Direct Detection and Quantification of Bacterial Pathogens from Broiler Cecal Samples in the Slaughter Line by Real-Time PCR

Chicken meat is an important source of foodborne pathogens, including Salmonella, Campylobacter, and Clostridium perfringens. These bacteria can occur in the intestinal microbiota of broilers and contaminate chicken carcasses in industrial meat processing. This study aimed to develop and evaluate a procedure based on real-time PCRs for the direct detection and quantification of these three bacteria in broilers' ceca collected in poultry slaughter houses and demonstrate the occurrence of these important foodborne pathogens in Brazilian poultry production flocks. Cecal contents were collected from 45 different broiler flocks in three different slaughterhouses in the state of Paraná, Brazil, totaling 45 samples (in pools of 10 different ceca/chickens per broiler flock). Then, these samples were tested for the detection and quantification of Salmonella, Campylobacter, and Clostridium perfringens by real-time PCRs. The results demonstrated the occurrence of three (6.7%) positive pools for Salmonella, 20 (44.4%) for Campylobacter, and 32 (71.1%) for C. perfringens. Mean bacterial concentrations in the positive samples were 4.3log10 cells/g for Salmonella, 6.4 log10 cells/g for Campylobacter, and 5.5 log10 cells/g for C. perfringens. In conclusion, Salmonella, Campylobacter, and C. perfringens could be detected and quantified directly from the broilers cecal contents collected in the slaughter line. This procedure will be certainly useful to more quickly detect these foodborne pathogens and prevent their occurrence in chicken meat and other poultry food products.(AU)

Photohydroionisation for disinfection of poultry litter.

1. This study evaluated photohydroionisation efficiency on the disinfection of new shavings used as substrate for litter in the poultry industry, pre-inoculated with bacterial, fungal and viral agents.2. Each replicate consisted of 250 g of new shavings sterilised by autoclaving, challenged with bacterial (Escherichia coli, Staphylococcus aureus, and Salmonella enterica, serovar Abony), fungal (Saccharomyces cerevisiae) and viral inoculum (Gumboro disease virus). The experiment consisted of six replicates at four times (0, 1, 5 or 10 min exposure) of photohydroionisation. The disinfection process was performed in a bench photohydroionisation device with four ultraviolet lamps. The agents inoculated in the shavings were analysed after the disinfection process.3. The counts of enterobacteria and total bacteria showed a quadratic effect. In contrast, the counts of fungi and viruses showed a negative linear effect with an increase in the time of photohydroionisation. The enterobacteria showed a linear response plateau effect (LRP), with a minimum time point of 5.498 minutes at a minimum contamination of 0.666 CFU/g and a reduction of 82.27% of the pre-established inoculum. Total bacteria had an LRP effect with a minimum time point of 1.902 minutes at minimum contamination of 1.739 CFU/g and a reduction of 50.0% of the pre-established inoculum. An LRP effect was found for fungi, with a minimum time point of 7.931 minutes in minimum contamination of 3.380 CFU/g, and with a reduction of 11.0% of the pre-established inoculum. For viruses, there was an LRP effect with a minimum time point of 5.012 minutes in minimum contamination of 0.000 viral titre per 100 g of shavings, which was reduced by 100% of the pre-established inoculum.4. Photohydroionisation in the disinfection of new shavings used as poultry litter has partial potential as a microbiological control tool, as a complete reduction occurred only for the viruses, whereas for bacteria and fungi only partial reductions of these microorganisms were observed.

Lack of association between human leukocyte antigen polymorphisms rs9277535 and rs7453920 and chronic hepatitis B in a Brazilian population.

Hepatitis B virus (HBV) infection is a serious public health problem worldwide. The progression of the disease depends on several host and viral factors and may result in fulminant hepatitis (very rare), acute hepatitis with spontaneous clearance, and chronic hepatitis B infection. Previous studies demonstrated that variations in the human leukocyte antigen (HLA) class II (HLA-DPB1 and HLA-DQB2 genes) are related to the chronic HBV infection. This study aimed to investigate the association of two single nucleotide polymorphism (SNPs), one in the HLA-DPB1 (rs9277535) and one in the HLA-DQB2 (rs7453920), with chronic hepatitis B infection in a southern Brazilian sample. This case-control study included 260 HBV patients attended in a Specialized Center for Health in Caxias do Sul (Brazil) between 2014 and 2016. The same number of controls (matching for age, gender, and ethnicity) was obtained in a University Hospital in the same city and period. Blood samples were collected and genomic DNA was extracted. Genotyping were performed by real-time Taqman PCR method. Odds ratios with 95% confidence intervals and significance level of 5% (P < 0.05) were calculated. Allele frequencies in the SNP rs9277535 were 72.6% for A and 27.4% for G nucleotides in cases and 75.0% for A and 25.0% for G in controls. Allele frequencies in the SNP rs7453920 were of 25.7% for A and 74.3% for G in cases and 28.8% for A and 71.2% for G in controls. No statistically significant association was found between both SNPs and chronic hepatitis B (P > 0.05).

Improving household air, drinking water and hygiene in rural Peru: a community-randomized-controlled trial of an integrated environmental home-based intervention package to improve child health.

Background: Diarrhoea and acute lower respiratory infections are leading causes of childhood morbidity and mortality, which can be prevented by simple low-cost interventions. Integrated strategies can provide additional benefits by addressing multiple health burdens simultaneously. Methods: We conducted a community-randomized-controlled trial in 51 rural communities in Peru to evaluate whether an environmental home-based intervention package, consisting of improved solid-fuel stoves, kitchen sinks, solar disinfection of drinking water and hygiene promotion, reduces lower respiratory infections, diarrhoeal disease and improves growth in children younger than 36 months. The attention control group received an early child stimulation programme. Results: We recorded 24 647 child-days of observation from 250 households in the intervention and 253 in the attention control group during 12-month follow-up. Mean diarrhoea incidence was 2.8 episodes per child-year in the intervention compared with 3.1 episodes in the control arm. This corresponds to a relative rate of 0.78 [95% confidence interval (CI): 0.58-1.05] for diarrhoea incidence and an odds ratio of 0.71 (95% CI: 0.47-1.06) for diarrhoea prevalence. No effects on acute lower respiratory infections or children's growth rates were observed. Conclusions: Combined home-based environmental interventions slightly reduced childhood diarrhoea, but the confidence interval included unity. Effects on growth and respiratory outcomes were not observed, despite high user compliance of the interventions. The absent effect on respiratory health might be due to insufficient household air quality improvements of the improved stoves and additional time needed to achieve attitudinal and behaviour change when providing composite interventions.

Reproductive biology of the epiphytic bromeliad Werauhia gladioliflora in a premontane tropical forest.

The floral phenology, fruit and seed production, and self-compatibility of Werauhia gladioliflora, an epiphytic bromeliad with a wide distribution, were studied in a premontane forest in the Monteverde area in Costa Rica. The species presents the pollination syndrome of chiropterophily, and it is visited by the small bats Hylonycteris underwoodi and Glossophaga commissarisi (Glossophaginae). The population flowering period extended from October to early December (end of rainy season) and seed dispersal occurred from February to April (dry season). Most plants opened a single flower per night, either every day or at one-day intervals during the flowering period. In natural conditions, the average fruit set amounted to almost half of the potential output, but individual fecundity (number of seeds) remained high. Seed number per fruit and germination capacity after artificial selfing and out-crossing treatments did not differ from natural pollination conditions. Werauhia gladioliflora exhibited high levels of autonomous self-pollination and self-compatibility at the individual and population level, characters associated with the epiphytic habitat. These reproductive traits are also associated with early colonizer species, yet life history traits, such as seed dispersal, seedling establishment success, and growth, are likely to have a major role in determining the presence of this species in the successional vegetation patches scattered over the studied premontane area.

Synthesis of catecholamide spiroarsoranes and their in vitro anthelmintic properties against Molinema dessetae and Nippostrongylus brasiliensis infective larvae.

Catecholamide spiroarsoranes were synthesized and evaluated for anthelmintic properties on two in vitro models, infective larvae of the filaria Molinema dessetae and infective larvae of an intestinal nematode. Nippostrongylus brasiliensis. On the N dessetae model, the most active compound after 24 h incubation time had an EC50 of 0.1 mumol/l. Eleven compounds had EC50 's in a range from 2 to 200 mumol/l. After 7 days incubation time, the two most active compounds had EC50 's of 0.03 and 0.07 mumol/l, respectively. On the N brasiliensis model, only three compounds were slightly active after 4 days incubation time. The ligands used for the spiroarsoranes synthesis were also evaluated for anthelmintic activity in order to know the contribution of these structures in the spiroarsoranes activity. Spiroarsoranes as prodrugs of arsonic acids were very active on the filaria, nematode having predominantly transcuticular uptake of nutrients while the activity on the intestinal nematode having both the types, transcuticular and intestinal uptake was low. The high sensitivity of filarial infective larvae is probably in relation to their location in the mosquito whereas N. brasiliensis infective larvae are telluric and should be more unsensitive to survive in a variable environment.

Selective damage to either the "M" or "P" cell pathway in human retinal disease: implications for visual processing.

Studies in primates have shown there to be 2 main visual afferent pathways (magno and parvo) operating in parallel with different functional roles. Disease state may selectively affect one or other of these, as shown by particular sensory losses. In cases of treated Cuban tropical "neuropathy" (a retinopathy also occurs) visual acuity and achromatic contrast sensitivity are preserved although colour vision is severely affected indicating selective preservation of the "magno" pathway. In Melanoma Associated Retinopathy, red-green colour vision is preserved although achromatic contrast sensitivity for low spatial frequencies, and the detection of motion is damaged suggesting preferential damage to the "magno" pathway. Our studies illuminate two controversial aspects of the P and M systems: the role of type II parvocellular cells in colour vision and the role of M cells in detecting stimuli of low spatial and low temporal frequency.

Streptococcal toxic shock-like syndrome.

BACKGROUND: After decades marked by a decrease in the incidence and severity of scarlet fever and streptococcal soft-tissue infections, invasive infections with group A streptococci have reemerged as a global public health problem. Sporadic outbreaks of a rapidly progressive disorder characterized by fever, shock, desquamating rash, and multiorgan system failure often associated with severe suppurative soft-tissue infection have recently been recognized in young, otherwise-healthy adults. Referred to as streptococcal toxic shock-like syndrome, this acute, progressive, often fatal illness appears to be related to an overall change in the virulence of group A streptococci and the reappearance of highly mucoid exotoxin-producing strains. However, the relationship between virulence factors, epidemiological features, and precise pathogenesis of these infections remains unclear. OBSERVATIONS: The 80% incidence of soft-tissue involvement in streptococcal toxic shock-like syndrome and the high frequency of progression to necrotizing fasciitis, myositis, or even death are unprecedented. The type and extent of tissue injury, systemic toxicity, and multiorgan system failure associated with this syndrome appear to be mediated by streptococcal pyrogenic exotoxins via the induction of biologically potent endogenous cytokines. The observed increase in the incidence and severity of invasive group A streptococcal infections coincides with the resurgence of genetically related, highly virulent strains of Staphylococcus pyogenes that express an invasive phenotype and carry a specific gene for pyrogenic exotoxin A. CONCLUSIONS: The changing spectrum of invasive group A streptococcal infections has prompted a reexamination of the microorganism and its diverse clinical manifestations. A clonal basis for increased virulence expression is supported epidemiologically. As the incidence of streptococcal toxic shock-like syndrome appears to be increasing, it is imperative that clinicians become familiar with the presentation of this syndrome so they can diagnose and treat it in a timely and effective manner.

Human milk secretory IgA antibody to nontypeable Haemophilus influenzae: possible protective effects against nasopharyngeal colonization.

Sixty-eight children fed human milk were followed prospectively from birth to 12 months of age to assess the effect of milk antibody on nasopharyngeal colonization. Human milk secretory IgA antibody to P6, a highly conserved outer membrane protein of nontypeable Haemophilus influenzae, was measured with the use of an enzyme-linked immunosorbent assay. Nasopharyngeal colonization with nontypeable H. influenzae and the occurrence of otitis media were determined. Nasopharyngeal colonization was found in 22 children (32%), and 39 children (57%) had otitis media. Frequency of isolation of nontypeable H. influenzae was directly related to episodes of otitis media (r = 0.35; p = 0.001). The level of human milk anti-P6 secretory IgA antibody was inversely related to frequency of isolation of the organism (r = -0.27; p = 0.026). The average antibody level, expressed as nanograms per 0.1 mg total secretory IgA, in human milk fed to children with no colonization of nontypeable H. influenzae was significantly higher than in milk fed to children in whom colonization occurred on multiple occasions (156 +/- 120 vs 69 +/- 50; p = 0.013). Prevention of colonization was most evident during breast-feeding. These data suggest that the protective effects of human milk against otitis media may be due in part to inhibition of nasopharyngeal colonization with nontypeable H. influenzae by specific secretory IgA antibody.