Use of Powdered Egg Yolk vs Fresh Egg Yolk for the Cryopreservation of Ovine Semen

Egg yolk is a common additive to sperm cryopreservation diluents. Because of its animal origin, however, it also represents a potential risk of microbiological contamination in the diluent. This potential contamination can be avoided by using powdered egg yolk, instead of fresh egg yolk, as it is pasteurized. This study was conducted to determine ram sperm cryosurvival was affected by the type of egg yolk used (powdered egg yolk or fresh egg yolk) and by yolk concentration (10, 15 or 20%) in the diluent. Microbiological analyses were also performed to quantify the microbiological contamination in the diluents containing the two types of egg yolk. Sperm cryosurvival was determined by motility and morphology analyses after thawing. Motility parameters were assessed using a computer-assisted sperm analysis (CASA) system, and the percentage of sperm with a normal apical ridge was evaluated using a differential interference contrast microscope. No significant differences were observed between diluents in the percentage of sperm with normal apical ridge. However, higher percentages of total motile cells were observed for samples containing powdered egg yolk (69%) compared to samples containing fresh egg yolk (60%). However, sperm in diluents containing fresh egg yolk, exhibited higher values for average-path velocity, straight-line velocity and beat cross frequency and lower values for amplitude of lateral head displacement (p <0.05), compared to cells in diluents containing powdered egg yolk. Microbiological contamination was similar (<200 CFU/ml) in both diluents, and no bacterial growth was observed in either, when antibiotics were added. Therefore, powdered egg yolk can be effective used in diluents for the freezing of ram semen. However, the in vivo fertility of sperm frozen in diluents containing powdered egg yolk should be tested, as some motility parameters were different for sperm treated with powdered egg yolk compared to fresh egg yolk.     

Growth- and leaf-temperature effects on photosynthesis of sweet orange seedlings infected with Xylella fastidiosa

The effects of growth and leaf temperature on photosynthesis were evaluated in sweet orange seedlings ( Citrus sinensis cv. Pera) infected with Xylella fastidiosa (the bacterium that causes citrus variegated chlorosis, CVC). Measurements of leaf gas exchange and chlorophyll  a fluorescence were taken at leaf temperatures of 25, 30, 35 and 40°C in healthy and infected (without visible symptoms) seedlings submitted to two temperature regimes (25/20 or 35/20°C, day/night), not simultaneously. The CO₂ assimilation rates ( A ) and stomatal conductance ( g _s) were higher in healthy plants in both temperature regimes. Values for A and g _s of infected and healthy plants were higher in the 35/20°C regime, decreasing with leaf temperature increase. In addition, differences between healthy and infected plants were higher at 35/20°C, while no differences in chlorophyll  a fluorescence parameters were observed except for potential quantum efficiency of photosystem II, which was higher in infected plants. Low A values in infected plants were caused by low g _s and probably by biochemical damage to photosynthesis. The high alternative electron sink of infected plants was another effect of reduced A . Both high growth and high leaf temperatures increased differences in A between healthy and infected plants. Therefore this feature may be partially responsible for lower growth and/or productivity of CVC-affected plants in regions with high air temperature.     

Marine Cyclic Peptides: Antimicrobial Activity and Synthetic Strategies

Oceans are a rich source of structurally unique bioactive compounds from the perspective of potential therapeutic agents. Marine peptides are a particularly interesting group of secondary metabolites because of their chemistry and wide range of biological activities. Among them, cyclic peptides exhibit a broad spectrum of antimicrobial activities, including against bacteria, protozoa, fungi, and viruses. Moreover, there are several examples of marine cyclic peptides revealing interesting antimicrobial activities against numerous drug-resistant bacteria and fungi, making these compounds a very promising resource in the search for novel antimicrobial agents to revert multidrug-resistance. This review summarizes 174 marine cyclic peptides with antibacterial, antifungal, antiparasitic, or antiviral properties. These natural products were categorized according to their sources—sponges, mollusks, crustaceans, crabs, marine bacteria, and fungi—and chemical structure—cyclic peptides and depsipeptides. The antimicrobial activities, including against drug-resistant microorganisms, unusual structural characteristics, and hits more advanced in (pre)clinical studies, are highlighted. Nocathiacins I–III (91–93), unnarmicins A (114) and C (115), sclerotides A (160) and B (161), and plitidepsin (174) can be highlighted considering not only their high antimicrobial potency in vitro, but also for their promising in vivo results. Marine cyclic peptides are also interesting models for molecular modifications and/or total synthesis to obtain more potent compounds, with improved properties and in higher quantity. Solid-phase Fmoc- and Boc-protection chemistry is the major synthetic strategy to obtain marine cyclic peptides with antimicrobial properties, and key examples are presented guiding microbiologist and medicinal chemists to the discovery of new antimicrobial drug candidates from marine sources.     

Recent Advances in the Synthesis of Marine-Derived Alkaloids via Enzymatic Reactions

Alkaloids are a large and structurally diverse group of marine-derived natural products. Most marine-derived alkaloids are biologically active and show promising applications in modern (agro)chemical, pharmaceutical, and fine chemical industries. Different approaches have been established to access these marine-derived alkaloids. Among these employed methods, biotechnological approaches, namely, (chemo)enzymatic synthesis, have significant potential for playing a central role in alkaloid production on an industrial scale. In this review, we discuss research progress on marine-derived alkaloid synthesis via enzymatic reactions and note the advantages and disadvantages of their applications for industrial production, as well as green chemistry for marine natural product research.     

Comparison of duplex PCR and microscopic techniques for the identification of Babesia bigemina and Babesia bovis in engorged female ticks of Boophilus microplus

A single-step duplex polymerase chain reaction (PCR) technique and traditional microscopic examination of haemolymph smears were used to detect Babesia bigemina and/or Babesia bovis infection in engorged female ticks of Boophilus microplus recovered from calves raised in an endemic area of the State of Minas Gerais, Brazil. In the PCR amplification of tick-derived DNA, pairs of oligonucleotide primers specific for a 278-bp sequence from B. bigemina and for a 350-bp sequence from B. bovis were used conjointly. The microscopic examination of haemolymph revealed that 16.7% of the engorged ticks were infected with Babesia spp., although no significant differences (rho > 0.05) were found in the infection rate of ticks collected from calves of different age groups. PCR analysis showed that 77.8% of the engorged ticks whose haemolymph contained sporokinetes were infected with B. bigemina, 7.8% with B. bovis and 14.4% with both protozoan species. However, the PCR assay further revealed that, amongst the engorged female ticks whose haemolymph was apparently negative for the presence of sporokinetes, 15.6% were infected with B. bigemina, 2.2% with B. bovis and 10.0% with both species. The duplex PCR method is thus more efficient and sensitive than the microscopic assay and also permits facile identification of the protozoa species present in engorged female ticks.     

Habitat quality,not habitat amount,drives mammalian habitat use in the Brazilian Pantanal

Landscape Ecology - An understanding of species-habitat relationships is required to assess the impacts of habitat fragmentation and degradation. To date, habitat modeling in fragmented landscapes...     

Spatial patterns of the Citrus leprosis virus and its associated mite vector in systems without intervention

Leprosis is caused by the Citrus leprosis virus cytoplasmic type and is vectored by the mite Brevipalpus yothersi. Miticide applications, which cost $54 million annually, are based on inspection for the presence of mites. The aim of the present study was to characterize the spatial patterns of B. yothersi-infested trees and trees with leprosis symptoms for further improvement in sampling and disease control. The presence of mites and the occurrence of leprosis were assessed over two years in 1160 Valencia trees and 720 Natal trees in a commercial sweet orange grove in Sao Paulo State, Brazil. To assess the natural growth and dispersal of mites and leprosis, mite populations were not controlled during the experimental period. Maps of mite-infested trees and trees with leprosis symptoms were analysed at three different levels of spatial hierarchy using complementary methods, i.e. among adjacent trees within and across rows, within quadrats, and the strength and orientation of aggregation among quadrats. The study showed that the spatial patterns of virus-infected and mite-infested trees were different, with a strong aggregation pattern of trees with leprosis symptoms that increased over time. Conversely, the spatial pattern of B. yothersi showed randomness or weak aggregation at all three spatial hierarchical levels. Disease incidence increased steadily in plots of both cultivars, unlike in mite-infested trees where incidence fluctuated over time. These results have important implications for the development of better management strategies for leprosis. Sampling methods and action thresholds for mite control should consider primary disease inoculum in addition to the incidence of mites.     

Associations between morphometric variables and weight and yields carcass in Pirapitinga Piaractus brachypomus

The objective of this work was to evaluate the association between morphometric variables and carcass characteristics in Pirapitinga. We used a thousand specimens of Pirapitinga with an average weight of 1,200 g, which were stunned, slaughtered, weighed, measured, and processed for morphometric and processing yield analysis, to obtain weights, carcass and fillet yields. Initially, the linearity of the variables was verified. Pearson's simple and partial correlation tests were performed between all metrics. Track analysis was performed considering the weights and yields of carcass and filet as dependent variables and the others as independent variables. Ridge regression models were used to eliminate the effects of multicollinearity among the independent variables. Observations showed that the simple correlations between body weights and yields were superior to the partial ones in terms of magnitude. The length and circumference of the specimens were the measures most linearly associated with weight, carcass weight and fillet with rib weight. As for carcass yield and fillet yield, linear correlation coefficients were low and not significant when associated with body weights. With the path analysis, we could observe significant positive correlations between the morphometric measurements for weight, carcass weight and fillet with rib weight. The standard length, body circumference and body circumference/body height ratio are the most correlated. The latter are the most important variables in the evaluation of body weights in Pirapitinga fish. They can serve as criteria for indirect selection in searching for fish with better carcass characteristics. As for fillet and carcass yields, the morphometric variables have not shown to be efficient for indirect selection.