Recreational sandboxes for children and dogs can be a source of epidemic ribotypes of Clostridium difficile

Different studies have suggested that the sand of public playgrounds could have a role in the transmission of infections, particularly in children. Furthermore, free access of pets and other animals to the playgrounds might increase such a risk. We studied the presence of Clostridium difficile in 20 pairs of sandboxes for children and dogs located in different playgrounds within the Madrid region (Spain). Clostridium difficile isolation was performed by enrichment and selective culture procedures. The genetic (ribotype and amplified fragment length polymorphism [AFLP]) diversity and antibiotic susceptibility of isolates was also studied. Overall, 52.5% (21/40) of samples were positive for the presence of C. difficile. Eight of the 20 available isolates belonged to the toxigenic ribotypes 014 (n = 5) and 106 (n = 2), both regarded as epidemic, and CD047 (n = 1). The other 12 isolates were non‐toxigenic, and belonged to ribotypes 009 (n = 5), 039 (n = 4), and 067, 151 and CD048 (one isolate each). Nevertheless, all isolates (even those of a same ribotype) were classified into different AFLP genotypes indicating non‐relatedness. In conclusion, our results revealed the presence of epidemic ribotypes of C. difficile in children's and dog's sandboxes located nearby, which constitutes a major health risk.     

Plasma ferritin of sows during pregnancy and lactation

We studied a set of indicators of iron metabolism in sows and new-born piglets: hemoglobin, plasma iron, total iron binding capacity and plasma ferritin. Iron status of seventy-two sows was studied throughout pregnancy and lactation. The results point to mobilization of iron from its storage sites up to the second month of gestation, but no appreciable decrease during the second half of pregnancy. No appreciable mobilization of the iron stores of sows was observed during the lactation period. Nine sows and their 78 piglets were used to compare the status of iron in the sows and the end of gestation with the iron status of the piglets at birth; there was a positive correlation between the mean values of plasma ferritin concentration in the piglets and the levels of plasma ferritin in their mothers.     

Molecular characterization of reticuloendotheliosis virus insertions in the genome of field and vaccine strains of fowl poxvirus

Evidence of the widespread occurrence of reticuloendotheliosis virus (REV) sequence insertions in fowl poxvirus (FPV) genome of field isolates and vaccine strains has increased in recent years. However, only those strains carrying a near intact REV provirus are more likely to cause problems in the field. Detection of the intact provirus or REV protein expression from FPV stocks has proven to be technically difficult. The objective of the present study was to evaluate current and newly developed REV and FPV polymerase chain reaction (PCR) assays to detect the presence of REV provirus in FPV samples. The second objective was to characterize REV insertions among recent "variant" FPV field isolates and vaccine strains. With REV, FPV, and heterologous REV-FPV primers, five FPV field isolates and four commercial vaccines were analyzed by PCR and nucleotide sequence analysis. Intact and truncated REV 5' long terminal repeat (LTR) sequences were detected in all FPV field isolates and vaccine strains, indicating heterogeneous REV genome populations. However only truncated 3' LTR and envelope sequences were detected among field isolates and in one vaccine strain. Amplifications of the REV envelope and 3' LTR provided strong evidence to indicate that these isolates carry a near intact REV genome. Three of the four FPV vaccine strains analyzed carried a solo complete or truncated 5' LTR sequence, indicating that intact REV provirus was not present. Comparison of PCR assays indicated that assays amplifying REV envelope and REV 3' LTR sequences provided a more accurate assessment of REV provirus than PCR assays that amplify the REV 5' LTR region. Therefore, to differentiate FPV strains that carry intact REV provirus from those that carry solo 5' LTR sequences, positive PCR results with primers that amplify the 5' LTR should be confirmed with more specific PCR assays, such as the envelope, or the REV 3' LTR PCR.     

Temperature and storage period over spermatic parameters of jundiá, Rhamdia quelen (Quoy & Gaimard, 1824)

The aim of this work was to evaluate the spermatic parameters of jundia, Rhamdia quelen, semen stored for short periods under different temperatures. Fifteen males were used in a time factorial experimental design (13 × 3 × 3 × 3). The sperm was stored at: 15; 25 and 35°C and activated in water at: 15; 25 and 35°C each, respectively, within storage periods of: 0; 2; 4; 6; 8; 10; 12; 16; 20; 24; 32; 40 and 48 h after collection. The treatments were performed in triplicates and in sequential protocols every 50 h. The motility parameters were evaluated using Pearson's correlation analysis and the significant parameters (P < 0.05) were summarized using principal component analysis forming two predicted groups. Group 01 composed the following parameters: curvilinear velocity, wobble and linearity; group 02 comprised average path velocity, straight line velocity, beating cross frequency and motility rate. Effects (P < 0.05) of treatments were observed only in group 02, with interaction between time and temperature of exposure and between exposure time and water temperature. Superior results of spermatic parameters of group 02 were observed right after collection. However, the exposure and spermatic activation at 15°C assured significant results 48 h after collection when compared with 25 and 35°C.     

Effect of biofloc technology (BFT) on the early postlarval stage of pink shrimp <Emphasis Type="Italic">Farfantepenaeus paulensis</Emphasis>: growth performance,floc composition and salinity stress tolerance

Biofloc rearing media provides a potential food source for shrimp reared in limited or zero water exchange systems. This culture system is environmentally friendly as it is based on limited water use and minimal effluent is released into the surrounding environment. In this study, we evaluated the survival, growth performance and salinity stress tolerance of pink shrimp Farfantepenaeus paulensis postlarvae reared from PL₁₀ to PL₂₅ in a biofloc technology limited water exchange system. PL (mean ± SD weight and length of 14 ± 10 mg and 8.10 ± 0.7 mm, respectively) were reared in nine 40-L plastic tanks with a stocking density of 10PL/L. Three culture treatments were applied (1) culture in the presence of bioflocs and commercial feed supply (FLOC + CF); (2) culture in the presence of biofloc without feed supply (FLOC) and (3) culture in clear water with feed supply (control). Final biomass and survival were significantly higher in FLOC + CF treatment than the control (P < 0.05), but did not differ from FLOC. PL reared in the FLOC + CF treatment achieved a significantly higher final weight, weight gain and length in comparison with the other two treatments (P < 0.05). No significant difference (P > 0.05) between treatments was found for salinity tolerance over 24 and 48 h durations. The proximate analysis of floc shown high levels of crude protein (30.4%), but low levels of crude lipids (0.5%). The continuous availability of bioflocs had a significant effect on growth and survival of F. paulensis postlarvae cultured in BFT nursery systems.     

Mineral contents of the muscle tissue and the digestive gland of Octopus vulgaris during short‐term starvation

We used spectrometry to study variation in the contents of minor (B, Cu, Fe, Mn and Zn) and major essential elements (Ca, K, Mg, Na and P) in the muscle tissue and digestive glands of Octopus vulgaris after 1, 2, 4 or 8 days of starvation and in fed controls. The contents of the major essential elements remained constant during 8 days of starvation. Na was the principal element found in both tissues, followed by K, P and Mg. The contents of the minor essential elements were higher in the digestive gland than in the muscle tissue, with increased contents of Fe, Mn and Zn (p < .05) during starvation, confirming the high capacity of this organ to accumulate trace minerals. In muscle tissues, B, Fe and Mn contents were lower and Cu and Zn contents higher, in starved animals than in fed animals (p < .05). Significant negative correlations between the digestive gland index and the contents of Cu, Fe and Zn suggested that the loss of weight of the digestive gland during starvation is responsible for this increase in mineral concentration. This and further information on variation in mineral content will help to optimize performance of diets formulated for the octopus.     

Identification and characterization of lactic acid bacteria isolated from rainbow trout, Oncorhynchus mykiss (Walbaum), with inhibitory activity against Lactococcus garvieae

A study was conducted to evaluate the probiotic properties of endogenous rainbow trout microbiota against pathogenic Lactococcus garvieae. A total of 335 bacterial strains were isolated from rainbow trout and screened for antagonistic activity against L. garvieae using an agar spot assay. Antagonistic strains were grouped by PCR amplification of repetitive bacterial DNA elements (rep‐PCR) and identified by 16S rRNA gene sequence analysis. The results revealed that the antagonistic strains belonged to the genera Lactobacillus, Lactococcus and Leuconostoc. Further probiotic characteristics, such as specific growth rate, doubling time, resistance to biological barriers, antibiotic resistance, hydrophobicity and production of antimicrobial substances, were also studied. These strains were able to survive low pH and high bile concentrations, showed good adherence characteristics and a broad spectrum of antibiotic resistance. The antagonistic efficacy was maintained after sterile filtration and was sensitive to proteinase K, indicating that proteinaceous extracellular inhibitory compounds were at least partially responsible for pathogen antagonism. Based on these results, these strains should be further studied to explore their probiotic effects in challenge experiments in vivo. This study shows clear evidence that the indigenous trout‐associated microbiota may provide a defensive barrier against L. garvieae.     

Intensive rearing of juvenile crayfish (Pacifastacus leniusculus,Astacidae) during the first 6 months: effects of size grading

Recent advances in crayfish feeding have enabled the development of size grading studies from the start of first‐feeding. A 180‐day experiment aimed at intensive rearing of Pacifastacus leniusculus was carried out under controlled conditions, evaluating the effects of size grading at two different periods from the onset of exogenous feeding. Stage 2 juveniles were stocked in fibreglass tanks at a density of 100 m², and fed a dry diet for salmonids combined with restricted amounts of Artemia cysts. Five groups were tested: no grading, grading at 60 days (large and small size) and grading at 100 days (large and small size). After 6 months, no significant differences were found in the survival among groups (mean: 73.06%). The highest final growth (pooled results from upper and lower classes: 17.39 mm carapace length, 1.43 g weight) was achieved by the crayfish sorted at 60 days, showing significant differences from the ungraded group. Smaller crayfish graded at 60 days grew significantly faster than smaller crayfish graded at 100 days. The food conversion ratio was lower in the graded groups (mean: 2.64), showing significant differences from the ungraded group (3.23). This study shows that size grading allows a better performance and an improved feeding efficiency.     

Turbine entrainment and passage of potadromous fish through hydropower dams: Developing conceptual frameworks and metrics for moving beyond turbine passage mortality

Potadromous fishes are vulnerable to involuntary entrainment through hydropower turbines. However, turbines can also provide a downstream passage route for potadromous fish. Here, we review evidence for turbine entrainment and passage in potadromous fish, and evaluate the effects of these processes on upstream and downstream populations. We develop conceptual frameworks and metrics to quantify vulnerability to turbine entrainment removals, and to quantify the efficiency of turbines as a downstream passage route. We highlight factors that influence these processes and provide case‐studies demonstrating their applicability. We found that juvenile potadromous fish are being entrained through turbines at rates high enough to impact upstream populations. Given that juvenile passage survival is often high, we argue that turbines provide an important downstream passage route for potadromous fish. We show that entrainment vulnerability is likely a function of interactions between in‐reservoir fish behaviour, habitat configuration and operations and thus not well captured by passage mortality estimates. Similarly, we show that while passage mortality can limit downstream passage efficiency, passage success is also dependent on reservoir and forebay navigation, along with survival and fitness in the downstream river. We advocate for a shift in focus away from estimates of passage mortality and injury, which have previously accounted for the majority of turbine passage research. Instead, we recommend an approach that focusses on quantification of the factors that influence downstream passage efficiency and entrainment vulnerability. Moreover, we highlight the need to better understand the broader scale impacts of these events on upstream and downstream populations.     

Detection of hepatitis E virus shedding in feces of pigs at different stages of production using reverse transcription-polymerase chain reaction.

The aim of this study was to determine at which production stages hepatitis E virus (HEV) is shed by the highest number of pigs and to estimate the relative risk associated with each stage. For this purpose, 146 fecal samples of pigs from 21 farms were studied. In addition, 1 sample from the manure ditch and another sample of drinking water, collected directly from the trough located in the pen, were taken from 16 farms. HEV RNA was detected in fecal samples from 34 pigs (23.29%). The production stages in which most pigs excreted HEV were weaners (41.7%) and pigs in the first month of feeding (60%). The results of the statistical analysis showed that the principal significant risk stage in HEV shedding was the first month of feeding (odds ratio [OR] 19.5, 95% CI 3.59-106.07, P = 0.001) followed by the weaners stage (OR 9.3, 95% CI .78-48.42, P = 0.008). In 8 out of 16 farms tested (50%) HEV RNA was detected in raw manure and in the water trough of only 1. Detection of HEV in manure ditches raises the concern of how to deal with manure of swine origin, because it is used as soil fertilizer.