Levels of Ancylostoma infections and phylogenetic analysis of cox 1 gene of A. ceylanicum in stray cat faecal samples from Guangzhou, China

2015 ◽  
Vol 90 (4) ◽  
pp. 392-397 ◽  
Author(s):  
W. Hu ◽  
X.G. Yu ◽  
S. Wu ◽  
L.P. Tan ◽  
M.R. Song ◽  
...  
Keyword(s):  
South China ◽  
Potential Risk ◽  
Stray Cats ◽  
Faecal Samples ◽  
High Prevalence ◽  
Pcr Rflp ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Stray Cat ◽  
A Minor

AbstractAncylostoma ceylanicum is a common zoonotic nematode. Cats act as natural reservoirs of the hookworm and are involved in transmitting infection to humans, thus posing a potential risk to public health. The prevalence of feline A. ceylanicum in Guangzhou (South China) was surveyed by polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP). In total, 112 faecal samples were examined; 34.8% (39/112) and 43.8% (49/112) samples were positive with hookworms by microscopy and PCR method, respectively. Among them, 40.8% of samples harboured A. ceylanicum. Twelve positive A. ceylanicum samples were selected randomly and used for cox 1 sequence analysis. Sequencing results revealed that they had 97–99% similarity with A. ceylanicumcox 1 gene sequences deposited in GenBank. A phylogenetic tree showed that A. ceylanicum isolates were divided into two groups: one comprising four isolates from Guangzhou (South China), and the other comprising those from Malaysia, Cambodia and Guangzhou. In the latter group, all A. ceylanicum isolates from Guangzhou were clustered into a minor group again. The results indicate that the high prevalence of A. ceylanicum in stray cats in South China poses a potential risk of hookworm transmission from pet cats to humans, and that A. ceylanicum may be a species complex worldwide.

scholarly journals Sarcocystis spp. prevalence in bovine minced meat: a histological and molecular study

2015 ◽  
Vol 4 (2) ◽  
Author(s):  
Serena Meistro ◽  
Simone Peletto ◽  
Marzia Pezzolato ◽  
Katia Varello ◽  
Mario Botta ◽  
...  
Keyword(s):  
Public Health Problem ◽  
Molecular Study ◽  
Chain Reaction ◽  
Sarcocystis Spp ◽  
High Prevalence ◽  
Polymerase Chain ◽  
A Minor ◽  
Minced Meat ◽  
Butcher Shops ◽  
Piedmont Region

Sarcosporidiosis is caused by ingestion of contaminated raw or undercooked bovine meat and, although considered a minor zoonosis, it can represent a threath for immunocompromised people. Aim of this study was to determine the prevalence of <em>Sarcocystis</em> spp. in bovine minced meat intended for raw consumption collected from butcher shops and retail stores in Turin’s province (Piedmont region, Northwest Italy). Twenty-five samples were examined in parallel by histology and polymerase chain reaction (PCR). The prevalence of infestation of <em>Sarcocystis</em> spp. resulted to be 64% [confidence interval (CI) 95% 42-82] and 88% (CI 95% 69-97) respectively by histology and PCR. In detail, the prevalence resulted 80% for <em>S. cruzi</em> (CI 95% 59-93), 68% for <em>S. hominis</em> (CI 95% 46-85) and 4% for <em>S. hirsuta</em> (CI 95% 0.10-20). The high prevalence of <em>S. hominis</em> highlights that sarcosporidiosis may constitute a public health problem in Italy, particularly in regions like Piedmont, that has traditional dishes prepared from raw or undercooked bovine meat.

Molecular identification of zoonotic hookworm species in dog faeces from Tanzania

2018 ◽  
Vol 93 (3) ◽  
pp. 313-318 ◽  
Author(s):  
A. Merino-Tejedor ◽  
P. Nejsum ◽  
E.M. Mkupasi ◽  
M.V. Johansen ◽  
Annette Olsen
Keyword(s):  
Sanger Sequencing ◽  
Molecular Techniques ◽  
Health Concern ◽  
Specific Pcr ◽  
Molecular Approaches ◽  
Faecal Samples ◽  
Pcr Rflp ◽  
Veterinary Importance ◽  
Polymerase Chain ◽  
Species Specific

AbstractThe presence and distribution of various species of canine hookworms in Africa are poorly known. The main objective of this study, therefore, was to identify the hookworm species present in canine faecal samples from Morogoro, Tanzania, using molecular techniques. Faecal samples from 160 local dogs were collected and hookworm positive samples processed to recover larvae for further molecular characterization. DNA was extracted from pools of larvae from individual samples (n = 66), which were analysed subsequently using two different molecular approaches, polymerase chain reaction-linked restriction fragment length polymorphism (PCR-RFLP) and species-specific PCR coupled with Sanger sequencing. The PCR-RFLP technique detected only the presence of the ubiquitousAncylostoma caninumin the 66 samples. However, by species-specific PCR coupled with Sanger sequencing we identified ten samples withA. braziliense, two withUncinaria stenocephalaand five withA. ceylanicum. Thus, all four known species of canine hookworms were identified in Morogoro, Tanzania. To our knowledge this is the first report of the detection of the presence ofU. stenocephalaandA. ceylanicumin Africa using molecular techniques. In addition to their veterinary importance, canine hookworms have zoonotic potential and are of public health concern.

scholarly journals Fasciola hepaticain Some Buffaloes and Cattle by PCR and Microscopy

2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Sultan Ayaz ◽  
Riaz Ullah ◽  
Naser M. AbdEl-Salam ◽  
Sumiara Shams ◽  
Sadaf Niaz
Keyword(s):  
Liver Biopsy ◽  
Economic Losses ◽  
Agarose Gel ◽  
Chain Reaction ◽  
Entire Study ◽  
Microscopy Technique ◽  
Faecal Samples ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Livestock Rearing

Fasciolosis is the burning problem of the livestock rearing community having huge morbidity, mortality, and economic losses to livestock industries in our country Pakistan. The faecal and liver biopsy samplings were examined by polymerase chain reaction (PCR) and microscopy technique during the entire study. A total of 307 samples including 149 samples from Karak and 158 samples from Kohat abattoirs were examined by PCR method and overall prevalence of fasciolosis was 5.86% (18/307), amongst theses 8.05% (12/149) in liver biopsy and 3.79% (6/158) in feacal samples of cattle and Buffaloes were recorded. Similarly the microscopy based detection was 3.58% (11/307) including 4.61% (7/149) in liver biopsy and 2.5% (4/158) in faecal samples accordingly. Furthermore the areawise prevalence of fasciolosis in abattoirs by PCR method was found to be 7.59% (12/158) in Kohat and 4.02% (6/149) in Karak. A 618 pb DNA was amplified in 2% agarose gel electrophoreses. It is concluded from the study that prevalence of fasciolosis was higher in abattoir of district Kohat and PCR was a more sensitive method of diagnosis than microscopy.

scholarly journals Identification of Eutypa lata by PCR-RFLP

Plant Disease ◽  
2004 ◽  
Vol 88 (9) ◽  
pp. 925-929 ◽  
Author(s):  
P. E. Rolshausen ◽  
F. Trouillas ◽  
W. D. Gubler
Keyword(s):  
Universal Primers ◽  
Phaeomoniella Chlamydospora ◽  
Phomopsis Viticola ◽  
Eutypa Lata ◽  
Incorrect Diagnosis ◽  
Pcr Rflp ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Rflp Assay

Eutypa lata is a vascular canker pathogen of woody plants commonly diagnosed by isolating the pathogen from infected tissue. Related fungi from the same family, the Diatrypaceae, also have been found in association with grapevine in Californian vineyards. An in situ polymerase chain reaction (PCR) method has been developed for detection of E. lata in infected wood tissue. However, our results indicate that this method also would amplify other Diatrypaceous fungi, which could potentially lead to an incorrect diagnosis. Therefore, we developed a PCR-restriction fragment length polymorphism (PCR-RFLP) assay. The internal transcribed spacer (ITS)1/5.8S/ITS2 ribosomal DNA region was amplified by PCR using universal primers, and RFLP patterns were determined after digestion with AluI. The restriction profiles obtained served to distinguish E. lata from wood trunk pathogens of grapevine (Phomopsis viticola, Botryodiplodia sp., Phaeoacremonium aleophilum, and Phaeomoniella chlamydospora), Diatrypaceous fungi (Diatrype sp., Diatrypella sp., Eutypella vitis, and Eutypa leptoplaca), and Cryptovalsa sp. found on dead wood of grapevine, and other Eutypa spp. (E.petrakii var. hederae, E. astroidea, E. crustata, and E. lejoplaca), with the exception of E. armeniacae, which we regard as a synonym for E. lata, and E. laevata, which has not been found on grapevine.

scholarly journals katG (SER 315 THR) Gene Mutation in Isoniazid Resistant Mycobacterium Tuberculosis

2012 ◽  
Vol 9 (1) ◽  
pp. 19-23 ◽  
Author(s):  
S B Marahatta ◽  
S Gautam ◽  
S Dhital ◽  
N Pote ◽  
A K Jha ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Deoxyribonucleic Acid ◽  
Fragment Length Polymorphism ◽  
Drug Susceptibility ◽  
Specific Gene ◽  
Katg Gene ◽  
Tb Treatment ◽  
High Prevalence ◽  
Pcr Rflp ◽  
Polymerase Chain

Background Isoniazid (INH) together with Rifampicin (RFP) forms the cornerstone of a short chemotherapy course for tuberculosis (TB) treatment. Mutation at codon 315 of katG gene is most prevalent in isoniazid resistant Mycobacterium tuberculosis (MTB) and is high in area with high TB incidence. Polymerase Chain Reaction Restriction Fragment Length Polymorphism (PCR-RFLP) has been found to be a reliable and effective tool for the identification of the specific gene alteration. Objective The objective of this study was to screen Ser315Thr mutation of KatG gene of INH resistant MTB strain by PCR-RFLP technique. Methods Altogether 37 INHr MTB isolates obtained from German Nepal Tuberculosis Project (GENETUP) Kathmandu Nepal was included in the study. Deoxyribonucleic Acid (DNA) extraction was performed according to protocol of SORPOCLEAN™ from the culture isolates. Amplification of the fragment with katG codon 315 was performed in a Biometra Thermocycler using primers. The amplified fragment was cleaved with MspI. The restriction fragments obtained were electrophoresed in a 2% agarose gel and were visualized using transilluminator. Results The katG Ser315Thr mutation was observed in 23 (62.2%) out of 37 INH resistant isolates. The drug susceptibility profile of INHr MTB isolates showed all isolates to be resistant to INH and RFP whereas 26 and 27 MTB isolates were resistant to Ethambutol (EMB) and Streptomycin (S) respectively. Seventeen (17) patients were harbouring katG gene mutated strain among Ethambutol and Streptomycin resistant cases. Conclusion The study identified high prevalence of Ser315Thr mutation in katG. The isolates harbouring this mutation were also simultaneously resistant to RFP. Ser315Th could be a potential genetic marker for predicting MDR-TBhttp://dx.doi.org/10.3126/kumj.v9i1.6256 Kathmandu Univ Med J 2011;9(1):19-23

Elaphostrongylus cervi in a population of red deer (Cervus elaphus) and evidence of cerebrospinal nematodiasis in small ruminants in the province of Varese, Italy

2010 ◽  
Vol 85 (3) ◽  
pp. 313-318 ◽  
Author(s):  
E.G. Alberti ◽  
G. Gioia ◽  
G. Sironi ◽  
S. Zanzani ◽  
P. Riccaboni ◽  
...  
Keyword(s):  
Cervus Elaphus ◽  
Red Deer ◽  
Small Ruminants ◽  
Domestic Ruminants ◽  
Faecal Samples ◽  
High Prevalence ◽  
Internal Parasites ◽  
History Of ◽  
Central Nervous Systems ◽  
Polymerase Chain

AbstractThirty-one faecal samples were collected from red deer in the northern area of Varese, in the Italian region of Lombardy, between August and October 2008. The animals had either been hunted or accidently killed. Examination for internal parasites showed a prevalence of 45.2% for Elaphostrongylus cervi larvae and species identification was confirmed by polymerase chain reaction (PCR). Ninety-seven faecal samples were also collected from two goat flocks grazing in the same area between December 2007 and May 2008. These showed a prevalence of 74.7% for lungworms. Furthermore, the central nervous systems from five goats and one sheep from this area with a history of neurologically related lameness were examined. Histopathology confirmed E. cervi cerebro- spinal nematodiasis in five cases out of six. This study demonstrates E. cervi transmission from wild to domestic ruminants when the animals graze in the same area, and the possible occurrence of clinical disease in infected goats and sheep associated with high prevalence in deer.

Molecular identification of hookworm isolates from stray dogs, humans and selected wildlife from South Africa

2019 ◽  
Vol 94 ◽  
Author(s):  
P. I. Ngcamphalala ◽  
J. Lamb ◽  
S. Mukaratirwa
Keyword(s):  
South Africa ◽  
Polymerase Chain Reaction ◽  
Fragment Length Polymorphism ◽  
Domestic Animals ◽  
Chain Reaction ◽  
Stray Dogs ◽  
5.8S Rrna ◽  
Faecal Samples ◽  
Pcr Rflp ◽  
Polymerase Chain

Abstract There is a paucity of information on hookworm species in humans, domestic animals and wildlife in southern Africa. Our study aimed to identify hookworm species from stray dogs, humans, and selected wildlife from South Africa. A total of 356 faecal samples were screened for the presence of hookworm-like eggs and subsequently coproculture from the positive samples was carried out to obtain larvae. Hookworm-like eggs were detected in 23.03% (82/356) of samples. Of these samples, 78/296 were from dogs, 3/50 from humans and 1/10 from wildlife. DNA was then isolated from the larvae of 55 positive samples, which were subjected to polymerase chain reaction (PCR), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing of the nuclear ribosomal internal transcribed spacer (ITS1) and 5.8S rRNA region. Presence of Ancylostoma caninum, A. braziliense and A. ceylanicum-like species was recorded in stray dogs and A. caninum was recorded in wildlife and humans, using PCR-RFLP. Although PCR-RFLP results pointed to the presence of A. ceylanicum, we did not get a sequence that matched with A. ceylanicum from GenBank. This may have been due to the low proportion of A. ceylanicum larvae in our samples. Twenty-two of the 27 positive amplicons from stray dogs matched with A. caninum, three with A. braziliense and two had mixed infections of A. braziliense and A. caninum. Sequences from a lion and three humans matched with A. caninum. This is the first confirmation of a patent A. caninum infection in humans as evidenced by the presence of eggs in faeces, with the subsequent larvae from coproculture being identified as A. caninum.

Molecular differentiation of three canine and feline hookworms in South China through HRM analysis

2018 ◽  
Vol 93 (2) ◽  
pp. 159-165 ◽  
Author(s):  
M.W. Wang ◽  
X.X. Yan ◽  
J.X. Hang ◽  
X.L. Shi ◽  
Y.Q. Fu ◽  
...  
Keyword(s):  
South China ◽  
High Resolution Melting ◽  
Fragment Length Polymorphism ◽  
Melting Curve ◽  
Epidemiological Survey ◽  
Clinical Samples ◽  
Chain Reaction ◽  
Zoonotic Risk ◽  
Pcr Rflp ◽  
Polymerase Chain

AbstractTo investigate the prevalence of canine and feline hookworms in South China, and to assess the risk of zoonotic hookworms to humans, one pair of primers (HRM-F/HRM-R) was designed to establish a high-resolution melting (HRM) method based on internal transcribed spacer 1 (ITS-1) rDNA for the detection of Ancylostoma ceylanicum, A. caninum and A. tubaeforme infection. The results showed that the HRM for the three hookworms produced different melting-curve profiles, where melting temperature (Tm) values were 84.50°C for A. ceylanicum, 82.25°C for A. caninum and 81.73°C for A. tubaeforme, respectively. The reproducibility of intra- and inter-assay melting curves was almost perfect. The lowest concentration detected was about 5.69 ×10−4 g/μl. The HRM detection results from 18 canine and feline hookworm samples were in complete accordance with their sequencing results. The HRM method was more sensitive than the polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) technique in the detection of 98 clinical samples. It is concluded that the HRM method can differentiate between A. ceylanicum, A. caninum, A. tubaeforme and their mixed infections, which may provide important technical support for the zoonotic risk assessment and molecular epidemiological survey of canine and feline hookworms.

scholarly journals RAPID PCR–BASED DETECTION OPTIMIZATION OF PORCINE DNA IN GELATIN CAPSULE SHELL

2018 ◽  
Vol 10 (6) ◽  
pp. 217
Author(s):  
Amalia Sitti Khayyira ◽  
Viktoria Mardhika Estepane ◽  
Amarila Malik
Keyword(s):  
Molecular Detection ◽  
Universal Primers ◽  
Gelatin Capsule ◽  
Duplex Pcr ◽  
Rapid Pcr ◽  
Capsule Shell ◽  
Pcr Rflp ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Bovine Species

Objective: This study was conducted to optimize the genomic deoxyribonucleic acid (DNA) based molecular detection of gelatin derived from porcine by performing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and duplex PCR method employing cyt B gene.Methods: Optimization was carried out for DNA extraction, PCR conditions, and the sensitivity of the PCR-RFLP method. Due to the very low DNA trace in gelatin after the various manufacturing process, the extraction was optimized to obtain sufficient DNA which was visible on the agarose gel. PCR-RFLP was carried out using universal primers and BsaJI restriction enzyme, and duplex PCR was carried out using two sets of porcine-specific primers. Porcine and bovine DNA were mixed in various concentration to confirm sensitivity of both methods, i.e. 100%, 50%, 10%, 1%, 0.5%, 0.1%, 0.05%, and 0.01%Results: Both methods, PCR-RFLP, and Duplex PCR, were able to detect as low as 0.01% porcine DNA, indicated by the presence of porcine DNA amplicon bands (131 bp and 228 bp for PCR-RFLP, 212 bp and 398 bp for duplex PCR). Although DNA bands presented in low intensity, identification of porcine and bovine species and estimation of DNA quantities were possible.Conclusion: Both conventional PCR methods, i.e. PCR-RFLP and Duplex PCR, were sensitive, specific, and suitable as a rapid initial detection method for molecular detection of porcine in gelatin capsule shells.

scholarly journals Molecular Screening of Salmonella sp. from fecal sample of Sparrows (Passer domesticus) in Yogyakarta, Indonesia

2021 ◽  
Vol 33 ◽  
pp. 07003
Author(s):  
Marla Anggita ◽  
Okti Herawati ◽  
Sidna Artanto
Keyword(s):  
Target Genes ◽  
Passer Domesticus ◽  
Local Area ◽  
Zoonotic Diseases ◽  
Molecular Screening ◽  
Chain Reaction ◽  
Intervention Measures ◽  
Faecal Samples ◽  
Pcr Method ◽  
Polymerase Chain

Wild birds is one of the reservoir agent of some of various zoonotic diseases. The study was aim to see the potential of sparrow as the reservoir agent of Salmonella sp. using polymerase chain reaction (PCR) method. We detected the invA gene of Salmonella sp. from faecal sample of sparrows (Passer domesticus) in local area of Yogyakarta, Indonesia. A total of 30 faecal dropping samples were collected from sparrows. DNA was extracted from the faecal samples, then amplified by PCR for the target genes. The amplicons were electrophorized to see the visualization of DNA on the agarose gel. The result showed the prevalence of the positive result of Salmonella sp. was 3,3%. The study indicated that sparrows can spread zoonotic pathogens and this necessitates monitoring for the epidemiologic status of these pathogens among birds, also applying the appropriate intervention measures to prevent the transmission of zoonotic diseasesfrom birds to humans.

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