scholarly journals Characterization and phylogenetic analysis of multidrug-resistant protein-encoding genes in Trypanosoma evansi isolated from buffaloes in Ngawi district, Indonesia

2019 ◽  
Vol 12 (10) ◽  
pp. 1573-1577
Author(s):  
Mohammad Mirza Nuryady ◽  
Rini Widayanti ◽  
Raden Wisnu Nurcahyo ◽  
Brilyantika Fadjrinatha ◽  
Ahmad Z. S. Fahrurrozi
Keyword(s):  
Drug Resistance ◽  
Phylogenetic Analysis ◽  
Trypanosoma Brucei ◽  
Polymerase Chain Reaction Amplification ◽  
Point Mutations ◽  
Multidrug Resistant ◽  
Trypanosoma Evansi ◽  
Diminazene Aceturate ◽  
Blood Samples ◽  
First Case

Background and Aim: Excessive use of trypanocidal drugs can lead to cases of drug resistance. Multiple cases of resistance have been widely reported for drugs such as isometamidium chloride and diminazene aceturate. These cases deserve serious attention, especially in Indonesia, where the first case was recorded and where the molecular basis of trypanocidal drug resistance has never been evaluated. This study aimed to analyze the multidrug resistance protein (MRP) gene in Trypanosoma evansi isolates, sampled from Indonesia, by focusing on the phylogenetic relationship between these isolates and other Trypanosoma spp. Materials and Methods: A total of 88 blood samples were drawn from buffaloes in the Ngawi district, Indonesia. Animals infected with T. evansi were detected through the microhematocrit technique and Giemsa blood smear methods. Positive blood samples were used to inoculate in male mice (Mus musculus BALB-C strain) as an animal model for culturing the T. evansi. The genomic DNA of the blood taken from the T. evansi-infected mice was used for polymerase chain reaction amplification, sequencing, and phylogenetic analysis. Results: Two genes were analyzed; the first gene detected for T. evansi corresponded to Trypanosoma brucei with a homology of 99% and the second gene to Trypanosoma brucei gambiense, with a homology of 100%. These two genes of the MRP from T. evansi showed clear similarity to the MRPE and MRPA genes of the T. brucei ssp. Conclusion: The MRP gene is conserved on the subspecies level of T. brucei. Only few point mutations were found between various sequences, which mean that the proteins have the same structure. This is important to treat the parasite with the appropriate drugs in the future.

scholarly journals Evaluasi Pemberian Obat Diminazene Aceturate Secara In Vivo Pada Mencit (Mus musculus) yang Diinfeksi Isolat Trypanosoma evansi

2021 ◽  
Vol 39 (2) ◽  
pp. 185
Author(s):  
Reza Yesica ◽  
Bambang Sutrisno ◽  
Wisnu Nurcahyo
Keyword(s):  
Drug Resistance ◽  
Test Dose ◽  
Trypanosoma Evansi ◽  
Buffy Coat ◽  
Diminazene Aceturate ◽  
In Vivo Test ◽  
Negative Results ◽  
Central Java ◽  
And Control

Abstract Surra's disease is caused by Trypanosoma evansi parasite has been established as one of the strategic infectious animal diseases. Drug resistance in this case is one of the major challenges in handle and control them. The aim of this study is to evaluate the provision drug resistance diminazene aceturate (Tryponil®) on Trypanosoma evansi isolate from Pemalang and Brebes Central Java province with in vivo test in mice. Total 50 mice, BALB / c strain, male, 2 months, body weight ± 30 gram are obtained from LPPT-UGM, adapted for one week. Mice were divided into 10 groups consist of 5 each. Each mouse was infected with Trypanosoma evansi by intraperitonial route. Treatment was given when mice had reached the level of parasitemia 108 – 109 trypanosoma / mL of blood this was predicted 24 hours post-infection (Eisler et al., 2001). The administration of the drug tryapanosidal was done intraperitonial with doses 1mg/kg, 3mg / kg, 5 mg / kg and 7mg / kg. Observation of parasitemia did every 2 times in one week till 60 days of observation. Parasitemia observation was performed using 3 techniques. The first method was native examination used a microscope, if the negative results would be followed by MHCT (Microhaematocrit centrifugation Technique) and BCT (Buffy Coat Technique) according to OIE (2012). Data obtained from the treatment group were the level of parasitemia, the number of deaths and the number of live mice from each test dose. The results are analysed by standard logit or probit. The results of this study showed the effects of the drug Dimianzene aceturate on both isolates varied. On Brebes Isolate was effective at doses of 7 mg / kg BW (100%) and 5mg / kg BW (80%), whereas in the effective dose Pemalang isolate at 3 mg dose / kg BW (80%), 5 and 7 mg / kg BW (100%). While at the lowest dose of 1 mg / kg obtained a level of effectiveness of 0% in both isolates. It could be concluded that both isolates have different pathogens and indicate resistance subpopulation to diminazene aceturate.Keywords : diminazene aceturate, in vivo, resistance, Trypanosoma evansi. 

Azithromycin and Diminazene Aceturate Combination Therapy in Experimental Multidrug-resistant Trypanosoma brucei brucei Infection in Albino Rats

2020 ◽  
Vol 282 ◽  
pp. 109138
Author(s):  
Chukwunonso Francis Obi ◽  
Ikenna Onyema Ezeh ◽  
Michael Ikenna Okpala ◽  
Idika Kalu Idika ◽  
Nnamdi Mbe ◽  
...  
Keyword(s):  
Combination Therapy ◽  
Trypanosoma Brucei ◽  
Multidrug Resistant ◽  
Albino Rats ◽  
Trypanosoma Brucei Brucei ◽  
Diminazene Aceturate

The relationship between duration of infection with Trypanosoma brucei in mice and the efficacy of chemotherapy

Parasitology ◽  
1977 ◽  
Vol 75 (2) ◽  
pp. 143-153 ◽  
Author(s):  
F. W. Jennings ◽  
D. D. Whitelaw ◽  
G. M. Urquhart
Keyword(s):  
Drug Resistance ◽  
Drug Treatment ◽  
Trypanosoma Brucei ◽  
Post Infection ◽  
Drug Resistant ◽  
Diminazene Aceturate ◽  
Duration Of Infection ◽  
Permanent Cure ◽  
Dose Levels ◽  
The Relationship

Relapse of infection after drug treatment of trypanosome infections under conditions precluding re-infection has usually been ascribed to drug resistance on the part of the parasite or to under-dosage of the drug. With Trypanosoma brucei infection in mice we have obtained evidence of another type of relapse. In infections resulting from the inoculation of 1 × 105 trypanosomes, derived from a stabilate T. brucei TREU 667, treatment with diminazene aceturate (Berenil) at 40 mg/kg at either 3 or 7 days after infection elicited a permanent cure. If, however, treatment was delayed later than 14 days after infection, then all the mice relapsed. These relapses generally occurred between 20 and 50 days after treatment, but some mice remained aparasitaemic for up to 60 days. The relapsed infections were apparently not due to the survival of ‘drug-resistant’ trypanosomes, as infections derived from a stabilate isolated from a relapsed Berenil-treated mouse were also permanently cured with Berenil if treated 3 days after infection; however, if treatment was delayed until 21 days post-infection, all the mice relapsed. The cause of relapse was not related to the number of parasites inoculated, as infection resulting from initial inocula of 1 × 105 to 1 × 108 trypanosomes were all cured if treated at 3 days after infection, and all eventually relapsed if treatment was delayed until day 21. This type of relapse phenomenon was not confined to T. brucei TREU 667 but also occurred with 5 other stabi-lates of T. brucei after Berenil treatment. Treatment of T. brucei TREU 667 infections with Ethidium and Prothidium at dose levels of 7.5 and 10 mg/kg respectively was also followed by relapse if treatment was delayed for 3 weeks after infection. The possible causes of relapse under these conditions, and its implications in the study of the natural disease, are discussed.

scholarly journals Genetic Variations Associated with Drug Resistance Markers in Asymptomatic Plasmodium falciparum Infections in Myanmar

Genes ◽  
2019 ◽  
Vol 10 (9) ◽  
pp. 692 ◽  
Author(s):  
Yan Zhao ◽  
Ziling Liu ◽  
Myat Thu Soe ◽  
Lin Wang ◽  
Than Naing Soe ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Plasmodium Falciparum ◽  
Multidrug Resistance ◽  
Point Mutations ◽  
Multidrug Resistant ◽  
Genetic Variations ◽  
Chloroquine Resistance ◽  
Geographically Dispersed ◽  
Resistance Markers ◽  
Asymptomatic Infections

The emergence and spread of drug resistance is a problem hindering malaria elimination in Southeast Asia. In this study, genetic variations in drug resistance markers of Plasmodium falciparum were determined in parasites from asymptomatic populations located in three geographically dispersed townships of Myanmar by PCR and sequencing. Mutations in dihydrofolate reductase (pfdhfr), dihydropteroate synthase (pfdhps), chloroquine resistance transporter (pfcrt), multidrug resistance protein 1 (pfmdr1), multidrug resistance-associated protein 1 (pfmrp1), and Kelch protein 13 (k13) were present in 92.3%, 97.6%, 84.0%, 98.8%, and 68.3% of the parasites, respectively. The pfcrt K76T, pfmdr1 N86Y, pfmdr1 I185K, and pfmrp1 I876V mutations were present in 82.7%, 2.5%, 87.5%, and 59.8% isolates, respectively. The most prevalent haplotypes for pfdhfr, pfdhps, pfcrt and pfmdr1 were 51I/59R/108N/164L, 436A/437G/540E/581A, 74I/75E/76T/220S/271E/326N/356T/371I, and 86N/130E/184Y/185K/1225V, respectively. In addition, 57 isolates had three different point mutations (K191T, F446I, and P574L) and three types of N-terminal insertions (N, NN, NNN) in the k13 gene. In total, 43 distinct haplotypes potentially associated with multidrug resistance were identified. These findings demonstrate a high prevalence of multidrug-resistant P. falciparum in asymptomatic infections from diverse townships in Myanmar, emphasizing the importance of targeting asymptomatic infections to prevent the spread of drug-resistant P. falciparum.

scholarly journals Programmable Base Editing in Mycobacterium tuberculosis Using an Engineered CRISPR RNA-Guided Cytidine Deaminase

2021 ◽  
Vol 3 ◽  
Author(s):  
Xin-Yuan Ding ◽  
Si-Shang Li ◽  
Yi-Man Geng ◽  
Mei-Yi Yan ◽  
Guo-Bao Li ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Mycobacterium Tuberculosis ◽  
Molecular Mechanisms ◽  
Cytidine Deaminase ◽  
Point Mutations ◽  
Treatment Strategies ◽  
Multidrug Resistant ◽  
Resistance Mechanisms ◽  
Bacterial Physiology ◽  
Base Editing

Multidrug-resistant Mycobacterium tuberculosis (Mtb) infection seriously endangers global human health, creating an urgent need for new treatment strategies. Efficient genome editing tools can facilitate identification of key genes and pathways involved in bacterial physiology, pathogenesis, and drug resistance mechanisms, and thus contribute to the development of novel treatments for drug-resistant tuberculosis. Here, we report a two-plasmid system, MtbCBE, used to inactivate genes and introduce point mutations in Mtb. In this system, the assistant plasmid pRecX-NucSE107A expresses RecX and NucSE107A to repress RecA-dependent and NucS-dependent DNA repair systems, and the base editor plasmid pCBE expresses a fusion protein combining cytidine deaminase APOBEC1, Cas9 nickase (nCas9), and uracil DNA glycosylase inhibitor (UGI). Together, the two plasmids enabled efficient G:C to A:T base pair conversion at desired sites in the Mtb genome. The successful development of a base editing system will facilitate elucidation of the molecular mechanisms underlying Mtb pathogenesis and drug resistance and provide critical inspiration for the development of base editing tools in other microbes.

scholarly journals Molecular identification and phylogenetic analysis of Trypanosoma evansi in dromedaries (Camelus dromedarius) from Iran

2021 ◽  
Vol 91 (3) ◽  
pp. 297-305
Author(s):  
Somayeh Bahrami ◽  
◽  
Ali R. Alborzi ◽  
Saeid Rahimi Esfahsalari ◽  
Zahra Ziafati
Keyword(s):  
Risk Factors ◽  
Phylogenetic Analysis ◽  
Phylogenetic Tree ◽  
Molecular Identification ◽  
Blood Smear ◽  
Trypanosoma Evansi ◽  
Blood Samples ◽  
Blood Smear Examination ◽  
The World ◽  
Pcr Method

Surra is of great concern to countries in the world such as Iran, which have a considerable camel population. The present study was aimed at determining the prevalence of Trypanosoma evansi in the camels of Iran. A total of 167 blood samples from farmed camels were examined for the presence of T. evansi infection using parasitological and molecular methods. Blood smear examination revealed 10 (6%) positive samples, while the PCR method 14 (8.4%) found positive samples. Age, sex, and region were not determined as risk factors for T. evansi infection in this study. The phylogenetic tree inferred from VSG gene sequences of T. evansi clearly separated the sequences of this study into two clades, A and B, which reflects the intrasequence heterogeneity among Iranian isolates. The phylogenetic tree showed that Iranian T. evansi strains are members of the T. brucei clade.

A diminazene-resistant strain of Trypanosoma brucei brucei isolated from a dog is cross-resistant to pentamidine in experimentally infected albino rats

Parasitology ◽  
2005 ◽  
Vol 132 (1) ◽  
pp. 127-133 ◽  
Author(s):  
B. M. ANENE ◽  
R. C. EZEOKONKWO ◽  
T. I. MMESIRIONYE ◽  
J. N. A. TETTEY ◽  
J. M. BROCK ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Trypanosoma Brucei ◽  
Active Compound ◽  
Careful Consideration ◽  
Albino Rats ◽  
Trypanosoma Brucei Brucei ◽  
Diminazene Aceturate ◽  
Pentamidine Isethionate ◽  
Relapse Rates

Trypanosomosis is a major cause of mortality for dogs in Nigeria and treatment with diminazene aceturate has steadily become less effective, either as a result of low quality of the locally available diminazene preparations or of drug resistance. To investigate these alternatives, samples of locally obtained drugs were analysed for diminazene aceturate content and a strain of Trypanosoma brucei brucei was isolated from a diminazene-refractory dog in Nsukka, south-eastern Nigeria, and used to infect albino rats. The quality of diminazene aceturate-based preparations was variable, with two preparations containing less than 95% of the stated active compound. Rats infected with T. brucei isolated from the dog were treated 7 and 10 days after infection either with 7 mg/kg diminazene aceturate (intraperitoneally, once) or with 4 mg/kg pentamidine isethionate (intramuscularly, 7 consecutive days). Relapse rates were 100% for both trypanocides in the groups of rat treated 10 days post-infection, and 83% and 50% of rats treated 7 days after infection relapsed to diminazene aceturate and pentamidine isethionate, respectively. Careful consideration of physiological parameters showed that pentamidine was only marginally superior to diminazene aceturate as applied in this study. It was concluded that dogs in Nigeria are infected with genuinely diminazene aceturate-resistant trypanosomes that appear to be cross-resistant to pentamidine isethionate.

scholarly journals Antimicrobial susceptibility, serotype distribution, virulence profile and molecular typing of piliated clinical isolates of pneumococci from east coast, Peninsular Malaysia

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Nurul Diana Dzaraly ◽  
Mohd Nasir Mohd Desa ◽  
AbdulRahman Muthanna ◽  
Siti Norbaya Masri ◽  
Niazlin Mohd Taib ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Antimicrobial Susceptibility ◽  
East Coast ◽  
Tertiary Hospital ◽  
Multidrug Resistant ◽  
Peninsular Malaysia ◽  
Clinical Isolates ◽  
Serotype Distribution ◽  
Antimicrobial Susceptibility Test ◽  
Conjugate Vaccines

AbstractPilus has been recently associated with pneumococcal pathogenesis in humans. The information regarding piliated isolates in Malaysia is scarce, especially in the less developed states on the east coast of Peninsular Malaysia. Therefore, we studied the characteristics of pneumococci, including the piliated isolates, in relation to antimicrobial susceptibility, serotypes, and genotypes at a major tertiary hospital on the east coast of Peninsular Malaysia. A total of 100 clinical isolates collected between September 2017 and December 2019 were subjected to serotyping, antimicrobial susceptibility test, and detection of pneumococcal virulence and pilus genes. Multilocus sequence typing (MLST) and phylogenetic analysis were performed only for piliated strains. The most frequent serotypes were 14 (17%), 6A/B (16%), 23F (12%), 19A (11%), and 19F (11%). The majority of isolates were resistant to erythromycin (42%), tetracycline (37%), and trimethoprim-sulfamethoxazole (24%). Piliated isolates occurred in a proportion of 19%; 47.3% of them were multidrug-resistant (MDR) and a majority had serotype 19F. This study showed ST236 was the most predominant sequence type (ST) among piliated isolates, which was related to PMEN clone Taiwan19F-14 (CC271). In the phylogenetic analysis, the piliated isolates were grouped into three major clades supported with 100% bootstrap values. Most piliated isolates belonged to internationally disseminated clones of S. pneumoniae, but pneumococcal conjugate vaccines (PCVs) have the potential to control them.

scholarly journals PATH-17. NOVEL DUAL HISTONE 3 K27M AND G34R POINT MUTATIONS IN A MIDLINE GLIOMA

2020 ◽  
Vol 22 (Supplement_2) ◽  
pp. ii167-ii167
Author(s):  
Peter Pan ◽  
Tejus Bale ◽  
Alexandra Miller ◽  
Marc Ladanyi ◽  
Marc Rosenblum ◽  
...  
Keyword(s):  
Point Mutations ◽  
Histone 3 ◽  
First Case ◽  
Microvascular Proliferation ◽  
Motor Neuron Facial Nerve ◽  
Methylguanine Methyltransferase ◽  
Mitotic Figures ◽  
In Cis ◽  
Generation Sequencing ◽  
Diffuse Midline Glioma

Abstract BACKGROUND Histone H3 alterations due to mutations on H3F3A and HIST1H3B genes, have in recent years been associated with distinct entities and tumor locations within the context of infiltrative gliomas. H3K27M is associated with a midline location and is included in the WHO 2016 as the diffuse midline glioma H3K27M-mutant, a specific diagnostic entity. H3G34R, thought to be a mutually exclusive alteration, is less common but has been associated with a cerebral hemispheric location. We report the first case to our knowledge with both of these alterations in the same tumor. METHODS Clinical and pathologic records of the patient were reviewed and presented. RESULTS A 39-year-old man presented with acute right face, arm, and leg numbness and mild weakness; examination was notable for right lower motor neuron facial nerve palsy and numbness, along with numbness and subtle slowing of rapid alternating movements in the left arm and leg. A non-enhancing left thalamic mass was identified and stereotactically biopsied. Infiltrative glial neoplasm with moderately-increased cellularity was seen, with ovoid cells, enlarged nuclei, apoptotic bodies, and mitotic figures. No necrosis or microvascular proliferation seen. Immunostain was positive for H3K27M. O6-methylguanine-methyltransferase (MGMT) promoter was not methylated. Next-generation sequencing showed dual in cis H3 point mutations in K27M (HIST1H3B c.83A >T) and G34R (HIST1H3B c.103G >C). Additional alterations were noted in NF1, PIK3CA, ATRX, FGFR3, and NSD1. Isocitrate dehydrogenase (IDH1/2) mutations were not identified. CONCLUSION This case of a young man with a midline glioma is novel for carrying both H3K27M and H3G34R alterations and indicates these alterations are not mutually exclusive. The interaction seen here suggests H3K27M dominance for a midline phenotype.

scholarly journals Analysis of drug resistance and mutation profiles in Mycobacterium tuberculosis isolates in a surveillance site in Beijing, China

2021 ◽  
Vol 49 (1) ◽  
pp. 030006052098493
Author(s):  
Jie Zhang ◽  
Yixuan Ren ◽  
Liping Pan ◽  
Junli Yi ◽  
Tong Guan ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Mycobacterium Tuberculosis ◽  
Drug Testing ◽  
Multidrug Resistant ◽  
High Rate ◽  
Drug Resistant ◽  
Surveillance Site ◽  
Mdr Tb ◽  
Previously Treated Patients ◽  
Beijing China

Objective This study analyzed drug resistance and mutations profiles in Mycobacterium tuberculosis isolates in a surveillance site in Huairou District, Beijing, China. Methods The proportion method was used to assess drug resistance profiles for four first-line and seven second-line anti-tuberculosis (TB) drugs. Molecular line probe assays were used for the rapid detection of resistance to rifampicin (RIF) and isoniazid (INH). Results Among 235 strains of M. tuberculosis, 79 (33.6%) isolates were resistant to one or more drugs. The isolates included 18 monoresistant (7.7%), 19 polyresistant (8.1%), 28 RIF-resistant (11.9%), 24 multidrug-resistant (MDR) (10.2%), 7 pre-extensively drug-resistant (XDR, 3.0%), and 2 XDR strains (0.9%). A higher rate of MDR-TB was detected among previously treated patients than among patients with newly diagnosed TB (34.5% vs. 6.8%). The majority (62.5%) of RIF-resistant isolates exhibited a mutation at S531L in the DNA-dependent RNA polymerase gene. Meanwhile, 62.9% of INH-resistant isolates carried a mutation at S315T1 in the katG gene. Conclusion Our results confirmed the high rate of drug-resistant TB, especially MDR-TB, in Huairou District, Beijing, China. Therefore, detailed drug testing is crucial in the evaluation of MDR-TB treatment.

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