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R. bras. Parasitol. Vet. ; 19(4): 256-258, 2010. ilus
Article in English | VETINDEX | ID: vti-4834


This work describes natural infection by Leishmania in a domestic cat where amastigote forms of the parasite were observed in the popliteal lymph node imprint. Positive and negative serological reactions were observed by enzyme linked immunosorbent assay (ELISA) and indirect immunofluorescence assay (IFA), respectively. Polymerase chain reaction (PCR) revealed that the nucleotide sequence of the sample was identical to Leishmania (L.) chagasi. This is the first report of the disease in felines of the city of Andradina, SP, an area considered endemic for canine and human visceral leishmaniasis.(AU)

Neste trabalho, é relatada a infecção natural por Leishmania em um gato doméstico no qual, formas amastigotas do parasito foram observadas em imprint de linfonodo poplíteo. Reações sorológicas positivas e negativas foram observadas pelo teste de imunoadsorção enzimática (ELISA) e reação de imunofluorescência indireta (RIFI), respectivamente. A reação em cadeia da polimerase (PCR) revelou que a sequência de nucleotídeos foi idêntica à Leishmania (L.) chagasi. Este é o primeiro relato da doença em felino da cidade de Andradina, Estado de São Paulo, Brasil, área considerada endêmica para leishmaniose visceral canina e humana.(AU)

Animals , Cats , Leishmania , Leishmaniasis/epidemiology , Cats/parasitology , Polymerase Chain Reaction/methods
Braz. J. Microbiol. ; 46(2): 501-504, Apr.-Jun. 2015. tab, graf
Article in English | VETINDEX | ID: vti-481407


The emergence of β-lactamase-producing Enterobacteriaceae in the last few decades has become major challenge faced by hospitals. In this study, isolates of Klebsiella pneumoniae carbapenemase-2 (KPC-2)-producing K. pneumoniae from a tertiary hospital in Mato Grosso do Sul, Brazil, were characterized. Bacterial identification was performed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF; Bruker Daltonics, Germany) mass spectrometry. The minimum inhibitory concentrations of carbapenems were determined using the agar dilution method as recommended by the Clinical Laboratory Standards Institute guidelines. Carbapenemase production was detected using the modified Hodge test (MHT) and polymerase chain reaction (PCR), followed by DNA sequencing. Of 360 (12.2%) K. pneumoniae isolates obtained between May 2009 and May 2010, 44 (12.2%) were carbapenem nonsusceptible. Of these 44 isolates, thirty-six K. pneumoniae isolates that were positive by MHT and PCR carried the blaKPC-2 gene. Thus, KPC-2producing Klebsiella pneumoniae has been present in a Brazilian hospital located in the Midwest region since at least 2009.(AU)

Humans , Anti-Bacterial Agents/pharmacology , Carbapenems/pharmacology , Klebsiella Infections/microbiology , Klebsiella pneumoniae/enzymology , beta-Lactamases , Brazil , DNA, Bacterial/genetics , Klebsiella pneumoniae/classification , Klebsiella pneumoniae/isolation & purification , Microbial Sensitivity Tests , Polymerase Chain Reaction , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Tertiary Care Centers , beta-Lactamases/genetics
Braz. J. Microbiol. ; 46(4): 1141-1145, Oct.-Dec. 2015. tab
Article in English | VETINDEX | ID: vti-13482


Enterotoxigenic Bacteroides fragilis (ETBF) is an important part of the human and animal intestinal microbiota and is commonly associated with diarrhea. ETBF strains produce an enterotoxin encoded by the bft gene located in the B. fragilis pathogenicity island (BfPAI). Non-enterotoxigenic B. fragilis (NTBF) strains lack the BfPAI and usually show two different genetic patterns, II and III, based on the absence or presence of a BfPAI-flanking region, respectively. The incidence of ETBF and NTBF strains in fecal samples isolated from children without acute diarrhea or any other intestinal disorders was determined. All 84 fecal samples evaluated were B. fragilis-positive by PCR, four of them harbored the bft gene, 27 contained the NTBF pattern III DNA sequence, and 52 were considered to be NTBF pattern II samples. One sample was positive for both ETBF and NTBF pattern III DNA sequences. All 19 B. fragilis strains isolated by the culture method were bft-negative, 9 belonged to pattern III and 10 to pattern II. We present an updated overview of the ETBF and NTBF incidence in the fecal microbiota of children from Sao Paulo City, Brazil.(AU)

Humans , Child , Bacteroides fragilis , Feces , Firmicutes , Polymerase Chain Reaction
J Zoo Wildl Med ; 52(4): 1159-1166, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34998285


Leprosy has been described in Eurasian red squirrel (Sciurus vulgaris; ERS) carcasses since 2014. Studies of ERS carcasses have not provided information about incubation or disease progression in this host but have provided important insights into pathogen presence and distribution throughout the United Kingdom. Here we present field study data on 31 live ERS from an island population naturally infected with Mycobacterium leprae that were assessed longitudinally over a 2-yr time period. Clinical assessment, serologic (anti-phenolic glycolipid-I antibody [αPGL-I] detection) and molecular methods (polymerase chain reaction) were used to diagnose and categorize ERS at each assessment as a leprosy case, a leprosy suspect, colonized by M. leprae, or a contact ERS. Eight ERS (25.8%) were identified as leprosy cases: four at initial assessment, two at 6 mon and two at 24 mon after initial assessment. One ERS was categorized a leprosy suspect when it developed typical lesions 12 mon after initial assessment, despite negative serologic and molecular test results at this time, though M. leprae DNA had been isolated during the initial assessment. Seven ERS (22.6%) were categorized as colonized and of these, six were reassessed but did not develop clinical signs of leprosy within 6 (n = 2), 12 (n = 3), and 18 (n = 1) mon. Most (48.4%, n = 15) were categorized as contact ERS. Progression of leprosy lesions varied between ERS, but always increased in severity over time and was paralleled with increased antibody response. Based on our dataset, we propose the hypotheses: 1) leprosy in ERS is a chronic, slowly progressing disease in this species, similar to that described for other hosts; 2) lesions can undergo repeated ulceration-healing cycles; and 3) in some instances M. leprae DNA and αPGL-I antibodies are detectable before the onset of clinical signs of disease. Future studies addressing the progression of leprosy in ERS should follow affected animals over a longer time period and include tissue samples to pair molecular diagnostics with serologic results.

Leprosy , Rodent Diseases , Animals , Antibodies , Leprosy/diagnosis , Leprosy/epidemiology , Leprosy/veterinary , Mycobacterium leprae , Polymerase Chain Reaction/veterinary , Sciuridae
J Zoo Wildl Med ; 52(4): 1275-1279, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34998300


Herpesviruses are important pathogens in zoologic chelonian collections and have been associated with fatal disease in turtles of the family Emydidae. In this report, three western pond turtles (Actinemys marmorata), living in a mixed-species freshwater turtle exhibit, presented with varying degrees of lethargy, pallor, generalized edema, and cloacal hemorrhage before death within a 2-wk period. Postmortem findings included necrohemorrhagic enterocolitis, necrotizing splenitis, hepatic necrosis, esophagitis, thymic necrosis, and pneumonia with epithelial necrosis and degeneration of the trachea and kidneys. Intraepithelial, intranuclear, amphophilic to eosinophilic, Cowdry type A viral inclusion bodies were identified in the intestinal tract, liver, spleen, kidney, trachea, lung, and thymus. PCR amplification and sequencing of liver tissue produced amplicons that were 100% homologous with emydid herpesvirus 1 (EmyHV-1). Molecular screening of cohoused emydid turtles revealed that a red-eared slider (Trachemys scripta elegans) and a western pond turtle, both asymptomatic, were PCR-positive for EmyHV-1 on combined oral-cloacal swabs. This report describes, for the first time, EmyHV-1-associated disease in western pond turtles and molecularly identifies EmyHV-1 in an asymptomatic red-eared slider.

Alphaherpesvirinae , Herpesviridae Infections/veterinary , Herpesviridae , Turtles , Animals , Animals, Zoo , Herpesviridae Infections/epidemiology , Polymerase Chain Reaction/veterinary
J Zoo Wildl Med ; 52(4): 1309-1313, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34998304


A 4-yr-old male intact lesser spot-nosed guenon (Cercopithecus petaurista), housed at a North American zoological facility, presented with acute lethargy, inappetence, and mild neurologic signs. Physical examination revealed hemorrhagic pleural effusion in the right hemithorax. This guenon's condition improved over several days but then deteriorated, and the guenon presented with lethargy and weakness. A hemorrhagic pleural effusion was identified within the left hemithorax. The guenon developed respiratory and cardiac arrest while anesthetized. Gross examination revealed tract formation in the liver, adhesions of the liver to the diaphragm, hemorrhagic thoracic and abdominal effusion, and a single trematode within the right hemithorax. Morphologic features and species identification by PCR confirmed that the parasite was Fascioloides magna. Histologic examination revealed tract formation in the liver associated with biliary hyperplasia, fibrosis and hepatic necrosis, severe bile peritonitis, and pleuritis. This is the first report of an infection by F. magna in a primate.

Cercopithecus , Fasciolidae , Trematode Infections/veterinary , Animals , Animals, Zoo , Cercopithecus/parasitology , Fasciolidae/genetics , Fatal Outcome , Liver , Male , Polymerase Chain Reaction/veterinary
BMC Genomics ; 23(1): 31, 2022 Jan 06.
Article in English | MEDLINE | ID: mdl-34991471


BACKGROUND: Short read sequencing has been used extensively to decipher the genome diversity of human cytomegalovirus (HCMV) strains, but falls short to reveal individual genomes in mixed HCMV strain populations. Novel third-generation sequencing platforms offer an extended read length and promise to resolve how distant polymorphic sites along individual genomes are linked. In the present study, we established a long amplicon PacBio sequencing workflow to identify the absolute and relative quantities of unique HCMV haplotypes spanning over multiple hypervariable sites in mixtures. Initial validation of this approach was performed with defined HCMV DNA templates derived from cell-culture enriched viruses and was further tested for its suitability on patient samples carrying mixed HCMV infections. RESULTS: Total substitution and indel error rate of mapped reads ranged from 0.17 to 0.43% depending on the stringency of quality trimming. Artificial HCMV DNA mixtures were correctly determined down to 1% abundance of the minor DNA source when the total HCMV DNA input was 4 × 104 copies/ml. PCR products of up to 7.7 kb and a GC content < 55% were efficiently generated when DNA was directly isolated from patient samples. In a single sample, up to three distinct haplotypes were identified showing varying relative frequencies. Alignments of distinct haplotype sequences within patient samples showed uneven distribution of sequence diversity, interspersed by long identical stretches. Moreover, diversity estimation at single polymorphic regions as assessed by short amplicon sequencing may markedly underestimate the overall diversity of mixed haplotype populations. CONCLUSIONS: Quantitative haplotype determination by long amplicon sequencing provides a novel approach for HCMV strain characterisation in mixed infected samples which can be scaled up to cover the majority of the genome by multi-amplicon panels. This will substantially improve our understanding of intra-host HCMV strain diversity and its dynamic behaviour.

Cytomegalovirus , High-Throughput Nucleotide Sequencing , Cytomegalovirus/genetics , Haplotypes , Humans , Polymerase Chain Reaction , Sequence Analysis, DNA