  
Klaus-P Radtke ◽  
José A Fernández ◽  
Bruno O Villoutreix ◽  
Judith S Greengard ◽  
John H Griffin
Keyword(s):  
Rhesus Monkey ◽  
Protein C ◽  
Activated Protein C ◽  
Pcr Amplification ◽  
Amino Acid Sequences ◽  
Heparin Binding ◽  
Reactive Center ◽  
Protein C Inhibitor ◽  
Polymerase Chain

SummarycDNAs for protein C inhibitor (PCI) were cloned from human and rhesus monkey 1 liver RNAs by reverse transcription and polymerase chain reaction (PCR) amplification. Sequencing showed that rhesus monkey and human PCI cDNAs were 93% identical. Predicted amino acid sequences differed at 26 of 387 residues. Pour of these differences (T352M, N359S, R362K, L3631) were in the reactive center loop that is important for inhibitory specificity, and two were in the N-terminal helix (M8T, E13K) that is implicated in glycosaminoglycan binding. PCI in human or rhesus monkey plasma showed comparable inhibitory activity towards human activated protein C in the presence of 10 U/ml heparin. However, maximal acceleration of the inhibition of activated protein C required 5-fold lower heparin concentration for rhesus monkey than for human plasma, consistent with the interpretation that the additional positive charge (E13K) in a putative-heparin binding region increased the affinity for heparin.

scholarly journals Molecular characterization of Prunus necrotic ringspot virus isolated from rose in Brazil

2015 ◽  
Vol 45 (12) ◽  
pp. 2197-2200 ◽  
Author(s):  
Thor Vinícius Martins Fajardo ◽  
Monique Bezerra Nascimento ◽  
Marcelo Eiras ◽  
Osmar Nickel ◽  
Gilvan Pio-Ribeiro
Keyword(s):  
Amino Acid ◽  
Molecular Characterization ◽  
Molecular Analysis ◽  
Nucleotide Sequences ◽  
Amino Acid Sequences ◽  
Causal Agent ◽  
Ringspot Virus ◽  
Prunus Necrotic Ringspot Virus ◽  
First Time

ABSTRACT: There is no molecular characterization of Brazilian isolates of Prunus necrotic ringspot virus (PNRSV), except for those infecting peach. In this research, the causal agent of rose mosaic was determined and the movement (MP) and coat (CP) protein genes of a PNRSV isolate from rose were molecularly characterized for the first time in Brazil. The nucleotide and deduced amino acid sequences of MP and CP complete genes were aligned and compared with other isolates. Molecular analysis of the MP and CP nucleotide sequences of a Brazilian PNRSV isolate from rose and others from this same host showed highest identities of 96.7% and 98.6%, respectively, and Rose-Br isolate was classified in PV32 group.

scholarly journals Enzymic and structural characterization of nepenthesin, a unique member of a novel subfamily of aspartic proteinases

2004 ◽  
Vol 381 (1) ◽  
pp. 295-306 ◽  
Author(s):  
Senarath B. P. ATHAUDA ◽  
Koji MATSUMOTO ◽  
Sanath RAJAPAKSHE ◽  
Masayuki KURIBAYASHI ◽  
Masaki KOJIMA ◽  
...  
Keyword(s):  
Amino Acid ◽  
Computer Modelling ◽  
Structural Characteristics ◽  
Amino Acid Sequences ◽  
Cysteine Residues ◽  
Aspartic Proteinases ◽  
Protein Databases ◽  
Wide Range ◽  
First Time

Carnivorous plants are known to secrete acid proteinases to digest prey, mainly insects, for nitrogen uptake. In the present study, we have purified, for the first time, to homogeneity two acid proteinases (nepenthesins I and II) from the pitcher fluid of Nepenthes distillatoria (a pitcher-plant known locally as badura) and investigated their enzymic and structural characteristics. Both enzymes were optimally active at pH approx. 2.6 towards acid-denatured haemoglobin; the specificity of nepenthesin I towards oxidized insulin B chain appears to be similar, but slightly wider than those of other APs (aspartic proteinases). Among the enzymic properties, however, the most notable is their unusual stability: both enzymes were remarkably stable at or below 50 °C, especially nepenthesin I was extremely stable over a wide range of pH from 3 to 10 for over 30 days. This suggests an evolutionary adaptation of the enzymes to their specific habitat. We have also cloned the cDNAs and deduced the complete amino acid sequences of the precursors of nepenthesins I and II (437 and 438 residues respectively) from the pitcher tissue of N. gracilis. Although the corresponding mature enzymes (each 359 residues) are homologous with ordinary pepsin-type APs, both enzymes had a high content of cysteine residues (12 residues/molecule), which are assumed to form six unique disulphide bonds as suggested by computer modelling and are supposed to contribute towards the remarkable stability of nepenthesins. Moreover, the amino acid sequence identity of nepenthesins with ordinary APs, including plant vacuolar APs, is remarkably low (approx. 20%), and phylogenetic comparison shows that nepenthesins are distantly related to them to form a novel subfamily of APs with a high content of cysteine residues and a characteristic insertion, named ‘the nepenthesin-type AP-specific insertion’, that includes a large number of novel, orthologous plant APs emerging in the gene/protein databases.

scholarly journals Detection of Human Genotype “B” Giardia lamblia in Ghanaian Cattle from Frafraha in Adentan Municipality of Ghana

2020 ◽  
Vol 61 (1) ◽  
pp. 96-100
Author(s):  
G. T. Mensah ◽  
C. A. Narh ◽  
C. A. Brown ◽  
P. F. Ayeh-Kumi ◽  
I. O. Frempong
Keyword(s):  
Giardia Lamblia ◽  
Intestinal Parasite ◽  
Giardia Duodenalis ◽  
Pcr Amplification ◽  
Faecal Samples ◽  
Wide Range ◽  
Close Proximity ◽  
First Time ◽  
Animal Research Institute ◽  
Human Specific

Giardia duodenalis is a common intestinal parasite in humans, a wide range of domesticated and wild animals. There are human and animal specific, as well as zoonotic pathogenic gen­otypes. It is not clear whether livestock in close proximity to humans could be infected with human specific genotypes, and vice versa. In this study, Giardia-positive faecal samples were collected from both humans (n = 4) (from Maamobi Polyclinic in the Ayawaso Sub- Metro) and calves (n = 8) (from Animal Research Institute Farms, Adentan Municipality), in Ghana. Nested PCR amplification using Giardia-specific, Glutamate dehydrogenase (GDH) genes and Tri­osephosphate isomerase (TPI) gene primers for human and animal faecal samples respectively was carried out. Results showed that 100% of the calves carried the TPI-B genotype, which is a common pathogenic genotype in humans. This report is based on the unusual results obtained as Giardia duodenalis genotype B is known to occur in humans but is being observed in calves for the first time. This suggests that calves in close proximity to humans could be reservoirs and sources of human Giardia infections.

scholarly journals Intestinal Parasites of Buffalo Calves from Romania: Molecular Characterization of Cryptoporidium spp. and Giardia Duodenalis, and the first Report of Eimeria Bareillyi

2021 ◽  
Author(s):  
Diana Ancuța Bărburaș ◽  
Vasile Cozma ◽  
Angela Monica Ionică ◽  
Ibrahim Abbas ◽  
Remus Bărburaș ◽  
...  
Keyword(s):  
Intestinal Parasites ◽  
Giardia Duodenalis ◽  
Gastrointestinal Parasites ◽  
Parasitic Infections ◽  
Buffalo Calves ◽  
Economic Resource ◽  
Toxocara Vitulorum ◽  
Staining Methods ◽  
First Time

Abstract Buffaloes represent an important economic resource for several regions of the world including Romania; however, no reports on parasitic infections in buffaloes from Romania are available. In the present study, we examined for the gastrointestinal parasites 104 fecal samples bimonthly collected from 38 buffalo calves (2–11 weeks old) from household rearing systems in Romania. All samples were tested using the saturated salt flotation, McMaster and modified Ziehl-Nielsen staining methods. PCR coupled with isolates sequencing methods were used to identify the Giardia duodenalis assemblages and Cryptosporidium species. Overall, 33 out of 38 examined buffalo calves were infected with different gastrointestinal parasites; 16 had single infections and 17 had mixed infections with 2 or 3 parasites. Eimeria species (32/38; 84.2%) was the most prevalent parasite; 8 species were identified according to the oocyst morphology including the pathogenic E. bareillyi which detected for the first time in buffaloes from Romania. Toxocara vitulorum (11/38; 36.8%) and Strongyloides papillosus (6/38; 15.8%) were also detected. Cryptosporidium spp. were found in 4 (10.5%) buffalo calves; 2 of them were molecularly identified as C. ryanae and another one was clustered in the same clade with C. ryanae, C. bovis, and C. xiaoi. Giardia duodenalis assemblage E was also molecularly detected in a single (2.6%) buffalo calf. The presence of other buffaloes in the same barn was identified as a risk factor for infection with T. vitulorum. Our results indicate extensive parasitic infections in buffalo calves from Northwestern Romania and underline the necessity of prophylactic treatments for T. vitulorum and E. bareillyi.

scholarly journals Molecular Detection and Characterization of Goat Isolate ofTaenia hydatigenain Turkey

2012 ◽  
Vol 2012 ◽  
pp. 1-4 ◽  
Author(s):  
Armagan Erdem Utuk ◽  
Fatma Cigdem Piskin
Keyword(s):  
Ribosomal Rna ◽  
Molecular Detection ◽  
Sequence Data ◽  
Pcr Amplification ◽  
Small Subunit ◽  
Molecular Tools ◽  
Taenia Hydatigena ◽  
Nucleotide Sequence Data ◽  
First Time

The aim of this study was to provide molecular detection and characterization of the goat isolate ofTaenia hydatigenafrom Ankara province of Turkey. For this purpose, PCR amplification of small subunit ribosomal RNA (rrnS) and partial sequencing of mitochondrial cytochrome c oxidase subunit 1 (mt-CO1) genes were performed in a one-month-old dead goat. According to rrnS-PCR results, parasites were identified asTaeniaspp., and partial sequence of mt-CO1 gene was corresponding toT. hydatigena. At the end of the study, we concluded that molecular tools can be used to define species of parasites in cases where the key morphologic features cannot be detected. Nucleotide sequence data of Turkish goat isolate ofT. hydatigenawas submitted to GenBank for other researchers interested in this subject. By this study, molecular detection and characterization ofT. hydatigenawas done for the first time in Turkey.

scholarly journals Detection and partial molecular characterization of Grapevine fleck virus, Grapevine virus D, Grapevine leafroll-associated virus -5 and -6 infecting grapevines in Brazil

2012 ◽  
Vol 42 (12) ◽  
pp. 2127-2130 ◽  
Author(s):  
Thor Vinícius Martins Fajardo ◽  
Marcelo Eiras ◽  
Osmar Nickel ◽  
Carla Rosa Dubiela ◽  
Eliezer Rodrigues de Souto
Keyword(s):  
Coat Protein ◽  
Molecular Characterization ◽  
Rna Binding ◽  
Rna Binding Protein ◽  
Amino Acid Sequences ◽  
Viral Diseases ◽  
Grapevine Virus ◽  
Available Information ◽  
First Time

Grapevine fleck, rugose wood and leafroll are three grapevine viral diseases whose causal agents (or associated viruses) respectively are Grapevine fleck virus (GFkV), Grapevine virus D (GVD) and Grapevine leafroll-associated virus 5 and 6 (GLRaV-5 and -6). The objective of this work was to perform a partial molecular characterization of local isolates of these four viral species that infect grapevines. The nucleotide and deduced amino acid sequences of complete genes of the coat protein (CP) (of GFkV), the CP and the RNA binding protein (of GVD), the CP and the partial hHSP70 gene (of GLRaV-5) and the partial hHSP70 gene (of GLRaV-6) were aligned and compared in silico with other isolates. These data extend the available information about Brazilian isolates of GFkV, GLRaV-5 and -6, and reports for the first time the GVD occurrence in Brazil.

scholarly journals Prevalence and Molecular Characterization of Giardia duodenalis in Calves in Turkey

2017 ◽  
Vol 45 (1) ◽  
pp. 6
Author(s):  
Mehmet Gultekin ◽  
Kerem Ural ◽  
Nuram Aysul ◽  
Adnam Ayan ◽  
Canberk Balikci ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Nested Pcr ◽  
Giardia Duodenalis ◽  
Pcr Assay ◽  
Giardia Infection ◽  
Western Turkey ◽  
Aegean Region ◽  
Animal Populations ◽  
Faecal Samples

Background: Giardia duodenalis (G. duodenalis) is an ubiquitous, flagellated intestinal protozoan with major public health significance worldwide. Limited data are available on the epidemiology of G. duodenalis in dairy cattle from Turkey. Determining the zoonotic potential of the Giardia infection requires molecular characterization. The aim of the present study was to investigate the prevalence and to molecularly characterize G. duodenalis in calves less than three months of age in Aydin, Aegean region of Turkey.Materials, Methods & Results: The study was conducted on different dairy farms in the south-western part of the Turkey, Aegean Region, Aydin. A total of 198 Holstein Friesian calves less than three months of age, of both sexes were enrolled into the study. Faecal samples from each calf were collected manually from the rectum using a disposable latex glove. The consistency of collected samples was recorded as diarrhoeic or non-diarrhoeic.  Diagnosis of G. duodenalis infection was made microscopically by detection of cysts in the faecal samples. One hundred and sixteen (58.5%) of the 198 faecal samples were diarrheic. Giardia cysts were found in 27 (23.28%) of the diarrheic samples and in 8 (9.76%) of nondiarrheic samples (P < 0.05). The overall prevalence of giardiosis in calves was determined as 17.67%. The prevalence of Giardia genotypes was identified by DNA sequence analysis of the beta-giardin gene for every PCR positive sample. The beta-giardin nested PCR assay was revealed assemblage A and sub-genotype A3 was detected in all of 35 samples (100%).Discussion: The highest prevalence of Giardia infection in calves is reported at the age between 1 and 6 months, and the prevalence shows decreased rate from the age of 6 months. The present study was conducted in Aydin, a province of south-western Turkey in the Aegean Region, and the overall prevalence from a total of 198 dairy calves was 17.67%. The prevalence rate in calves with diarrhoea was higher and reached up to 23.28%, whereas it was 9.76% in non-diarrhoeic calves. A prevalence study with molecular characterization of G. duodenalis isolates in cattle has not yet been reported from Turkey. Molecular studies have shown that mostly assemblage E predominates in cattle, but recent studies denoted that assemblage A is increasingly being detected and might be more widespread than expected before. In the present study, Giardia positive samples identified with a beta-giardin nested PCR assay. The sub-genotype A3 was identified in all samples. The same sub-genotype was identified in human and dog samples from different countries. Furthermore, sub-genotype A3 was found in humans and dogs from Turkey. In this context, results of the present study suggested an important role of calves as potential reservoirs of human infections in Turkey. In conclusion, epidemiological data revealed that G. duodenalis infection is frequent in calves with diarrhoea in Aydin, Turkey. The presence of the potentially zoonotic sub-genotype A3 and high prevalence of Giardia infection in diarrheic calves indicated the importance of treatment and necessary preventative measures. Further studies in human and animal populations living in this region are warranted regarding the zoonotic epidemiology of Giardia duodenalis.

New insight in cereal starch degradation: identification and structural characterization of four α-amylases in bread wheat

Amylase ◽  
2017 ◽  
Vol 1 (1) ◽  
Author(s):  
Jos C. Mieog ◽  
Štefan Janeček ◽  
Jean-Philippe Ral
Keyword(s):  
Starch Granule ◽  
Amylase Activity ◽  
Amino Acid Sequences ◽  
Starch Degradation ◽  
Apparent Paradox ◽  
Germination Process ◽  
Late Maturity ◽  
First Time ◽  
Comprehensive Study

AbstractGrain α-amylase presents an apparent paradox for the wheat community. Despite the necessity of α-amylase for the seed germination process, high levels of amylase activity in the grain are considered detrimental for grain quality. Wheat α-amylases (EC 3.2.1.1) are endohydrolases belonging to the GH13_6 subfamily, one of the most studied subclasses of glycoside hydrolase (GH) family GH13. However, no comprehensive study had been done so far to describe and catalogue all the wheat α-amylase isoforms, despite compelling information on the involvement of two α-amylases on economically important issues for the international cereal community, namely pre-harvest sprouting and late maturity α-amylase. This study describes for the first time the genomic localization, nucleotide and amino acid sequences, phylogeny and expression profile of all known α-amylases in wheat, including a hitherto unknown fourth isoform here designated as TaAMY4. Isoform profiling strongly suggested α-amylases to be working in partnership to achieve complete degradation of a starch granule, whereas expression profiling revealed a potential involvement of TaAMY4 in the late maturity α-amylase problem.

scholarly journals Cloning and Characterization of Benzoate Catabolic Genes in the Gram-Positive Polychlorinated Biphenyl DegraderRhodococcus sp. Strain RHA1

2001 ◽  
Vol 183 (22) ◽  
pp. 6598-6606 ◽  
Author(s):  
Wataru Kitagawa ◽  
Keisuke Miyauchi ◽  
Eiji Masai ◽  
Masao Fukuda
Keyword(s):  
Polychlorinated Biphenyl ◽  
Pcr Amplification ◽  
Gene Organization ◽  
Amino Acid Sequences ◽  
Bacterial Degradation ◽  
High Concentration ◽  
Catabolic Genes ◽  
Electrophoresis Separation ◽  
Nucleotide Sequence Determination

ABSTRACT Benzoate catabolism is thought to play a key role in aerobic bacterial degradation of biphenyl and polychlorinated biphenyls (PCBs). Benzoate catabolic genes were cloned from a PCB degrader,Rhodococcus sp. strain RHA1, by using PCR amplification and temporal temperature gradient electrophoresis separation. A nucleotide sequence determination revealed that the deduced amino acid sequences encoded by the RHA1 benzoate catabolic genes, benABCDK, exhibit 33 to 65% identity with those of Acinetobacter sp. strain ADP1. The gene organization of the RHA1 benABCDKgenes differs from that of ADP1. The RHA1 benABCDK region was localized on the chromosome, in contrast to the biphenyl catabolic genes, which are located on linear plasmids. Escherichia coli cells containing RHA1 benABCD transformed benzoate to catechol via 2-hydro-1,2-dihydroxybenzoate. They transformed neither 2- nor 4-chlorobenzoates but did transform 3-chlorobenzoate. The RHA1 benA gene was inactivated by insertion of a thiostrepton resistance gene. The resultant mutant strain, RBD169, neither grew on benzoate nor transformed benzoate, and it did not transform 3-chlorobenzoate. It did, however, exhibit diminished growth on biphenyl and growth repression in the presence of a high concentration of biphenyl (13 mM). These results indicate that the cloned benABCD genes could play an essential role not only in benzoate catabolism but also in biphenyl catabolism in RHA1. Six rhodococcal benzoate degraders were found to have homologs of RHA1benABC. In contrast, two rhodococcal strains that cannot transform benzoate were found not to have RHA1 benABChomologs, suggesting that many Rhodococcus strains contain benzoate catabolic genes similar to RHA1 benABC.

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