Molecular Detection of Leptospira Species Serotypes in Iranian Stray Dogs

2019 ◽  
Author(s):  
Saam Torkan ◽  
Hassan Momtaz
Keyword(s):  
Public Health ◽  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Serum Samples ◽  
Blood Samples ◽  
Stray Dogs ◽  
Wide Range ◽  
Leptospira Spp ◽  
Polymerase Chain ◽  
Leptospira Species

Background and Aims: Leptospirosis is a spirochetal disease with public health importance globally. This disease affects a wide range of domestic and wild animals. Dogs are one of the species most sensitive to Leptospira canicola and Leptospira icterohaemorrhagiae. The present study was concluded to evaluate the prevalence rate of Leptospira species and L. canicola and L. icterohaemorrhagiae serovars in Iranian stray dogs. Materials and Methods: One-hundred and twenty blood samples were first taken from stray dogs. Then the samples were transferred to the laboratory. Sera were extracted from blood samples and genomic DNA was extracted. DNA samples were subjected to conventional polymerase chain reaction. Positive samples for Leptospira spp. were analyzed for presence of L. canicola and L. icterohaemorrhagiaeserovars using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results: Nine samples out of 120 serum samples (7.5%) were positive for the flagella gene of the Leptospira spp. Prevalence of Leptospira spp. in serum samples of male and female dogs were 5.4% and 10.86%, respectively. Prevalence of L. canicola and L. icterohaemorrhagiae serovars were 55.55% and 33.33%, respectively. We found that 11.11% of samples were positive for both serovars. Two to three and 3-4 year old dogs had the highest prevalence of Leptospira spp. Conclusions: The considerable prevalence of leptospirta spp. and also their zoonotic serovars among Iranian stray dogs represented an important public health issue regarding the contact of healthy human with these dogs. Identification of infected dogs and their vaccination can inhibit the distribution of Leptospira spp.

scholarly journals Leptospira Detection in Cats in Spain by Serology and Molecular Techniques

2020 ◽  
Vol 17 (5) ◽  
pp. 1600 ◽  
Author(s):  
Andrea Murillo ◽  
Rafaela Cuenca ◽  
Emmanuel Serrano ◽  
Goris Marga ◽  
Ahmed Ahmed ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Human Infection ◽  
Molecular Techniques ◽  
Antibody Prevalence ◽  
Chain Reaction ◽  
Pathogenic Leptospira ◽  
Blood Samples ◽  
Polymerase Chain ◽  
Positive Results ◽  
Leptospira Species

Leptospirosis is the most neglected widespread zoonosis worldwide. In Spain, leptospirosis reports in people and animals have increased lately. Cats can become infected with Leptospira, as well as be chronic carriers. The aim of this study was to determine serological antibody prevalence against Leptospira sp., blood DNA, and shedding of DNA from pathogenic Leptospira species in the urine of cats in Spain. Microagglutination tests (MAT) and blood and urine TaqMan real-time polymerase chain reaction (PCR) were performed. Leptospira antibodies were detected in 10/244 cats; with 4.1% positive results (95% confidence interval (CI): 2.1–7.18%). Titers ranged from 1:20 to 1:320 (serovars Ballum; Bataviae; Bratislava; Cynopteri; Grippotyphosa Mandemakers; Grippotyphosa Moskva; Pomona; and Proechimys). The most common serovar was Cynopteri. Blood samples from 1/89 cats amplified for Leptospira DNA (1.12%; 95% CI: 0.05–5.41%). Urine samples from 4/232 cats amplified for Leptospira DNA (1.72%; 95% CI: 0.55–4.10%). In conclusion free-roaming cats in Spain can shed pathogenic Leptospira DNA in their urine and may be a source of human infection. Serovars not previously described in cats in Spain were detected; suggesting the presence of at least 4 different species of pathogenic leptospires in the country (L. borgpetersenii; L. interrogans; L. kirschneri; and L. noguchii).

scholarly journals Detection of Brucella sp. and Leptospira sp. in dogs using conventional polymerase chain reaction

2014 ◽  
Vol 58 (4) ◽  
pp. 527-531
Author(s):  
Faham Khamesipour ◽  
Abbas Doosti ◽  
Mohsen Fard Emadi ◽  
Babafela Awosile
Keyword(s):  
Polymerase Chain Reaction ◽  
Age Groups ◽  
Conventional Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Blood Samples ◽  
Stray Dogs ◽  
Supplementary Method ◽  
Significant Difference ◽  
Polymerase Chain ◽  
First Time

Abstract The study was conducted to detect Brucella sp. and Leptospira sp. in blood samples of dogs in Isfahan and Shahrekord province in Iran. A total of 94 blood samples were collected from dogs of different breed, age, sex, and dogs’ type (stray or nonstray). The samples were examined using conventional polymerase chain reaction (PCR). Fourteen (14.89%) dogs were positive for Brucella sp. and 18 (19.15%). dogs for Leptospira sp. There were no significant differences between the prevalence of the pathogens, provinces, sex, and age groups (P > 0.05). However, there was a statistically significant difference in prevalence of Brucella sp. and Leptospira sp. between stray and non-stray dogs (P < 0.0001; χ2 = 30.3767). The study also demonstrated that PCR was successfully used for the first time in Iran for the detection of Brucella sp. and Leptospira sp. in blood samples of dogs. Therefore, we recommend the PCR as a supplementary method with other commonly recognised methods (e.g. serological methods) for the diagnosis of subclinical infections with the microorganisms. Strict measures for the control of stray dogs are also highly recommended.

scholarly journals Blanding’s turtles (Emydoidea blandingii) as a reservoir for Leptospira spp

2019 ◽  
Author(s):  
Kelly E. Rockwell ◽  
Dan Thompson ◽  
Carol Maddox ◽  
Mark A. Mitchell
Keyword(s):  
Polymerase Chain Reaction ◽  
Aquatic Ecosystem ◽  
Urine Samples ◽  
Leptospira Interrogans ◽  
Chain Reaction ◽  
Natural Conditions ◽  
Blood Samples ◽  
Emydoidea Blandingii ◽  
Leptospira Spp ◽  
Polymerase Chain

AbstractLeptospira spp. are re-emerging zoonotic pathogens. Previous research has found that Blanding’s turtles (Emydoidea blandingii) experimentally infected with Leptospira interrogans shed leptospires in their urine, suggesting that they could play a role in transmitting pathogen within an aquatic ecosystem. This study investigated whether a population of wild Blanding’s turtles known to be exposed to Leptospira spp. actively shed the pathogen under natural conditions. Blood samples were collected for serologic testing and to assess the health of the turtles. Free catch urine was collected for polymerase chain reaction (PCR) testing. All turtles were seropositive for Leptospira spp. and 73.5% (25/34) of the urine samples were PCR positive. All animals appeared clinically healthy and showed no apparent signs of disease. This study confirms that wild Blanding’s turtles can actively shed Leptospira spp. in their urine and suggests that they may play a role in the epidemiology of this disease in habitats in which they reside.

scholarly journals Frequency of Leptospira spp. in sheep from Brazilian slaughterhouses and its association with epidemiological variables

2012 ◽  
Vol 32 (3) ◽  
pp. 194-198 ◽  
Author(s):  
Rodrigo Costa da Silva ◽  
Veruska Maia da Costa ◽  
Fabio Hiroto Shimabukuro ◽  
Virgínia Bodelão Richini-Pereira ◽  
Benedito Donizete Menozzi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Bacterial Growth ◽  
Agglutination Test ◽  
The State ◽  
Rodent Control ◽  
Chain Reaction ◽  
Serum Samples ◽  
Leptospira Spp ◽  
Positive Serum ◽  
Polymerase Chain

Leptospirosis is a worldwide anthropozoonosis that infects livestock, including sheep as the carriers to other animals and humans. The present study aimed to determine the prevalence of Leptospira spp. in sheep from two slaughterhouses in the state of São Paulo, Brazil and its association with epidemiological variables. Serum samples from 182 sheep were evaluated for Leptospira spp. antibodies by microscopic agglutination test (MAT). Results indicated 34/182 (18.68%; CI95% 13.70-24.98%) positive serum samples, mainly to the serovar Copenhageni (17/34; 50%; CI95% 33.99-66.01%). Bacterial growth in the Fletcher medium was detected for 13/34 (38.24%; CI95% 23.87-55.08%) animals, and confirmed by Polymerase Chain Reaction (PCR) and sequencing for only two kidney samples from two animals. Thus, treatment and vaccination of sheep, besides rodent control, can be useful to prevent the infection in the studied region since sheep are important Leptospira spp. carriers, and its transmission to slaughterhouse workers is mainly through the manipulation of visceral tissues.

scholarly journals Determine the prevalence of Brucella spp. and Leptospira spp. in blood samples by multiplex polymerase chain reaction collected from cattle, sheep and goats in herds located in provinces of Iran

2014 ◽  
Vol 4 (1) ◽  
Author(s):  
Faham Khamesipour ◽  
Shahin Nejat Dehkordi ◽  
Taghi Taktaz Hafshejani ◽  
Elahe Tajbakhsh ◽  
Shahrzad Azizi
Keyword(s):  
Polymerase Chain Reaction ◽  
Multiplex Polymerase Chain Reaction ◽  
Animal Health ◽  
Chain Reaction ◽  
Routine Diagnostics ◽  
Blood Samples ◽  
Sheep And Goats ◽  
Polymerase Chain ◽  
Breeding Farms ◽  
Leptospira Species

Leptospirosis and brucellosis are common zoonosis that affect many species of mammals mostly causing economical losses. Further, very important fact is huge danger for human and animal health around the world. The purpose of the study is to determine the prevalence of <em>Brucella</em> spp. and <em>Leptospira</em> spp. using multiplex polymerase chain reaction (mPCR) method, in blood samples collected from cattle, sheep and goats. In this study, a total number of 250 blood samples (5 cc of blood with ethilen diamin tetra asetic acid) were collected randomly from 100 cattle, 80 sheep and 70 goats located on 6 herds in Chaharmahal Va Bakhtiari and Esfahan provinces, Iran. After DNA extraction and setting of mPCR for <em>Brucella</em> spp. and <em>Leptospira</em> spp. mPCR products were screened. The DNA of these microorganisms was detected by multiplex PCR from 31 and 21 out of 100 cattle, respectively. Four of 70 goat’s blood samples from goat breeding farms were positive for <em>Leptospira</em> spp. and 11 were positive for <em>Brucella</em> spp. Out of 80 sheep blood samples 23 were positive for <em>Brucella</em> spp. and 14 for <em>Leptospira</em> spp. The results of the present study show ruminant as an important reservoir for transmission of these zoonotic diseases to humans in Iran. mPCR has the ability to concurrently detect both Brucella and Leptospira species from blood samples of ruminants. The convenience and the possibility of detection of both bacteria at a time, strongly support the use of this mPCR for routine diagnostics.

scholarly journals Evidence of exposure to Leptospira spp. in dogs at a zoonosis control center in Brazil

2020 ◽  
Vol 18 ◽  
pp. 1
Author(s):  
Mírian Da Rocha Albuquerque ◽  
Thalita Amaral dos Reis ◽  
Katarine De Souza Rocha ◽  
Jacqueline Da Silva Brito ◽  
Gleiciane Schupp De Sena Mesquita ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Whole Blood ◽  
Agglutination Test ◽  
Microscopic Agglutination Test ◽  
Chain Reaction ◽  
Serum Samples ◽  
Control Center ◽  
Leptospira Spp ◽  
Polymerase Chain ◽  
Prior Contact

Evidence of exposure to Leptospira spp. in dogs housed in the kennel of the Zoonosis Control Center of Belém, Pará, Brazil, was investigated. Whole blood and serum samples from 145 dogs were investigated using the polymerase chain reaction (PCR) and microscopic agglutination test (MAT), respectively. A total of 64.14% of the dogs were found to be seropositive for Leptospira spp., with the most frequent serogroup being Djasiman (39.73%). However, PCR results revealed that all of the dogs were negative for Leptospira spp. DNA. Although the results of the study suggest the animals did not currently have leptospires in blood, they only show circulating anti-Leptospira spp. antibodies, implying prior contact with the bacteria.

scholarly journals Development of a Genoserotyping Method for Salmonella Infantis Detection on the Basis of Pangenome Analysis

2020 ◽  
Vol 9 (1) ◽  
pp. 67
Author(s):  
Seung-Min Yang ◽  
Jiwon Baek ◽  
Eiseul Kim ◽  
Hyeon-Be Kim ◽  
Seyoung Ko ◽  
...  
Keyword(s):  
Public Health ◽  
Polymerase Chain Reaction ◽  
Bacterial Species ◽  
Cost Effective ◽  
The Other ◽  
Gene Marker ◽  
Diagnostic Assay ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Substantial Economic Burden

In recent years, Salmonella Infantis has become a predominant serovariant in clinical and poultry isolates, thereby imposing a substantial economic burden on both public health and the livestock industry. With the aim of coping with the steep increase in serovar Infantis prevalence, a polymerase chain reaction (PCR)-based rapid and accurate diagnostic assay was developed in this study through pangenome profiling of 60 Salmonella serovars. A gene marker, SIN_02055, was identified, which is present in the S. Infantis genome but not in the pangenome of the other serovars. Primers specific to SIN_02055 were used to accurately detect serovar Infantis, and to successfully differentiate Infantis from the other 59 serovars in real-time PCR with a R2 of 0.999 and an efficiency of 95.76%. The developed method was applied to 54 Salmonella strains belonging to eight dominant serovars, and distinguished Infantis from the other seven serovars with an accuracy of 100%. The diagnostic primer set also did not show false positive amplification with 32 strains from eight non-Salmonella bacterial species. This cost-effective and rapid method can be considered an alternative to the classic serotyping using antisera.

Comparison of Droplet Digital Polymerase Chain Reaction (ddPCR) and Real-Time Quantitative Polymerase Chain Reaction (qPCR) in Detecting Neonatal Invasive Fungal Infections

2021 ◽  
Vol 11 (3) ◽  
pp. 373-379
Author(s):  
Huitao Li ◽  
Xueyu Chen ◽  
Xiaomei Qiu ◽  
Weimin Huang ◽  
Chuanzhong Yang
Keyword(s):  
Polymerase Chain Reaction ◽  
Limit Of Detection ◽  
Quantitative Polymerase Chain Reaction ◽  
Diagnostic Methods ◽  
Chain Reaction ◽  
Fungal Isolation ◽  
Blood Samples ◽  
Fungal Strains ◽  
Polymerase Chain ◽  
Digital Polymerase Chain Reaction

Invasive fungal infection (IFI) is the leading cause of death in neonatal patients, yet the diagnosis of IFI remains a major challenge. At present, most IFI laboratory diagnostic methods are based on classical, but limited, methods such as fungal isolation and culture and histopathological examination. Recently, quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) technology have been adopted to quantify nucleic-acid identification. In this study, we established qPCR and ddPCR assays for IFI diagnosis and quantification. qPCR and ddPCR were carried out using identical primers and probe for the amplification of 18S rRNA. Assay results for three fungal strains were positive, whereas ten non-fungal strains had negative results, indicating 100% specificity for both ddPCR and qPCR methods. Genomic DNA of Candida albicans was tested after a serial dilution to compare the sensitivity of the two PCR methods. The limit of detection of ddPCR was 3.2 copies/L, which was a ten-fold increase compared with that of the qPCR method (32 copies/L). Blood samples from 127 patients with high-risk factors and clinical symptoms for IFI were collected from a NICU in Shenzhen, China, and analyzed using qPCR and ddPCR. Thirty-four blood samples from neonates had a proven or probable diagnosis of IFI, and 25 of these were positive by qPCR, whereas 30 were positive by ddPCR. Among the 93 blood samples from neonates who had a possible IFI or no IFI, 24 were positive using qPCR, and 7 were positive using ddPCR. In conclusion, ddPCR is a rapid and accurate pan-fungal detection method and provides a promising prospect for IFI clinical screening.

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