scholarly journals Serological diagnosis and molecular characterization of Leptospira spp. in the blood and urine of bovine females from refrigerated slaughterhouses

2018 ◽  
Vol 39 (3) ◽  
pp. 1125
Author(s):  
Aliny Fernanda de Oliveira ◽  
Roberta Torres Chiderolli ◽  
Luciano Seraphim Gasques ◽  
Arianne Peruzo Pires Gonçalves ◽  
Érica Dourado Neves ◽  
...  
Keyword(s):  
One Health ◽  
Preventive Measures ◽  
Agglutination Test ◽  
Economic Losses ◽  
Chain Reaction ◽  
Serum Samples ◽  
Leptospira Spp ◽  
Blood And Urine Samples ◽  
Polymerase Chain

Leptospirosis is an important socioeconomic disease in humans, as well as in domestic and wild animals, being caused by Leptospira spp. Bovine animals are considered reservoirs of this disease, because they intermittently disseminate the bacteria into the environment through their urine. In this way, the cattle an important source of Leptospira infection. The objective of this study was to detect Leptospira spp. antibodies and DNA in bovine females from two refrigerated slaughterhouses in the microregion of Umuarama, Paraná, Brazil. In particular, blood and urine samples from 52 crossbred bovine females older than 36 months from the two slaughterhouses were used. The microscopic agglutination test (MAT) was used to detect leptospiral antibodies, and the polymerase chain reaction (PCR) and subsequent sequencing were used to detect Leptospira DNA. The MAT yielded 22 (42.3%) serum samples considered reagent, while the nested PCR test resulted in one amplified sample (1.9%) of 289 bp. This single sample was then amplified again using primers for the SecY gene (549 bp). Sequencing of this gene characterized the bacteria as L. borgpetersenii that were similar to the serovar Hardjo of the genotype Hardjobovis. This is the first molecular confirmation of Hardjobovis-like L. borgpetersenii in the urine of crossbred bovine females older than 36 months from slaughterhouses in the microregion of Umuarama. This study’s results show that it is important to combine serological and molecular diagnosis in the detection of Leptospira spp. Therefore, both methods were used to improve our understanding of the epidemiology of this disease in bovine animals from the microregion of Umuarama. In addition, the analysis informed the subsequent adoption of preventive measures and educational One Health actions to prevent economic losses related to the herd, as well as social losses related to workers and the environment.

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 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 First Isolation and Molecular Typing of Pathogenic and Intermediate Leptospira Species from Urine of Symptomatic Dogs

2021 ◽  
Vol 8 (12) ◽  
pp. 304
Author(s):  
Ivana Piredda ◽  
Loris Bertoldi ◽  
Giuseppe Benvenuto ◽  
Bruna Palmas ◽  
Aureliana Pedditzi ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Zoonotic Risk ◽  
Leptospira Spp ◽  
Blood And Urine Samples ◽  
Healthy Dogs ◽  
Polymerase Chain ◽  
Sequence Types ◽  
Leptospira Species

Aim of this study was to evaluate, the presence and diversity of Leptospira spp. in blood and urine samples collected from 175 owned-dogs from Sardinia, Italy. After determination of leptospiral infection by microscopic agglutination test (MAT), urine from MAT-positive dogs were examined by real-time polymerase chain reaction (lipL32 rt-PCR) and then isolated by culture. In order to characterize obtained serovars, positive cultures were then subjected to 16S rRNA and secY sequencing, phylogenetic analysis and Multilocus Sequence Typing (MLST). Results showed that seven dogs (4%; 95% CI: 0–55) had Leptospira DNAs in their urine and five strains were isolated from urine cultures. The three different sequence types (ST17, ST198 and ST24) belonging to Leptospira interrogans genomospecies identified by MLST analyses in this study, confirmed that the leptospiral infection was widespread in Sardinian dogs. We also reported the first characterization of a new Leptospira spp. isolated from urine of one dog living in the study area. Whole genome sequencing and phylogenetic analysis, confirmed that this genospecies was closely related to Leptospira hovindhougenii, an intermediate Leptospira spp. with unknown pathogenicity previously isolated from a rat in Denmark. Further studies are required to clarify whether healthy dogs that shed leptospires in their urine could represent a zoonotic risk for humans in this region.

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 SEROLOGICAL AND MOLECULAR SURVEY OF Leptospira spp. AMONG CART HORSES FROM AN ENDEMIC AREA OF HUMAN LEPTOSPIROSIS IN CURITIBA, SOUTHERN BRAZIL

2014 ◽  
Vol 56 (6) ◽  
pp. 473-476 ◽  
Author(s):  
Mariane Angélica Finger ◽  
Ivan Roque de Barros Filho ◽  
Christian Leutenegger ◽  
Marko Estrada ◽  
Leila Sabrina Ullmann ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Endemic Area ◽  
Agglutination Test ◽  
Southern Brazil ◽  
Chain Reaction ◽  
Human Leptospirosis ◽  
Leptospira Spp ◽  
Polymerase Chain ◽  
Positive Correlations

Introduction: Cart horses are a re-emerging population employed to carry recyclable material in cities. Methods: Sixty-two horses were sampled in an endemic area of human leptospirosis. The microscopic agglutination test (MAT) and real-time polymerase chain reaction (qPCR) were performed. Results: A seropositivity of 75.8% with serovar Icterohaemorrhagiae in 80.8% of the horses was observed. Blood and urine were qPCR negative. MAT showed positive correlations with rainfall (p = 0.02) and flooding (p = 0.03). Conclusions: Although horses may be constantly exposed to Leptospira spp. in the environment mostly because of rainfall and flooding, no leptospiremia or leptospiruria were observed in this study.

scholarly journals Occurrence of anti-Toxoplasma gondii antibodies and parasite DNA in backyard chicken breeding in Northeast, Brazil

2016 ◽  
Vol 25 (1) ◽  
pp. 105-108 ◽  
Author(s):  
Marcela Fernanda Torres Samico Fernandes ◽  
Erika Fernanda Torres Samico Fernandes Cavalcanti ◽  
José Givanildo da Silva ◽  
André da Rocha Mota ◽  
Orestes Luiz de Souza Neto ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Preventive Measures ◽  
Immunofluorescence Assay ◽  
Northeast Brazil ◽  
Chain Reaction ◽  
Serum Samples ◽  
Raw Meat ◽  
Polymerase Chain ◽  
Backyard Chickens ◽  
Backyard Chicken

Abstract The aim of the present study was to investigate the occurrence of anti-Toxoplasma gondii antibodies and parasite DNA in backyard chickens bred in the metropolitan area of Recife, Brazil. In total, 212 serum samples were collected from 16 properties, and 12 backyard chickens were collected in the six sanitary districts of Recife. An indirect immunofluorescence assay (IFA) was used to investigate the occurrence of anti-Toxoplasma gondii antibodies. Polymerase chain reaction (PCR) was used to detect T. gondii DNA in brain, heart, liver and lung specimens. Of the samples analyzed by serology, 86/212 (40.56%) were positive; of the samples analyzed by PCR, 2/12 (16.7%) were positive, with both samples positive by both tests (serological and molecular). The presence of antibody anti-T. gondii and parasite DNA in tissues of these animals are worrying aspects for public health because there is a risk of transmission of the parasite to humans through eating undercooked or raw meat. Based on the results, the adoption of preventive measures to prevent the cats access to the chickens creations should be encouraged, since these animals were identified in most of the studied properties.

scholarly journals Infections of swine caused by Leptospira serovars of serogroup Sejroe - possibilities of recognition with the use of PCR

2014 ◽  
Vol 58 (4) ◽  
pp. 521-526 ◽  
Author(s):  
Bernard Wasiński
Keyword(s):  
Dna Sequences ◽  
Agglutination Test ◽  
Urine Samples ◽  
Chain Reaction ◽  
Serum Samples ◽  
Two Samples ◽  
Leptospira Serovars ◽  
Polymerase Chain ◽  
Primer Sets ◽  
Leptospira Species

Abstract The aim of this study was to confirm the presence of leptospires from the Sejroe serogroup in aborting seropositive sows and to identify which serovars of this serogroup are responsible for the infection. Serum and urine samples from 20 aborting sows, reared in five farms infected with the Sejroe serogroup, were submitted for examination. Serum samples were examined by the microscopic agglutination test (MAT) and urine samples by polymerase chain reaction (PCR) with primer sets specific for genus Leptospira, species L. borgpetersenii, the Sejroe serogroup, and the Hardjo-bovis genotype. All the examined serum samples showed titres specific to the Sejroe serogroup, ranging from 100 to 6400. PCR indicated the presence of Leptospira genus-specific DNA sequences in all urine samples examined. DNA sequences specific for both L. borgpetersenii and the Sejroe serogroup were found in 12 of these samples. Additionally, the sequences specific only for L. borgpetersenii were found in five urine samples and sequences for the Sejroe serogroup in two samples. The DNA sequence specific for the Hardjo-bovis genotype was not found in the urine samples. PCR confirmed the results of the MAT, displayed the presence of Leptospira sp. DNA in the urine of all the sows examined, proved the affinity of the detected leptospires to the Sejroe serogroup or at least to L. borgpetersenii, and excluded the Hardjo serovar as the reason for the infections described in the sows.

scholarly journals Molecular Identification of Leptospira interrogans in Naturally Infected Cows from a Rural Property in the Border Region - Case Report

2020 ◽  
Vol 41 (4) ◽  
pp. 1433
Author(s):  
Giovani Batista Pastre ◽  
Isabela Carvalho dos Santos ◽  
Robson Michael Delai ◽  
Edinalva Madalena de Almeida Mota ◽  
Lidiane Nunes Barbosa ◽  
...  
Keyword(s):  
Production Efficiency ◽  
Dairy Farm ◽  
Border Region ◽  
Antibody Detection ◽  
Chain Reaction ◽  
Border Regions ◽  
Leptospira Spp ◽  
Blood And Urine Samples ◽  
Polymerase Chain ◽  
Serum Agglutination

The aim of this study was to investigate the antibodies and DNA of Leptospira spp. isolated from infected cattle in a small rural dairy farm in a border region between Brazil and Paraguay. Blood and urine samples were collected from 50 Holstein cows aged between 1 and 15 years. The diagnostic tests performed were microscopic serum agglutination for antibody detection and polymerase chain reaction for Leptospira spp. detection. Out of the samples analyzed, 48% were MAT positive with titers ranging from 100 to 400, and the most prevalent antibody was to the serovar Hardjo. One serum sample was amplified to 549 bp for the sec y gene, and sequencing identified it as L. interrogans. This is the first report from northwestern Paraná (PR) State of L. interrogans identification in naturally infected milk cattle. Thus, based on these results, to enhance production efficiency, new serological and molecular studies on dairy cattle from border regions are required to characterize the epidemiology of possible genotypes and their consequences in affected herds.

scholarly journals Partial characterization of Prunus Necrotic Ringspot Virus on apple in EGYPT 

2021 ◽  
Author(s):  
Aly M. Abdel-Salam ◽  
Samah A. Mokbel
Keyword(s):  
Binding Assay ◽  
Ringspot Virus ◽  
Economic Losses ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Prunus Necrotic Ringspot Virus ◽  
Polymerase Chain ◽  
Severe Isolate ◽  
Microscopy Examination

Abstract A severe isolate of Prunus necrotic ringspot virus (PNRSV) was isolated from apple orchards in the vicinity of Nubaria city, Beheira governorate, Egypt. Infected-apple trees showed chlorotic, necrotic ringspots, and shoot holes on leaves. Severely infected- trees withered, became useless, and were removed causing severe economic losses. Reverse transcriptase (RT) polymerase chain reaction (PCR), RT-PCR, using degenerate primer pair for the coat protein (CP) gene of Ilarvirus amplified products similar to those produced from peach and apricot isolates of PNRSV-infecting stone fruits). Dot blotting immuno-binding assay (DBIA showed a positive reaction between PNRSV-infected apple sap and an Egyptian antiserum for PNRSV. Purified preparation from infected leaves, using the electro-elution technique yielded nucleoprotein, which had Amax and Amin at 260 and 240 nm respectively. Electron microscopy examination showed spherical virions with ca. 26 nm in diameter.

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