ABSTRACT
Background & objectives: Periplasmic copper and zinc superoxide dismutase (Cu,Zn-SOD or SodC) is an important component of the antioxidant shield which protects bacteria from the phagocytic oxidative burst. Cu,Zn-SODs protect Gram-negative bacteria against oxygen damage which have also been shown to contribute to the pathogenicity of these bacterial species. We report the presence of SodC in drug resistant Salmonella sp. isolated from patients suffering from enteric fever. Further sodC was amplified, cloned into Escherichia coli and the nucleotide sequence and amino acid sequence homology were compared with the standard strain sSalmonella Typhimurium 14028. Methods: Salmonella enterica serovar Typhi (S. Typhi) and Salmonella enterica serovar Paratyphi (S. Paratyphi) were isolated and identified from blood samples of the patients. The isolates were screened for the presence of Cu, Zn-SOD by PAGE using KCN as inhibitor of Cu,Zn-SOD. The gene (sodC) was amplified by PCR, cloned and sequenced. The nucleotide and amino acid sequences of sodC were compared using CLUSTAL X. Results: SodC was detected in 35 per cent of the Salmonella isolates. Amplification of the genomic DNA of S. Typhi and S. Paratyphi with sodC specific primers resulted in 519 and 515 bp amplicons respectively. Single mutational difference at position 489 was observed between the sodC of S. Typhi and S. Paratyphi while they differed at 6 positions with the sodC of S. Typhimurium 14028. The SodC amino acid sequences of the two isolates were homologous but 3 amino acid difference was observed with that of standard strain S. Typhimurium 14028. Interpretation & conclusions: The presence of SodC in pathogenic bacteria could be a novel candidate as phylogenetic marker.
Subject(s)
Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Base Sequence , Cloning, Molecular , DNA Primers , Electrophoresis, Polyacrylamide Gel , Molecular Sequence Data , Polymerase Chain Reaction , Salmonella/genetics , Salmonella/metabolism , Sequence Homology, Nucleic AcidABSTRACT
O presente trabalho teve por objetivos determinar possíveis fontes de contaminação dos suínos por Salmonellanas granjas de criação; verificar o comportamento de Salmonellana cadeia produtiva de suínos (do nascimento à terminação e ao longo das etapas de transporte, descanso, abate e frigorificação); correlacionar os sorotipos de Salmonellasp. isolados durante as etapas de criação e pré-abate com aqueles oriundos do abate e frigorificação das carcaças; caracterizar o perfil de resistência de Salmonellasp. utilizando como ferramenta para diferenciação dos sorotipos encontrados, verificar a eficiência de metodologias de isolamento de Salmonella. Desde o nascimento até o abate, a cada 10 dias, foram coletadas amostras de fezes, de ambiente, de ração do cocho, de ração da fábrica e estoque, da água de beber e água de abastecimento em 6 lotes de suínos provenientes de 03 granjas distintas, (2 lotes de cada) localizadas na região oeste do Estado do PR. Destes mesmos lotes, foi acompanhado o pré-abate e abate através da coleta de amostras em 12 pontos ao longo do fluxograma de abate. Durante o confinamento, independente do lote, do ponto ou da data foram isolados 9 sorotipos diferentes de Salmonella(S. 4,5, S. 4,5,12:i:-, S. 4,5,12:i:1,2, S. Agona, S. Derby, S. Mbandaka, S. Panama, S. Poona, S. Typhimurium) e uma cepa não tipável (S. enterica subsp. enterica cepa rugosa). Das amostras coletadas durante o pré-abate e o abate também foram identificadas 9 sorotipos, 3 dos quais não haviam sido isolados nas granjas (S. Give, S. Meleagridis e S. Worthington), sendo que não foram detectados os sorotipos S. Derby, S. 4,5,12:-:1,2 e S. Poona. Foi elevado o percentual de cepas de Salmonellaresistentes à pelo menos um antimicrobiano (93,7%) sendo que 62,9% foram resistentes a mais de um. Pelos resultados encontrados pode-se chegar as seguintes conclusões: a etapa de terminação foi aquela com maior importância para o isolamento de Salmonellasp. durante a criação; a existência de cepas multi-resistentes a antibióticos deve ser considerada como risco à saúde pública; o transporte e descanso dos animais nas pocilgas do matadouro mostraram ser etapas importantes para o aumento da excreção de Salmonellasp. pelos suínos portadores; A contaminação existente nas granjas e em algumas etapas do início do processo de abate não se refletiu nas carcaças abatidas e frigorificadas, em virtude de um processamento tecnológico de abate bem executado; o pré-enriquecimento mostrou ser uma etapa essencial para o isolamento de Salmonellaa partir de fezes e o método alternativo utilizado mostrou ser eficiente como um método de triagem para a identificação de Salmonellasp. das amostras analisadas
The objective of the present trial was to correlate the serotypes of Salmonellasp. isolated during breeding and pre-slaughter with those isolated from slaughter and cooling of carcasses, according to the "from farm to table" concept. From birth to slaughter, every 10 days, samples of feces, the environment of the facility, feed leftovers, feed upon arrival and feed in stock, drinking water and supply water were collected from 6 groups of swine of 3 different breeding units (2 groups of each) located in the west region of the state of Parana, Brazil. The same groups were sampled during pre-slaughter and slaughter in 12 points throughout the slaughter procedure. During confinement, no matter the group, sampling point or date, 9 different Salmonellaserotypes were isolated (S. 4,5, S. 4,5,12:i:-, S. 4,5,12:i:1,2, S. Agona, S. Derby, S. Mbandaka, S. Panama, S. Poona, S. Typhimurium), besides one non-typed strain (rough S. enterica subsp. enterica). From the samples collected during pre-slaughter and slaughter, 9 serotypes were identified, of which 3 had not been isolated in the breeding facilities (S. Give, S. Meleagridis and S. Worthington). Serotypes S. Derby, S. 4,5,12:-:1,2 and S. Poona were not identified. A large percentile of Salmonellastrains was resistant to at least one antimicrobial compound (93.7%), and 62.9% were resistant to more than one. The existence of multiresistant strains should be considered as a public health problem. It was observed that feces presented the greatest epidemiological importance in the dissemination of Salmonellain all breeding stages, and confinement-finishing units were the most important sites of Salmonellasp isolation; transport and rest of the animals in the slaughter pens were important stages in the increase of the Salmonellashedding by carrier animals; there was no direct correlation between the contamination of the breeding facility and positive results obtained in the slaughter, as assessed by carcass contamination
Subject(s)
Salmonella/metabolism , Information Dissemination/methods , Swine/growth & development , Food Contamination/analysis , Foods of Animal Origin , Food Microbiology/instrumentationABSTRACT
Haemolysin patterns of 175 strains of different Salmonella enterica subspecies enterica serovars isolated from different animal sources and places were determined using 11 different blood agar media made with either non-washed horse/sheep erythrocytes or with washed erythrocytes of cattle, sheep, horse, goat, rabbit, guinea pig, and human A, O and B blood groups. Study on 47 strains belonging to 10 serovars of Salmonella from buffalo meat (buffen), 42 strains of 11 serovars from goat meat (chevon): 16 strains of Salmonella enterica serovar Paratyphi B and 25 of S. enterica serovar Paratyphi B var Java from fish, meat, meat products and clinical cases; 45 isolates of S. Abortusequi from aborted mares (18), fetal contents (21), aborted donkey mares (2) and 4 reference strains, revealed that all host restricted Salmonella namely, S. enterica serovar Gallinarum, S. enterica serovar Anatum, S. enterica serovar Abortusequi and S. enterica serovar Paratyphi B could be divided into different haemolysin types based on their inability to produce haemolysis on one or more types of blood agar, while strains of all zoonotic Salmonella serovars induced haemolysis on all the 9 types of blood agar made of washed erythrocytes. None of 175 Salmonella could produce hemolytic colonies on blood agar made of non-washed horse/ sheep erythrocytes. Haemolysin type I (lysing all types of washed erythrocytes) was the commonest one among all serovars except S. Abortusequi, none of which lysed horse erythrocytes. Salmonella enterica serovar Abortusequi having hemolytic activity against sheep erythrocytes were more invasive but had lesser ability to survive in sheep mononuclear cells than non-hemolytic strains. Multiplicity of haemolysins appeared significant epidemiological tool.
Subject(s)
Animals , Buffaloes , Cell Adhesion , Cell Survival , Erythrocytes/microbiology , Hemolysin Proteins/chemistry , Horses , Leukocytes, Mononuclear/metabolism , Mice , Salmonella/metabolism , Salmonella Infections , Salmonella enterica/metabolism , Sheep , Species SpecificityABSTRACT
Eight lactose-fermenting Salmonella agona strains isolated in a pediatric unit were characterized by classic and molecular methods. The strains were classified as biotypes 1a, corresponding to the most frequen one in Brazil. None of the strains produced colicin. Multiple resistence to antimicrobials was observed among the strains studied, It was demonstrated that the lactose-fermenting character was encoded by a plasmid with spontaneous segregaton at a frequency of 1 percer center. This plasmid was transferable by conjugation at a frequency between 4x10(-8) and 5x10(-10). The lac+ plasmid, which molecular weight was approximately 90 MDa, encoded both lactose fermentation and multiple resistance to antimicrobials. Replicon typing showed that this plasmid did not belong to the known types, suggesting the present of a new replicon type. Classic methods showed that the studied strains had the same characteristics as the clone widely occurring in our area, differing only by lactose-fermenting ability. This conclusion was supported by the results of ribotyping study.
Subject(s)
Humans , Infant , Child, Preschool , Salmonella/metabolism , Diarrhea/microbiology , Lactose/metabolism , Drug Resistance, Microbial , Bacterial Typing Techniques , Acute Disease , FermentationABSTRACT
The enterotoxic moiety present in cell free culture supernatant (CFCS) of S. newport strains (MP/120 and BM/704) was purified and antigenically characterised. Purification was achieved to homogeneity by salt precipitation, dialysis and successive gel filtration through Sephadex G-100 and G-200 columns with the help of FPLC set. It was non-dialysable and purified enterotoxin yielded a single protein band in PAGE. It appeared to be of high molecular weight (100 KDa) and highly immunogenic in the rabbit. Antigenically, it was not related to cholera toxin, Shiga toxin or heat-labile enterotoxin of Escherichia coli.
Subject(s)
Antibody Formation , Antigens, Bacterial/immunology , Chromatography, Gel , Enterotoxins/immunology , Salmonella/metabolismABSTRACT
Salmonella weltevreden strains produced a delayed permeability factor (PF) when tested on depilated rabbit skin. The PF activity could be demonstrated in freshly concentrated culture filtrates as well as in the cell lysates. The activity varied with strain and preparation. The induration and blueing reactions were associated with well marked balancing zones.
Subject(s)
Animals , Bacterial Toxins/analysis , Culture Media , Endothelial Growth Factors/analysis , Endotoxins/analysis , India , Lymphokines/analysis , Rabbits , Salmonella/metabolism , Skin/drug effects , Species Specificity , Time Factors , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth FactorsABSTRACT
Investigou-se a produçäo de colicina em 748 amostras de Salmonella (97 sorovares) advindas de díferentes fontes: humana (291), animal (119), ambiental (141), de alimentos (102) e raçöes (95). Detectaram-se 64 amostras (8,6%) colicinogênicas, particularmente isoladas de alimentos (30,4%). ColE1 (53) e Ia (44) foram as mais freqüentes, especialmente no sorovar S, agona, de origem ambiental e de alimentos. Identificou-se também a produçäo de col V em 5 amostras de S. typhimurium dentre 8 culturas produtoras de origem humana. Discute-se a relaçäo entre a capacidade colicinogênica e as fontes e sorovares de Salmonella