ABSTRACT
Canine Leproid Granuloma Syndrome (CLGS), also known as canine leprosy, is a cutaneous nodular infectious disease caused by Mycobacterium sp.. Despite being reported worldwide, it is still quite unknown and underdiagnosed. Diagnosis may be achieved by cytopathology or histopathology of skin lesions, but identification of the infectious agent is complex, since bacterial in vitro growth is not possible, relying upon molecular techniques such as PCR to confirm Mycobacterium DNA in the sample. We report a CLGS case in Niteroi, Rio de Janeiro state, Brazil, diagnosed by cytopathology and submitted to molecular identification of the agent. PCR amplification of hsp65 gene was performed and revealed 100% genetic homology to M. murphy strain. This is the first CLGS report with molecular identification in Rio de Janeiro state, and this finding should raise awareness about CLGS as a differential diagnosis among granulomatous skin diseases in this region.(AU)
A síndrome de granuloma leproide canino (SGLC), também conhecida como lepra canina, é uma doença infecciosa cutânea nodular causada por Mycobacterium sp. Apesar de ser relatada mundialmente, ainda é bastante desconhecida e subdiagnosticada. O diagnóstico pode ser conseguido por citopatologia ou histopatologia de lesões cutâneas, mas a identificação do agente infeccioso é complexa, uma vez que o crescimento in vitro bacteriano não é possível, dependendo de técnicas moleculares como a PCR para confirmar o DNA de Mycobacterium na amostra. Relatou-se um caso da SGLC em Niterói, estado do Rio de Janeiro, Brasil, diagnosticado por citopatologia e submetido à identificação molecular do agente. Foi realizada amplificação por PCR do gene hsp65, que revelou 100% de homologia genética com a cepa M. murphy. Este é o primeiro relato da SGLC com identificação molecular no estado do Rio de Janeiro, o que mostra a importância de se acrescentar a SGLC ao diagnóstico diferencial das doenças granulomatosas de pele nessa região.(AU)
Subject(s)
Animals , Dogs , Polymerase Chain Reaction/statistics & numerical data , Mycobacterium/cytology , Mycobacterium/pathogenicity , Mycobacterium Infections , DogsABSTRACT
Canine Leproid Granuloma Syndrome (CLGS), also known as canine leprosy, is a cutaneous nodular infectious disease caused by Mycobacterium sp.. Despite being reported worldwide, it is still quite unknown and underdiagnosed. Diagnosis may be achieved by cytopathology or histopathology of skin lesions, but identification of the infectious agent is complex, since bacterial in vitro growth is not possible, relying upon molecular techniques such as PCR to confirm Mycobacterium DNA in the sample. We report a CLGS case in Niteroi, Rio de Janeiro state, Brazil, diagnosed by cytopathology and submitted to molecular identification of the agent. PCR amplification of hsp65 gene was performed and revealed 100% genetic homology to M. murphy strain. This is the first CLGS report with molecular identification in Rio de Janeiro state, and this finding should raise awareness about CLGS as a differential diagnosis among granulomatous skin diseases in this region.(AU)
A síndrome de granuloma leproide canino (SGLC), também conhecida como lepra canina, é uma doença infecciosa cutânea nodular causada por Mycobacterium sp. Apesar de ser relatada mundialmente, ainda é bastante desconhecida e subdiagnosticada. O diagnóstico pode ser conseguido por citopatologia ou histopatologia de lesões cutâneas, mas a identificação do agente infeccioso é complexa, uma vez que o crescimento in vitro bacteriano não é possível, dependendo de técnicas moleculares como a PCR para confirmar o DNA de Mycobacterium na amostra. Relatou-se um caso da SGLC em Niterói, estado do Rio de Janeiro, Brasil, diagnosticado por citopatologia e submetido à identificação molecular do agente. Foi realizada amplificação por PCR do gene hsp65, que revelou 100% de homologia genética com a cepa M. murphy. Este é o primeiro relato da SGLC com identificação molecular no estado do Rio de Janeiro, o que mostra a importância de se acrescentar a SGLC ao diagnóstico diferencial das doenças granulomatosas de pele nessa região.(AU)
Subject(s)
Animals , Dogs , Polymerase Chain Reaction , Mycobacterium/cytology , Mycobacterium/pathogenicity , Mycobacterium Infections , DogsABSTRACT
Pontoporia blainvillei (Gervais and d'Orbigny, 1844) is an endangered small cetacean endemic to South America with four Franciscana Management Areas (FMA) recognized as different population stocks. The role of the intestinal parasite Synthesium pontoporiae (Digenea: Brachycladiidae) as a possible biological marker to differentiate P. blainvillei stocks was evaluated using nuclear and mitochondrial DNA markers. Internal transcribed sequence 1 and 2 (ITS1 and ITS2) regions of S. pontoporiae did not show intraspecific variability. The mitochondrial NADH dehydrogenase subunit 3 (ND3) and cytochrome oxidase subunit I (COI) gene sequences suggested lack of population structure in S. pontoporiae and population expansion. The apparent panmixia of S. pontoporiae may be due to the high mobility of one or more of its intermediary hosts. Alternatively, it may be due to the small sample size. This result is incongruent with the previously proposed FMA.
Subject(s)
Cestode Infections/veterinary , Dolphins/parasitology , Genetic Variation , Intestinal Diseases, Parasitic/veterinary , Platyhelminths/genetics , Platyhelminths/isolation & purification , Animals , Argentina , Brazil , Cestode Infections/epidemiology , Cestode Infections/parasitology , Electron Transport Complex IV/genetics , Endangered Species , Helminth Proteins/genetics , Intestinal Diseases, Parasitic/epidemiology , Intestinal Diseases, Parasitic/parasitology , Molecular Sequence Data , NADH Dehydrogenase/genetics , Phylogeny , Platyhelminths/classification , Platyhelminths/enzymologyABSTRACT
Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized.
Subject(s)
Ecosystem , In Vitro Techniques , Polymerase Chain Reaction/methods , Surface Waters , Vibrio cholerae/genetics , Vibrio cholerae/isolation & purification , Environmental Microbiology , Virulence/genetics , Water SamplesABSTRACT
Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized.
ABSTRACT
Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized.
ABSTRACT
Isolation and genetic characterization of an environmental Vibrio cholerae O1 from the Amazon is reported. This strain lacks two major virulence factors - CTX and TCP - but carries other genes related to virulence. Genetic similarity with epidemic strains is evaluated and the importance of V. cholerae surveillance in the Amazon is emphasized.
ABSTRACT
The objective of the present study was to describe motor behavioral changes in association with histopathological and hematological findings in Wistar rats inoculated intravenously with human T-cell lymphotropic virus type 1 (HTLV-1)-infected MT2 cells. Twenty-five 4-month-old male rats were inoculated with HTLV-1-infected MT2 cells and 13 control rats were inoculated with normal human lymphocytes. The behavior of the rats was observed before and 5, 10, 15, and 20 months after inoculation during a 30-min/rat testing time for 5 consecutive days. During each of 4 periods, a subset of rats was randomly chosen to be sacrificed in order to harvest the spinal cord for histopathological analysis and to obtain blood for serological and molecular studies. Behavioral analyses of the HTLV-1-inoculated rats showed a significant decrease of climbing, walking and freezing, and an increase of scratching, sniffing, biting, licking, and resting/sleeping. Two of the 25 HTLV-1-inoculated rats (8 percent) developed spastic paraparesis as a major behavioral change. The histopathological changes were few and mild, but in some cases there was diffuse lymphocyte infiltration. The minor and major behavioral changes occurred after 10-20 months of evolution. The long-term observation of Wistar rats inoculated with HTLV-1-infected MT2 cells showed major (spastic paraparesis) and minor motor abnormalities in association with the degree of HTLV-1-induced myelopathy.
Subject(s)
Animals , Humans , Male , Rats , Human T-lymphotropic virus 1/physiology , Paraparesis, Tropical Spastic/virology , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Human T-lymphotropic virus 1/genetics , Polymerase Chain Reaction , Paraparesis, Tropical Spastic/blood , Paraparesis, Tropical Spastic/pathology , Time Factors , Viral LoadABSTRACT
The objective of the present study was to describe motor behavioral changes in association with histopathological and hematological findings in Wistar rats inoculated intravenously with human T-cell lymphotropic virus type 1 (HTLV-1)-infected MT2 cells. Twenty-five 4-month-old male rats were inoculated with HTLV-1-infected MT2 cells and 13 control rats were inoculated with normal human lymphocytes. The behavior of the rats was observed before and 5, 10, 15, and 20 months after inoculation during a 30-min/rat testing time for 5 consecutive days. During each of 4 periods, a subset of rats was randomly chosen to be sacrificed in order to harvest the spinal cord for histopathological analysis and to obtain blood for serological and molecular studies. Behavioral analyses of the HTLV-1-inoculated rats showed a significant decrease of climbing, walking and freezing, and an increase of scratching, sniffing, biting, licking, and resting/sleeping. Two of the 25 HTLV-1-inoculated rats (8%) developed spastic paraparesis as a major behavioral change. The histopathological changes were few and mild, but in some cases there was diffuse lymphocyte infiltration. The minor and major behavioral changes occurred after 10-20 months of evolution. The long-term observation of Wistar rats inoculated with HTLV-1-infected MT2 cells showed major (spastic paraparesis) and minor motor abnormalities in association with the degree of HTLV-1-induced myelopathy.
Subject(s)
Human T-lymphotropic virus 1/physiology , Paraparesis, Tropical Spastic/virology , Animals , Disease Models, Animal , Enzyme-Linked Immunosorbent Assay , Human T-lymphotropic virus 1/genetics , Humans , Male , Paraparesis, Tropical Spastic/blood , Paraparesis, Tropical Spastic/pathology , Polymerase Chain Reaction , Rats , Time Factors , Viral LoadABSTRACT
AIM: Rapid characterization of variable region (VR)1 variants of the porA gene among invasive strains is crucial for outbreak management and epidemiology studies. Recent sequence analysis studies in Brazil showed that the VR1 P1.7 and P1.19 variants are highly prevalent, accounting for 68%, of the total number of VR1 variants characterized. The aim of this work is to develop a rapid polymerase chain reaction (PCR)-based method for genosubtyping Neisseria meningitidis by detection of porA variable regions P1.7 and P1.19. METHODS AND RESULTS: PCR primers for the detection of porA VR1 P1.7 and P1.19 were designed and tested using 198 clinical N. meningitidis isolates that had been previously evaluated by porA sequencing. All 50 strains with VR1 P1.7 and all 65 strains with VR1 P1.19 were positively identified by the respective VR-specific PCR and no false-positive reactions occurred. CONCLUSIONS: VR-specific PCR amplification accurately identified VR P1.7 and P1.19 strains. SIGNIFICANCE AND IMPACT OF THE STUDY: To overcome the disadvantages of serosubtyping and sequencing for typing the porA VR1 segment of N. meningitidis, we developed a PCR-based method to rapidly and accurately detect VR1 P1.7 and P1.19 variants. This approach is highly specific and sensitive; moreover it may allow for genotype determination of culture-negative samples.
Subject(s)
Meningococcal Infections/microbiology , Neisseria meningitidis/isolation & purification , Polymerase Chain Reaction , Porins/isolation & purification , Bacterial Typing Techniques , Brazil , DNA Primers , Humans , Meningococcal Infections/diagnosis , Neisseria meningitidis/genetics , Neisseria meningitidis/immunology , Porins/genetics , Porins/immunologyABSTRACT
Previous serological studies on the Arara do Laranjal Indian group revealed extensive HTLV-2 infections. A collection of 97 new samples from the Arara were found repeatedly negative using three different commercial enzyme immunoassays. Eight samples that exhibited optical density readings close to the cut-off value were re-evaluated using Western blot (GeneLab 2.4, Singapore) assay. One sample was found to be non-reactive, five exhibited indeterminate patterns, one was classified as HTLV, and one was confirmed as HTLV-2. Peripheral blood mononuclear cell DNA of the eight samples were subjected to nested PCR and restriction fragment length polymorphism (RFLP) analysis of the pX and env regions, and nucleotide sequencing of the 5'-LTR region. All produced amplification products of pX, but env could be amplified in only one sample with the commonly used primers. RFLP analysis of the pX region using TaqI confirmed HTLV-2 infection. Nucleotide sequencing of the 5'-LTR region was performed in three samples (HTLV-2, HTLV and indeterminate based on Western blot pattern). Phylogenetic analysis of a 449-nt fragment using the Neighbour-Joining method clearly demonstrated that the three samples clustered within the HTLV-2c molecular subtype. The present study confirms the wide dissemination of the HTLV-2c subtype among linguistically and culturally distinct Amazonian Indian groups, and emphasizes the unique occurrence of infection by this subtype in Brazil. Moreover, it emphasizes the limitation of employing the present serological screening assays in blood banks, epidemiological studies, and the importance of molecular assays in the confirmatory procedures for the primary detection of HTLV-2 infections.
Subject(s)
HTLV-I Infections/immunology , HTLV-II Antibodies/blood , Human T-lymphotropic virus 2/isolation & purification , Phylogeny , Brazil/epidemiology , Immunoenzyme Techniques , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , RNA Viruses/isolation & purification , Sensitivity and SpecificityABSTRACT
In the North of Argentina, an endemic area for HTLV-1, intrafamilial transmission of this virus has been observed. The HTLV-1 status in 13 family members of a seropositive blood donor from the central region of Argentina (non-endemic area) was investigated. According to serological and molecular assays, four members of this family (the blood donor, the husband, a son, and a daughter-in-law) proved to be HTLV-1 positive. LTR, tax, and env sequences from the provirus infecting the family members were identical. This strongly suggests the intrafamilial transmission of the virus. This study demonstrated intrafamilial transmission of HTLV-1 in a non-endemic area of Argentina.