Influence of porosity of low-density polyethylene media on the maturation process of biofilters used in recirculating aquaculture systems

The effect of pores with different diameters in polyethylene media on the process of ammonia nitrification during the maturation of biofilters is evaluated. Four types of media were produced: M1 without pores, M2 with 40–112 μm in diameter pores, M3 with 125–187 μm diameter pores, and M4 with 312–437 μm diameter pores. Ammonia concentrations were statistically equal among media. On the majority of days, nitrite concentrations were lowest for M1 and M2, intermediate for M3, and highest for M4. Increased pore diameter favored the formation of biofilm in the interior, but the pores obtained in the expansion process were not interconnected, which may have caused low oxygenation of the water in the internal area. It was concluded that porosity did not reduce biofiltration time and did not increase the efficiency of nitrification of ammonia.     

Weathering,Biomass Production and Groundwater Chemistry in an Area of Dominant Anthropogenic Influence,the Chaves-Vila Pouca de Aguiar Region,North of Portugal

Using a combination of a grouping algorithm and a geochemical mole balance algorithm, we could identify and quantify the contributions made by water-rock and water-biomass interactions to the composition of shallow groundwaters in northern Portugal, in an area with a sizable anthropogenic input (the Chaves-Vila Pouca de Aguiar region). The first algorithm, based on the mathematical concept of equivalent relations, allows for the grouping of water samples with similar chemistries. The second algorithm, which uses the stoichiometries and mole/charge balances in weathering reactions and biological processes, provides the possibility to relate the average chemistry of each identified group to water-mineral and water-biomass interactions. This algorithm hinges on ratios of dissolved silica and bicarbonate, constituents considered to be derived only from chemical weathering and biomass production. Background information on the petrology, mineralogy and chemistry of bedrocks and soils, on climatic differences, on the history of deforestation and reforestation of the study area, and on the application and composition of fertilizers and manure, was used to identify the most realistic weathering reactions and biological processes which contribute to the water chemistry. The concentrations of chloride, sulphate and nitrate were considered to represent sources other than chemical weathering or biomass changes, like cyclic salts, fertilizers and manure. Despite the high background concentrations derived from such sources, we were able to quantify the contributions of the identified weathering reactions and botanical uptake to the groundwater composition. The spatial distributions of the various groundwater groups shows a good correlation with the geology, land use and climatic differences in the area.     

Heterotic groups based on yield-specific combining ability data and phylogenetic relationship determined by RAPD markers for 28 tropical maize open pollinated varieties

Twenty eight maize open pollinated varieties (OPVs) were crossed in a diallel scheme and the 378 F₁'s were evaluated in 10 environments in Brazil. Based on yield-specific combining ability data (SCA), these varieties were classified in four heterotic groups. The consistence of the proposed heterotic groups was confirmed comparing intra- and inter-group F₁ values and midparent heterosis. Superior OPVs combinations for use as a source of inbreds in hybrid breeding programs were determined. RAPD markers were used to genotype these varieties. A UPGMA dendogram, based on marker data from 50 primers and 178 polymorphic bands, was obtained. Phylogeny obtained with RAPD markers agreed with known pedigree data. Dent germplasm tended to group separately from flint germplasm. Multidimensional Scaling Analysis on marker data and morphological data showed a higher degree of genetic divergence among the dent germplasm than among the flint germplasm used in this study. Correlation between RAPD marker estimated genetic distance and SCA for yield was low and positive (r = 0.16^**). This revised version was published online in August 2006 with corrections to the Cover Date.     

Selenium and aflatoxin levels in raw Brazil nuts from the Amazon basin

Whereas selenium (Se) is an important antioxidant in human metabolism to prevent cancer, aflatoxins are highly carcinogenic. Brazil nuts from Eastern and Western Amazon regions were evaluated to find any relationship between Se and aflatoxins levels. A total of 80 (in-shell and shelled) nuts samples were collected directly from different forest sites and analyzed for Se by atomic emission spectrometry and aflatoxins by liquid chromatography tandem mass spectrometry. The limit of quantitation (LOQ) for Se was 2.0 mg/kg, and LOQ for total aflatoxins was 0.390 microg/kg. Nut Se levels from the Eastern region were higher than the Western, in addition to the aflatoxins. The moisture content (mc) and water activity (aw) of the raw nuts from the two regions did not present a significant difference, for either in-shell or shelled. The mc was 24.5% (minimum of 20.1% and maximum of 30.4%) and 22.1% (minimum of 14.6% and maximum of 28.9%) and a w of 0.85 for both regions. Further studies need to be carried out to discover the role of Se on fungi growth stress and aflatoxin production mechanisms.     

Development of a tandem repeat-based multilocus typing system distinguishing Babesia bovis geographic isolates

Mini- and microsatellite sequences have proven to be excellent tools for the differentiation of strains and populations in several protozoan parasites due to their high variability. In the present work we have searched the genome of the tick-transmitted bovine hemoprotozoon Babesia bovis for tandem repeats (TRs) that could be useful for a multilocus typing system. Hundred and nineteen sequences were shortlisted and tested in five common B. bovis reference isolates originating from distinct geographic locations of North and South America: Texas, USA (T2Bo), Mexico (RAD and Mo7), and Santa Fe and Salta, Argentina (R1A and S2P, respectively). Satellite sequences were PCR-amplified using specific primers, separated by polyacrylamide gel electrophoresis, visualized by silver staining and sized. Fourteen TR sequences could be reliably amplified in all isolates and displayed length polymorphism. All primers used were specific for B. bovis and did not amplify genomic DNA from the bovine host or from Babesia bigemina, the principal co-infecting bovine parasite in the Americas, allowing their future use in field surveys. The 14 satellite markers identified are distributed throughout the four chromosomes of B. bovis as follows: chromosome 1 (n = 3), chromosome 2 (n = 2), chromosome 3 (n = 5), and chromosome 4 (n = 4). Within the five B. bovis isolates we identified nine satellite marker loci with two alleles, three with three alleles, one with four and another with five alleles. In comparison to Theileria parva, a bovine hemoprotozoan that pertains to the same piroplasmida order and owns a genome of similar size, the number of polymorphic TRs and the average number of alleles per TR locus seem to be significantly reduced in the B. bovis genome. Furthermore, the ratio of micro- to minisatellites in both B. bovis and T. parva is considerably lower than in other eukaryotes, as confirmed by bioinformatic analysis. The multilocus genotype of the five B. bovis isolates was assessed and the genetic distance between each other determined followed by cluster analysis based on neighbor joining. The resulting phenogram showed that B. bovis isolates segregated into three clusters according to their geographic origin. The presented marker system is suitable to explore various parameters of B. bovis populations such as genetic diversity, infection dynamics and their structure under different epidemiological situations, which are of crucial importance for improved control strategies.     

Concurrent avian malaria and avipox virus infection in translocated South Island saddlebacks (Philesturnus carunculatus carunculatus)

Abstract

CASE HISTORY: Outbreaks of mortality in South Island saddlebacks (Philesturnus carunculatus carunculatus) that had been translocated to two offshore islands in the Marlborough Sounds of New Zealand were investigated during the summer of 2002 and 2007. Both outbreaks were associated with a severe decrease in numbers of saddlebacks of up to 60% of approximately 200 birds.

CLINICAL AND PATHOLOGICAL FINDINGS: Many of the surviving birds were in poor condition, and had skin lesions on the legs and head. Necropsy showed pale liver and lungs, and a swollen spleen. Histopathology revealed schizonts resembling Plasmodium spp. within the cytoplasm of many hepatocytes and splenic histiocytes. The skin lesions consisted of epithelial proliferations containing numerous Bollinger bodies typical of avipox virus (APV) infection. Two different APV were isolated, using PCR, from two different birds exhibiting skin lesions. Each isolate had 100% sequence homology with APV members from either Clade A or Clade B. In addition, PCR analysis revealed that the Plasmodium elongatum present in infected birdsbelonged to a strain that was endemic in the population of North Island saddlebacks (Philesturnus carunculatus rufusater).

DIAGNOSIS: Concurrent infections with Plasmodium spp. haemoparasites and APV were identified as the likely cause of death in the birds examined.

CONCLUSIONS AND CLINICAL RELEVANCE: Although the Plasmodium spp. identified is thought to be endemic to saddlebacks in New Zealand, the affected birds were likely to be immunocompromised by concurrent APV infection or through lack of genetic diversity. Both the introduced mosquito Culex quinquefasicatus and the native mosquito Culex pervigilans are likely vectors for both these diseases, and the provision of water supplies less favourable to mosquito-breeding is recommended.     

Inactivation of Ichthyophonus spores using sodium hypochlorite and polyvinyl pyrrolidone iodine

Chlorine and iodine solutions were effective at inactivating Ichthyophonus spores in vitro. Inactivation in sea water increased directly with halogen concentration and exposure duration, with significant differences (P < 0.05) from controls occurring at all chlorine concentrations and exposure durations tested (1.5–13.3 ppm for 1–60 min) and at most iodine concentrations and exposure durations tested (1.2 ppm for 60 min and 5.9–10.7 ppm for 1–60 min). However, 10‐fold reductions in spore viability occurred only after exposure to halogen solutions at higher concentrations and/or longer durations (13 ppm total chlorine for 1–60 min, 5.9 ppm total iodine for 60 min, and 10.7 ppm total iodine for 1–60 min). Inactivation efficacy was greater when halogen solutions were prepared in fresh water, presumably because of combined effects of halogen‐induced inactivation and general spore instability in fresh water. The results have practical implications for disinfection and biocontainment in research laboratories and other facilities that handle live Ichthyophonus cultures and/or infected fish.     

The Effects of Cryopreservation on the Morphometric Dimensions of Iberian Red Deer (Cervus elaphus hispanicus) Epididymal Sperm Heads

Computer-automated sperm-head morphometry was used in this study to determine the effects of cryopreservation on red deer sperm-head morphometry. Epididymal sperm samples were collected from 40 mature stags and were divided. One portion was diluted at room temperature in a Tris-citrate egg yolk medium, containing 6% glycerol. A microscope slide was prepared from single extended sperm samples prior to freezing. The remainder of each sample was frozen in nitrogen vapours. After thawing, sperm smears were prepared as described above. All slides were air dried and stained with Hemacolor. The sperm-head dimensions for length, width, area, perimeter and shape factor (length/width), for a minimum of 135 spermatozoa were determined for each slide by means of the Sperm-Class Analyser (SCA). Firstly, our results show that cryopreservation substantially reduced (p < 0.001) sperm motility and plasma membrane and acrosome integrities. In addition, sperm heads were significantly smaller in cryopreserved spermatozoa than in the companion extended samples for area (32.05 microm2 vs 32.56 microm2; p < 0.05), length (8.46 microm vs 8.53 microm; p < 0.0001) and shape factor (1.833 vs 1.849; p < 0.0001) for all stags. These differences were found within 29 of 40 stags (75%) for at least three of the morphometric parameters. The individual variability (CV) of sperm head measurements from extended samples was negatively correlated (p < 0.005) with the per cent of change in sperm head measurements after cryopreservation for area (r = -0.465), width (r = -0.483) and perimeter (r = -0.375). Thus, the lower the sperm head variability in the extended samples, the greater the sperm change as a consequence of the cryopreservation. These results suggest that the variability (heterogeneity) in sperm head dimensions of individual stags may be a good indicator of sperm freezability.     

Treatment with eCG Decreases the Vascular Density and Increases the Glandular Density of the Bovine Uterus

The uterus plays an essential role in mammalian reproduction and is a target of several hormonal protocols used to improve fertility in cattle. Many studies highlighted the importance of eCG treatment following fixed‐time artificial insemination in improving follicular growth, ovulation and pregnancy rates in cattle. Moreover, eCG has been implicated in angiogenesis, leading to important changes in uterine blood flow and vascularisation. However, there is still a lack of information regarding the specific alterations induced by eCG upon glandular and vascular characteristics of bovine uterus. To investigate the influence of eCG on: uterine thickness and area; uterine artery diameter and area; uterine vascular and gland density; and the expression of the VEGFA‐system, the uteri of crossbred beef cows were collected. All cows were submitted to follicular wave emergence synchronization. On day four of protocol, cows submitted to superovulation (n = 6) received 2000 IU eCG, on day eight, after expected follicular deviation, cows submitted to stimulatory treatment (n = 5) received 400 IU eCG. Control cows (n = 5) did not receive eCG. On day five po cows were subjected to ultrassonographic evaluation and slaughtered for uterine tissue sampling on day six po. Uterine vessels and glands were quantified by the counting point stereological method. The VEGFA‐system was localized in different cellular types, showing no qualitative or quantitative differences in the site of expression or the intensity of the positive signal among the groups. Vascular density was decreased in the endometrium of stimulated and myometrium of superovulated cows compared with the control ones, which showed higher vascular density in the myometrium and endometrium of the ipsilateral uterine horn. The uterine gland density was higher in superovulated compared with stimulated and control cows. Thus, we can infer that stimulatory or superovulatory treatments with eCG influence the vascular density in the endometrium and myometrium in cattle.