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Siderastrea stellata is a coral endemic to Brazilian waters and is widely distributed throughout the coast of Bahia state. Exposure of the coral to pathogens stimulates the production of important enzymes by associated symbiont microorganisms. Besides being natural sources of bioactive compounds, these microorganisms possess characteristics that help them survive under extreme conditions. Sixty-eight bacterial strains isolated from the S. stellata coral were analyzed for antimicrobial activity against ten pathogenic bacteria. The cultivation of marine isolates was carried out in liquid or solid seawater. Eight isolates produced antimicrobial compounds against at least two of the ten pathogenic bacteria tested. When isolates were grown in Mueller-Hinton broth, a distinct biocidal spectrum was observed, indicating that the culture medium directly influences the production of antimicrobial compounds. Through molecular characterization of activity-positive isolates, they were identified as belonging to the genus Bacillus . The bioactive compounds produced by the Bacillus stratosphericus SS85 and SS69 strains were stable after storage time for three months. This is the first paper reporting on the antimicrobial activity of microorganisms isolated from coral Siderastrea stellata .

We aimed to measure differences in hindgut pH, intestinal histological parameters and occurrence of enteric pathogens in the collared peccary (Dicotyles tajacu), according to dietary fiber levels and interindividual behavioral differences. Samples of ileum and cecum (tissue and content) were collected from 24 adult males classified according to their behavioral traits, and which received either low (n = 12) or high-fiber (n = 12) diets. Salmonella spp. was discovered in the cecum content of the most distressed individual. Non-pathogenic E. coli was present in 21 of the 24 collared peccaries. The pH in cecum samples was higher when animals were fed with high-fiber diet. The depth of the crypt (CD) was correlated with the peccaries’ behavioral traits: the more relaxed, the higher the CD. Associations linking individual behavioral qualities, dietary fiber, pH, histology of the hindgut and the presence of enteric pathogens in farmed peccaries are multifaceted. The existence of Salmonella in the hindgut of the distressed individual and the positive impact of behavioral calmness traits on some intestinal histological parameters suggests the possible benefit of selecting calm individuals, from the point of view of both animal and human health and welfare.

Wild animals have an ecological function and can serve as sentinels to identify infectious agents and as indicators of environmental health. Among the zoonotic pathogens, Salmonella spp. deserve special attention due to their high worldwide prevalence and their ubiquity of hosts. With the aim of investigating the presence of Salmonella spp. in wild birds from the Atlantic Forest in southern Bahia, Brazil, we collected 114 fecal samples of wild birds (14 families) between 2016 and 2017. Fecal samples were collected by means of cloacal swab and subjected to microbiological culture to isolate and serotype Salmonella spp. specifically. Antibiotic susceptibility was determined using the disk diffusion test protocol. Only one bird, Ceratopipra rubrocapilla, tested positive for Salmonella enterica subsp. enterica serotype Agona, which is the first record for this bird species. This isolate exhibited intermediate sensitivity to amikacin and gentamicin and sensitivity to the other 13 antibiotics tested. Results may indicate environmental preservation since the studied areas had minimal human activity and good sanitary quality. Despite the low prevalence, it is necessary to monitor wildlife and establish disease control and surveillance systems, especially for zoonotic diseases.

Microorganisms colonize the corals and produce compounds with interesting biological properties for biotechnology. The aim of the present study was to search for biocompounds with industrial potential through the cloning metagenomic DNA of symbionts the coral S. stellata in fosmid vector. The metagenomic library was analyzed using functional screening and sequencing via the Illumina MiSeq. Of the 3648 clones, eight were identified as proteolytic and six as amylolytic. The proteolytic clone, P07H3, also exhibited antimicrobial activity against S. aureus, S Enterica PT11, S Enterica PT4 and S Typhi. No clones were positive for lipase/esterase. Beta-lactamase was a single subsystem identified by Rapid Annotation using Subsystem Technology (RAST) through fosmid sequencing of clone P07H3. However, there were identifications of enzymes that participate in metabolic processes such as polysaccharide catabolism, oxidation-reduction, signal transduction and phosphorylation. When the hypothetical proteins were re-analyzed they exhibited a defined functional domain. All the identified open reading frames (ORFs) had low identity with proteins deposited in BLASTp. The sequencing shows that most of the genomic fragments of clone P07H3 possibly consist of new proteins. The Proteobacteria phylum had the greatest predominance among the analyzed fragments. This is the first report on the prospection of amylases, proteases and antimicrobials of coral S. stellata samples. The analyzes of this study help to improve the knowledge about the metabolic diversity of S. stellata still little explored and viable in the future study of the identified compounds.

This study evaluated the effect of essential oil (ESOL) of Mentha piperita and Melaleuca alternifolia on meat-type quails. To examine performance, a completely randomized design was used, with four treatments and eight repetitions each, using seven birds per repetition. To assess lipidemia, a completely randomized design in a 4 × 2 factorial scheme was used, with four treatments and two collection conditions using eight repetitions and one bird per repetition. Weight gain, feed intake, feed conversion, carcass yield, cholesterol, high-density lipoporteins, triglycerides, pH, and brightness of quail breast meat were assessed. The data were analyzed for homoscedasticity and normality, and the means were subjected to analysis of variance. Adding M. piperita and M. alternifolia ESOL to the diets of meat-type quails can improve performance, carcass yield, and meat quality characteristics, comparable to the use of performance-enhancing antibiotics. The use of zinc bacitracin, M. piperita, and M. alternifolia under different collection conditions did not increase the serum levels of low-density and very low-density lipoproteins. The variation coefficients measured after 12 hours of fasting were 86% lower than without fasting.

The sharp increase of COVID-19 cases in late 2020 has made Brazil the new epicenter of the ongoing SARS-CoV-2 pandemic. The novel viral lineages P.1 (Variant of Concern Gamma) and P.2, respectively identified in the Brazilian states of Amazonas and Rio de Janeiro, have been associated with potentially higher transmission rates and antibody neutralization escape. In this study, we performed the whole-genome sequencing of 185 samples isolated from three out of the five Brazilian regions, including Amazonas (North region), Rio Grande do Norte, Paraíba and Bahia (Northeast region), and Rio de Janeiro (Southeast region) in order to monitor the spread of SARS-CoV-2 lineages in Brazil in the first months of 2021. Here, we showed a widespread dispersal of P.1 and P.2 across Brazilian regions and, except for Amazonas, P.2 was the predominant lineage identified in the sampled states. We estimated the origin of P.2 lineage to have happened in February, 2020 and identified that it has differentiated into new clades. Interstate transmission of P.2 was detected since March, but reached its peak in December, 2020 and January, 2021. Transmission of P.1 was also high in December and its origin was inferred to have happened in August 2020. We also confirmed the presence of lineage P.7, recently described in the southernmost region of Brazil, to have spread across the Northeastern states. P.1, P.2 and P.7 are descended from the ancient B.1.1.28 strain, which co-dominated the first phase of the pandemic in Brazil with the B.1.1.33 strain. We also identified the occurrence of a new lineage descending from B.1.1.33 that convergently carries the E484K mutation, N.9. Indeed, the recurrent report of many novel SARS-CoV-2 genetic variants in Brazil could be due to the absence of effective control measures resulting in high SARS-CoV2 transmission rates. Altogether, our findings provided a landscape of the critical state of SARS-CoV-2 across Brazil and confirm the need to sustain continuous sequencing of the SARS-CoV-2 isolates worldwide in order to identify novel variants of interest and monitor for vaccine effectiveness., Author summary Since its first detection in December 2019, the SARS-CoV-2 has evolved into more than a thousand recognized lineages. Several of these lineages are known to have higher transmissibility or better escape from the immune system. One of them is the P.1 lineage, also known as the variant of concern Gamma. It was first discovered in Brazil in December 2020 and has quickly replaced the previous lineage dominating the country, P.2. We used genomic data from SARS-CoV-2 samples that were collected in the first months of 2021 to analyze how P.1, P.2 and other lineages had spread across Brazil. Our research has identified that P.1 lineage was already present in several states of Brazil almost two months before its first detection in the state of Amazonas. Our work sheds light on the importance of continuous monitoring of SARS-CoV-2 lineages in historically understudied regions to early detect and control the spread of new variants of concern.

An 11‐year‐old male mixed‐breed cat, with exclusively indoor life, presented 3 cough episodes after the owners tested positive by RT‐PCR for SARS‐CoV‐2. The house is inhabited by 5 people (3 adults and 2 children), and 2 of the adults have shown mild symptoms associated with throat discomfort. The cat was vaccinated, had no history of any previous disease, and tested negative for feline coronavirus (FCoV), feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV). Rectal sample collected from the cat was positive for SARS‐CoV‐2 by RT‐PCR. Viral genome sequences recovered from human and cat samples showed an average 99.4% sequence identity. This is the first report of genome sequences of SARS‐CoV‐2 recovered from a cat and its owner in Latin America.

The genotyping of 25 isolates of Toxoplasma gondii from free-range chickens in the state of Bahia, Brazil, was performed by PCR-restriction fragment length polymorphism using 11 genetic markers: SAG1, 5′+3′SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico, and CS3. The analysis revealed 8 genotypes, 3 of which had not been previously reported. Four genotypes were represented by single isolates, whereas the other genotypes were represented by 2 or more isolates. Five isolates showed mixed infections, and 2 of them were identical. None of the clonal types I, II, or III were found, but 2 isolates corresponded to the Brazilian clonal lineage BrIII. There was a single allele for the c22-8 marker. The CS3 marker demonstrated efficiency in the evaluation of virulence in mice. This study reaffirms the diverse genetic variability of T. gondii in Brazil.

Strains of diarrheagenic Escherichia coli (DEC) are involved in foodborne disease outbreaks worldwide, especially the enterohemorrhagic E. coli O157:H7. This study describes two multiplex quantitative real time PCR (qPCR) assays for simultaneous identification and quantification of genes related to virulence of DEC; a triplex reaction for detection and quantification of stxA1, stxA2, and eaeA genes, and a duplex reaction for detection and quantification of eaeA and virA genes. The technique was applied in raw oyster samples for direct quantification of DEC, thereby evaluating the applicability of this methodology for microbiological quality assessment of food. Using custom designed primers and specific MGB probes, a triplex qPCR assay was performed to quantify stxA1, stxA2, and eaeA, and a duplex reaction was performed to quantify virA and eaeA genes. The assays showed high sensitivity, with the detection limit varying between 5 and 17 copies of the genes. The coefficient of determination (R2) of the standard curves was 0.99. The coefficient of variation was

Strains of Toxoplasma gondii in Brazil are highly genetically diverse compared to strains from North America and Europe. Dogs are epidemiologically important because they act as sentinels for T. gondii infections in humans and are good indicators of environmental contamination. The aim of this study was to isolate and genetically characterize T. gondii strains from tissues of naturally infected Brazilian dogs. For this study, 21 blood samples were collected from dogs at the Zoonosis Control Centers of Ilheus and Itabuna cities, Bahia, Brazil. The sera were examined for T. gondii antibodies using the indirect hemagglutination test. Brains and hearts of seropositive dogs were bioassayed in mice to isolate and characterize T. gondii parasites by PCR-RFLP using 10 genetic markers (SAG1, newSAG2, SAG3, BTUB, c22-8, c29-2, GRA6, PK1, APICO, and L358). However, T. gondii was isolated from only 4 (57.1%) dogs, designated TgDgBr6, 13, 17, and 21. All strains were virulent, causing clinical changes (rough hair coat, lethargy, and abdominal distention) and the death of all mice within 8-20 days after inoculation. Genetic analysis of these 4 T. gondii isolates revealed 4 distinct genotypes with different clonal lineage combinations (types I, II, and III) and 2 atypical alleles. Using PCR-RFLP with several markers, this study contributes to evaluations of the genetic diversity of strains circulating in Brazil.

Quantitative real-time polymerase chain reactions (qPCRs) of the most prevalent bacteria causing foodborne diseases worldwide, such asSalmonellaspp.,Escherichia coli, andStaphylococcus aureus,can be an important tool for quantitative microbial risk assessment, which requires numerical data to determine the level of contamination at a specific stage of food production. However, most of qPCR assays described in the literature for these pathogens are qualitative; their objective is pathogen detection and not pathogen quantification. Thus, the aim of our work was to develop a qPCR for the simultaneous quantification ofSalmonellaspp.,E. coli, andS. aureusand to propose its use in the analysis of foods, as a tool for microbiological quality monitoring. For this, a multiplex qPCR was standardized for the simultaneous quantification of specific fragments of target genes (ssf,phoA, andnuc) corresponding to each one of the mentioned bacteria. The limit of detection of the technique was 13, 10, and 12 gene copies forssf,phoA, andnuc, respectively; standard curves showed R2> 0.99, with efficiencies ranging from 99 to 110%, and inter- and intraexperiment reproducibility presented a low coefficient of variation in all trials. This methodology was applied in different food matrices (milk, ground beef, and oyster meat), and the results were compared with official microbiological culture methodology and with ready-to-use test. Advantages and disadvantages of each methodology used in this study are pointed out. We suggest that this multiplex qPCR can be used as a rapid screening technique for the analysis of food microbiological quality.

Strains of diarrheagenic Escherichia coli (DEC) are involved in foodborne disease outbreaks worldwide, especially the enterohemorrhagic E. coli O157:H7. This study describes two multiplex quantitative real time PCR (qPCR) assays for simultaneous identification and quantification of genes related to virulence of DEC; a triplex reaction for detection and quantification of stxA1, stxA2, and eaeA genes, and a duplex reaction for detection and quantification of eaeA and virA genes. The technique was applied in raw oyster samples for direct quantification of DEC, thereby evaluating the applicability of this methodology for microbiological quality assessment of food. Using custom designed primers and specific MGB probes, a triplex qPCR assay was performed to quantify stxA1, stxA2, and eaeA, and a duplex reaction was performed to quantify virA and eaeA genes. The assays showed high sensitivity, with the detection limit varying between 5 and 17 copies of the genes. The coefficient of determination (R(2)) of the standard curves was 0.99. The coefficient of variation was

The objective of this study was to standardize the high-resolution melting method for identification and discrimination of Toxoplasma gondii, Sarcocystis spp., Neospora spp., and Cryptosporidium spp. by amplification of 18S ribosomal DNA (rDNA) using a single primer pair. The analyses were performed on individual reactions (containing DNA from a single species of a protozoan), on duplex reactions (containing DNA from two species of protozoa in each reaction), and on a multiplex reaction (containing DNA of four parasites in a single reaction). The proposed method allowed us to identify and discriminate the four species by analyzing the derivative, normalized, and difference melting curves, with high reproducibility among and within the experiments, as demonstrated by low coefficients of variation (less than 2.2% and 2.0%, respectively). This is the first study where this method is used for discrimination of these four species of protozoa in a single reaction.

The microbiological quality of oysters reflects the microbiological quality of their habitats because they are filter feeders. The objective of this study was to assess the bacterial composition of the edible oyster Crassostrea rhizophorae in urban and preserved estuaries. Particularly, we assessed the presence of pathogenic bacteria, investigated antibiotic susceptibility in bacterial isolates, and quantified β-lactam antibiotic resistance genes (blaTEM, blaSHV, and blaKPC) via quantitative PCR of oyster DNA. Our results detected total coliforms, Escherichia coli, and enterobacteria in the oysters from urban estuaries, which is indicative of poor water quality. In addition, our detection of the eaeA and stxA2 virulence genes in 16.7% of E. coli isolates from oysters from this region suggests the presence of multiantibiotic-resistant enteropathogenic and enterohemorrhagic E. coli strains. During periods of low precipitation, increased contamination by E. coli (in winter) and Vibrio parahaemolytic...

This work reports the distribution of an oral dose of Salmonella enterica serovar Enteritidis (SE) in C57Bl/6-Bcgr mice, to study its pathogenesis in a latent carrier animal. Mice orally inoculated with a high dose of SE developed a latent infection characterized by the absence of clinical symptoms in which the cecum is functioning as a “strategic site” of SE proliferation, releasing bacteria into feces intermittently over the 4-week study. A sequence of disruptions occurred in the small intestine at 1 day postinculation (PI). The microvilli exhibited different degrees of degeneration, which were reversible as the cells became vacuolated. From 2 days PI, SE was detected in the mononuclear phagocytic system, and an exponential growth of the remaining bacteria in tissues was observed until 4 days PI. The production of interferon gamma from 3 days PI is restricting the SE growth, and a plateau phase was observed from 4 to 15 days PI. A recurrence of the bacterial growth in tissue occurred from 15 to 28 days PI, especially in the cecum. Increasing our knowledge about the host–pathogen interaction of adapted pathogens with the ability to develop latency is essential for the development of an efficient strategy for Salmonella control.

Strains of Toxoplasma gondii in Brazil are highly genetically diverse compared to strains from North America and Europe. Dogs are epidemiologically important because they act as sentinels for T. gondii infections in humans and are good indicators of environmental contamination. The aim of this study was to isolate and genetically characterize T. gondii strains from tissues of naturally infected Brazilian dogs. For this study, 21 blood samples were collected from dogs at the Zoonosis Control Centers of Ilhéus and Itabuna cities, Bahia, Brazil. The sera were examined for T. gondii antibodies using the indirect hemagglutination test. Brains and hearts of seropositive dogs were bioassayed in mice to isolate and characterize T. gondii parasites by PCR-RFLP using 10 genetic markers (SAG1, newSAG2, SAG3, BTUB, c22-8, c29-2, GRA6, PK1, APICO, and L358). However, T. gondii was isolated from only 4 (57.1%) dogs, designated TgDgBr6, 13, 17, and 21. All strains were virulent, causing clinical changes (rough hair coat, lethargy, and abdominal distention) and the death of all mice within 8–20 days after inoculation. Genetic analysis of these 4 T. gondii isolates revealed 4 distinct genotypes with different clonal lineage combinations (types I, II, and III) and 2 atypical alleles. Using PCR-RFLP with several markers, this study contributes to evaluations of the genetic diversity of strains circulating in Brazil.

This study genetically Toxoplasma gondii isolates obtained from pigs intended for human consumption in northeastern Brazil; multilocus PCR-RFLP and sequencing techniques were utilized. Bioassays were conducted using the brain and tongue of 20 pig heads purchased at butcher shops in the city of Ilheus, Bahia, Brazil. Overall, 11 T. gondii isolates designated TgPgBr06-16 were identified. Application of multilocus PCR-RFLP with seven molecular markers (SAG1, SAG2, SAG3, BTUB, C22-8, PK1 and Apico) identified six different genotypes. Isolates TgPgBr 06, 08, 11, 12, 14 and 15 were indistinguishable by this technique, forming a single genotype; the remaining isolates were characterized as distinct genotypes. However, when five genetic markers (SAG1, SAG2, SAG3, BTUB and c22-8) were employed in multilocus PCR-sequencing, all eleven strains of T. gondii were shown to be different. All isolates differed from Type I, II and III clonal genotypes using both genotyping techniques. These results demonstrate that the multilocus PCR-RFLP assay underestimated the true diversity of the T. gondii population in this study. Thus, DNA sequencing is the preferred technique to infer the genetic diversity and population structure of T. gondii strains from Brazil. Moreover, it is necessary to develop new molecular markers to group and characterize atypical T. gondii isolates from South America.

Species of tegu (Tupinambis) are the largest lizards in South America. Large numbers of these lizards are hunted; there is a vigorous trade in their skins and the meat is consumed by rural and native peoples. The animals are also bred in captivity, an economic activity for rural populations which can help in the animals' conservation. Faecal samples from 30 captive-born tegus were analysed for the presence of Salmonella in two separate samplings. In the first analysis, samples from 26 animals (87%) yielded Salmonella enterica of which 23% were of Rubislaw serotype; 20% Carrau and Agona serotypes; 7% Infantis and Saint-Paul serotypes; 3% Panama and Brandenburg serotypes; 10% were S. enterica subsp. enterica and 7% were rough form. In the second analysis, four tegus (13%) which had been negative in the first sampling were positive, thus, 100% of the animals studied carried the bacterium. Antibiotic susceptibility showed resistance to sulfonamide in 82% of the isolates, streptomycin in 64%, tetracycline in 6% and Chloramphenicol in 20%. Two animals carried strains of the same serotype with different patterns of antibiotic susceptibility. Although it is well known that reptiles are a significant source of Salmonella, to our knowledge, its prevalence in tegu has not been studied previously.

Landfarm soil is used to bioremediate oil wastes from petrochemical industries. We developed a simplified protocol for microbial DNA extraction of tropical landfarm soil using only direct lysis of macerated material. Two samples of tropical landfarm soil from a Brazilian refinery were analyzed by this protocol (one consisted of crude oil-contaminated soil; the other was continuously enriched for nine months with petroleum). The soil samples were lysed by maceration with liquid nitrogen, eliminating the need for detergents, organic solvents and enzymatic cell lysis. Then, the DNA from the lysed soil sample was extracted using phenol-chloroform-isoamyl alcohol or guanidium isothiocyanate, giving high DNA yields (more than 1 micro g DNA/g soil) from both soil types. This protocol compared favorably with an established method of DNA template preparation that included mechanical, chemical and enzymatic treatment for cell lysis. The efficiency of this extraction protocol was confirmed by polymerase chain reaction amplification of the 16S rRNA gene, denaturing gradient gel electrophoresis and cloning assays. Fifty-one different clones were obtained; their sequences were classified into at least seven different phyla of the Eubacteria group (Proteobacteria - alpha, gamma and delta, Chloroflexi, Actinobacteria, Acidobac teria, Planctomycetes, Bacteroidetes, and Firmicutes). Forty percent of the sequences could not be classified into these phyla, demonstrating the genetic diversity of this microbial community. Only eight isolates had sequences similar to known sequences of 16S rRNA of cultivable organisms or of known environmental isolates and therefore could be identified to the genus level. This method of DNA extraction is a useful tool for analysis of the bacteria responsible for petroleum degradation in contaminated environments.

The objective of this study was to verify whether Toxoplasma gondii is excreted in the milk of naturally infected sheep. In order to accomplish this, 275 lactating ewes were used; these were bred extensively in 17 estates distributed across nine cities. Polymerase chain reaction amplification was used to detect T. gondii DNA in milk samples, and the indirect immunofluorescence test was employed for the detection of anti-T. gondii IgG antibodies in the sera, with a cut-off value of 1:64. It was possible to verify the presence of the parasite DNA in 6.5% (18/275) of the studied animals. Anti-T. gondii antibodies were present in 41.5% of the animals studied (114/275). There was no correlation between parasite excretion in milk and the presence of IgG in 38.9% of the studied animals (7/18). The high seropositivity and the presence of parasite DNA in the milk led to the conclusion that T. gondii infection is present in the sheep population in southern and southwestern Bahia, and that there is a risk of the human population becoming infected due to the consumption of raw, in natura milk.

The aim of this study was to detect evidence of Toxoplasma gondii using polymerase chain reaction (PCR)-based techniques in oysters (Crassostrea rhizophorae) obtained from the southern coastal region of Bahia, Brazil. A total of 624 oysters were collected, and the gills and digestive glands were dissected. Each tissue sample was separated into pools containing tissues (of the same type) from three animals, leading to a total of 416 experimental samples for analysis (208 samples each from the gills and digestive glands). Molecular analysis using PCR-based detection of the T. gondii AF 146527 repetitive fragment yielded negative results for all samples. However, when nested-PCR was used for detection of the T. gondii SAG-1 gene, 17 samples were positive, with the gills being the tissue with maximal detection of the parasite. These positive results were confirmed by sample sequencing. It is therefore suggested that C. rhizophorae oysters are capable of filtering and retaining T. gondii oocysts in their tissue. This represents a risk to public health because they are traditionally ingested in natura.

Background Strains of Toxoplasma gondii in Brazil have high genetic diversity compared to North America and Europe. The bristle-spined porcupine, Chaetomys subspinosus, is often subject to hunting for human food, but it is not known whether it can be a reservoir of this parasite. The aim of this study was to verify the occurrence of T. gondii in C. subspinosus from southern Bahia, Brazil, and genetically characterize and compare the strains found with those isolated in previous studies of the same region to quantify their genetic diversity by multilocus PCR-RFLP and PCR sequencing. Findings Twelve free-ranging C. subspinosus captured in forest fragments of the Una Biological Reserve and adjacent areas were evaluated. Three isolates of T. gondii (TgCsBr01-03) were detected. Two different genotypes were identified by applying multilocus PCR-RFLP with six molecular markers (SAG1, SAG2, SAG3, c22-8, PK1, and Apico). The isolates TgCsBr02 and TgCsBr03 were indistinguishable by this technique. However, the three isolates differed from all the reference strains and from the samples from the same region. Nevertheless, when the six genetic markers were used in multilocus PCR sequencing, all three isolates of T. gondii were different. The phylogenetic analysis revealed a greater genetic distance for TgCsBr01, which was closer to isolates from pigs from the same region, while TgCsBr02-03 was classified in the same lineage and was closer to isolates from sheep from this region. Conclusions All the isolates differed from the clonal genotypes of types I, II, and III using both genotyping techniques.

Recent studies have demonstrated that strains of Toxoplasma gondii in Brazil are frequently different from those detected in other countries, thus making an accurate phylogenetic analysis difficult. The aim of this study was to genetically characterize T. gondii samples from sheep raised in southern Bahia and intended for human consumption, by means of PCR–RFLP and sequencing techniques. Experimental samples were obtained from 200 sheep brains purchased at butcher's shops in Itabuna, Bahia, Brazil. In total, three samples (#54, #124 and #127) were T. gondii-positive. The application of multilocus PCR–RFLP using ten molecular markers (SAG1, SAG2, SAG3, BTUB, c22-8, PK1, GRA6, L358, c-29-2 and Apico) revealed a single genotype common to all samples of this study, which differed from any other published T. gondii genotypes. An atypical allele was detected in the L358 genetic marker; this has not previously been shown in any other South American T. gondii isolates. Phylogenetic analysis on the sequences from multilocus PCR sequencing revealed that these three samples were classified into the same lineage. Extensive indel regions were detected in the Apico genetic marker. Together, our findings revealed a new Brazilian T. gondii genotype. Further research should be conducted to enrich the database of Brazilian T. gondii genotypes from different regions. This will make it possible to understand the phylogenetic relationship between isolates.

This work reports the distribution of an oral dose of Salmonella enterica serovar Enteritidis (SE) in C57Bl/6-Bcgr mice, to study its pathogenesis in a latent carrier animal. Mice orally inoculated with a high dose of SE developed a latent infection characterized by the absence of clinical symptoms in which the cecum is functioning as a "strategic site" of SE proliferation, releasing bacteria into feces intermittently over the 4-week study. A sequence of disruptions occurred in the small intestine at 1 day postinculation (PI). The microvilli exhibited different degrees of degeneration, which were reversible as the cells became vacuolated. From 2 days PI, SE was detected in the mononuclear phagocytic system, and an exponential growth of the remaining bacteria in tissues was observed until 4 days PI. The production of interferon gamma from 3 days PI is restricting the SE growth, and a plateau phase was observed from 4 to 15 days PI. A recurrence of the bacterial growth in tissue occurred from 15 to 28 days PI, especially in the cecum. Increasing our knowledge about the host-pathogen interaction of adapted pathogens with the ability to develop latency is essential for the development of an efficient strategy for Salmonella control.

The oil and petrochemical industry in the course of the exploration, production, refining and transportation operations required for the commercialization of oil and its derivatives have contaminated with toxic wastes the environment, which requires urgent remediation. The biological treatment of these wastes aims the rational exploitation of microorganisms that use hydrocarbons as the source of carbon and energy. This biotechnological strategy includes one of the processes most commonly used by industries today: bioremediation. Bioremediation makes possible the recovery of oil contaminated sites through biostimulation, which increases the hydrocarbon-degrading ability of native microorganisms, thereby transforming or mineralizing the pollutants.

Quantification of Salmonella in asymptomatic carrier animals can be used to assess microbial risk and monitor the level of contamination in domestic animals used for food production. We examined the sensitivity, specificity and accuracy of real-time qPCR, without pre-enrichment or selective enrichment stages, for the quantification of S. enterica serovar Enteritidis in resistant mice, as a model of asymptomatic carrier animal. The results were compared with those obtained by traditional bacteriological culture methods, the gold standard test. Two hundred and forty-three samples, including spleen, liver, mesenteric lymph nodes, portions of intestine, intestinal content of the ileocecal portion, and feces, were collected from a group of 27 C57BL/6 mice, that had been intragastrically inoculated with high doses of S. enterica serovar Enteritidis. The real-time qPCR assay presented a consistent linearity of the standard curve (r(2) = 0.999), with very low differences between melting temperatures, and low coefficients of variation in intra- (< 1%) and interassay (< 2%) comparisons. The primers were highly specific; there was no amplification with other Salmonella serovars or with DNA from uninfected tissues and feces from mice. The detection limit of the technique was defined as 32 copies of S. enterica serovar Enteritidis. A sensitivity of 90%, a specificity of 77% and an accuracy of 79% were obtained. The higher sensitivity of PCR was reflected in a kappa coefficient of 0.41, showing moderate agreement between tests. We conclude that real-time qPCR is a good alternative for diagnostic scanning in asymptomatic carrier animals, due to its high sensitivity and rapidity.

The goal of this study was to assess the presence and surfactant potential of naturally occurring microbes from a tropical soil with petrochemical contamination. Microorganisms in a soil sample from a Brazilian landfarm were isolated and grown on petroleum as the sole carbon source. Of 60 isolates screened for petroleum-based growth, 7 demonstrated surfactant activities by the drop–collapse methodology over various types of oils. From their growth profiles in liquid culture during 132 h, all had their first detection of surfactant activity after 96 h. Little is currently known about biosurfactant-producing microorganisms in tropical environments contaminated by hydrophobic compounds, and the search for them is essential for bioremediation and for oil recovery enhanced by microbes. Our results indicate that different petroleum-grown microorganisms showing surfactant activity can be recovered from landfarm soil in a tropical environment.

Amostras fecais de cães provenientes de distritos carentes do Município de Ilhéus / BA foram analisadas para a presença de Salmonella spp durante o período de junho de 2001 a abril de 2002. Dezoito (9,47%) dos 190 animais foram considerados positivos sendo que 66,6% destes casos ocorreram com animais até 1 ano de idade, 56% se alimentavam com comida caseira, 66% tinham acesso à fonte de água não tratada e 67% possuía comportamento domiciliar. A sintomatologia da doença não foi percebida em 83,3% dos animais. A S. Gafsa representou o sorotipo mais prevalente (38,9%), seguido pela S. Rubislaw (27,8), S. Carrau (16,7%) e S. enterica subsp. Houtenae (11,1%). Uma cepa não pôde ser identificada. O antibiograma revelou a gentamicina como sendo a droga mais potente, in vitro, para o tratamento das salmonelas encontradas na região de Ilhéus, seguida pela ampicilina. Nossos resultados confirmam que cães representam um importante reservatório de sorotipos exóticos de Salmonella e que ocorre padrões diferentes de sensibilidade a antibióticos entre eles.