Cryopreservation of llama semen using a combination of permeable cryoprotectants

Semen cryopreservation is not available for massive use in South American Camelids (SACs) due to the lack of an efficient protocol and the low pregnancy rates obtained with artificial insemination (AI). The use of a single cryoprotectant (CP) is commonly used in SACs frozen semen. The objective of the study was to evaluate the combined cryoprotective capacity of two permeable CPs at different stages of the cryopreservation protocol in llama semen. Sixteen ejaculates from 4 llama males were analysed, and sperm quality was assayed in raw semen, at 5°C, after equilibration of samples with the CPs and when samples were thawed. The following CPs and combination were used: 6% glycerol (GL), 6% dimethylformamide (DMF) and the combination of both CPs: 3% GL and 3% DMF. A Kruskal–Wallis test and an experimental factorial design, considering one factor with four levels (raw semen, 6% GL, 6% DMF and GL/DMF), were used. Total sperm motility and live sperm with intact acrosomes remained unchanged after equilibration of samples (p > .05). A significant decrease in the percentage of functional membrane, motile and live sperm with intact acrosomes was observed when samples were thawed (GL, DMF and GL/DMF). Nevertheless, the cryopreservation protocols used preserved sperm DNA quality; thus, sperm chromatin condensation and DNA fragmentation were unaffected (p > .05) when GL, DMF and GL/DMF were used. To conclude, no superiority was found between the use of a single or a combination of permeable cryoprotectants to freeze llama semen.     

Influence of temperature and ontogeny on the levels of glucosinolates in broccoli (Brassica oleracea Var. italica) sprouts and their effect on the induction of mammalian phase 2 enzymes

Broccoli inflorescences have been recognized as components of healthy diets on the basis of their high content of fiber, vitamin C, carotenoids, and glucosinolates/isothiocyanates. Broccoli sprouts have been recently shown to have high levels of glucoraphanin (4-methylsulfinylbutyl glucosinolate), the precursor of the chemoprotective isothiocyanate, sulforaphane. This study evaluated the effects of temperature and developmental stage on the glucosinolate content of broccoli sprouts. Seedlings cultivated using a 30/15 degrees C (day/night) temperature regime had significantly higher glucosinolate levels (measured at six consecutive days postemergence) than did sprouts cultivated at lower temperatures (22/15 and 18/12 degrees C; p < 0.001). Both higher (33.1 degrees C) and lower (11.3 degrees C) constant temperatures induced higher glucosinolate levels in sprouts grown to a uniform size. Glucosinolate levels were highest in cotyledons and lowest in roots of sprouts dissected both early and late in the 11 day developmental span investigated. Nongerminated seeds have the highest glucosinolate levels and concordantly greater induction of mammalian phase 2 detoxication enzymes. Levels decline as sprouts germinate and develop, with consistently higher glucosinolate content in younger developmental stages, independent of the temperature regime. Temperature stress or its associated developmental anomalies induce higher glucosinolate levels, specific elevations in glucoraphanin content, and parallel induction of phase 2 chemoprotective enzymes.     

Angioarchitecture of collateral arteries of the aortic arch of Antillean manatee (Trichechus manatus manatus Linnaeus, 1758)

The aortic arrangement is an important structure associated with the maintenance of homeostasis. Based on this information, this study was conducted to describe the collateral arteries of the aortic arch of Antillean manatee and define the standard model for the species. Three specimens, an adult male, adult female and a male neonate, all strandings on the coast of the state of Rio Grande do Norte, Brazil, were used. The study was performed in the Laboratory of Morphophysiology of Vertebrates of the Federal University of Rio Grande do Norte, where in situ photographs were taken to demonstrate their topography. Subsequently, their hearts were removed and fixed in 10% formaldehyde and after 72 hr were dissected and analysed, obtaining schematic drawings and photographs of the vascular arrangement. The aortic arch was represented by three collateral arteries identified as the brachiocephalic trunk, left common carotid artery and left subclavian artery. This arrangement was similar to that found for other sirenians, and yet, for other mammals like hooded seal, murine, margarita island capuchin, black-handed tamarin, Mongolian gerbil and human. The morphological similarity presented in this study with different species of mammals, including humans, may contribute valuable information from an evolutionary point of view.     

Volatile Organic Compounds Profile Obtained from Processing Steps of Pacific Oysters (Crassostrea gigas) as Perspective for Food Industry

ABSTRACT

In the heat treatment of oysters, many aroma compounds are lost in the cooking steam and water. In this study, aroma compound profiles from oyster processing were identified by gas chromatography- mass spectrometry (GC-MS). The compounds were extracted by headspace-solid phase microextraction (HS-SPME), while the best process conditions such as temperature, extraction time, and desorption time were studied. The aroma compounds were obtained from raw oysters, from oysters after thermal treatment, from water released from raw oysters, from cooking steam, and from oyster boiling juice. All compounds were identified and semi-quantified by GC-MS. The major oyster aroma compounds were 1-octen-3-ol, hexanal, and benzaldehyde. Most of the components from oysters were lost both to the cooking steam and to the oyster boiling juice. The concentration range of volatile organic compounds transferred for oyster boiling juice between 14 and 90 mg L^?1 shows that the residues from oyster processing have great technological potential, and they can be recovered for application as aromas by the food industry.     

α‐Amylase Inhibitors from Wheat Against Development and Toxigenic Potential of Fusarium verticillioides

Fusarium verticillioides is one of the most important pathogens in maize and is a producer of fumonisin B₁ (FB₁). Although reports of its presence in wheat are scarce, the susceptibility of this cereal to fungus of the same genus motivates interest in investigating compounds present in the grain with inhibitory activity against this species. The aim of this study was to extract α‐amylase inhibitors from wheat and apply them in vitro to evaluate its effect on the development and expression of toxigenic potential of F. verticillioides. The α‐amylase inhibitors, both crude (P₀) and purified (P₁), were applied to in vitro culture containing a pathogen mycelium disc. Mycelial growth of the pathogen, glucosamine content, α‐amylase activity, and production of FB₁ were investigated. All protein extracts of wheat showed the ability to inhibit pathogen growth, especially the extract P₀ from cultivar Quartzo, which resulted in a reduction of glucosamine content (66%) and α‐amylase activity (84%). Furthermore, the protein inhibitors showed antifumonisin effect, reducing by 33 and 47% the mycotoxin production when applied as P₀ and P₁, respectively. These results suggest that α‐amylase inhibitor contributed to resistance against pathogen attack, acting in a diversified manner for each fungal species.     

Yield of synthetic backcross-derived lines in rainfed environments of Australia

Wheat is one of the major food crops in the world. It is Australia’s largest crop and most important agricultural commodity. In Australia the crop is grown under rainfed conditions with inherently important regional environmental differences; wheat growing areas are characterized by winter dominant rainfall in southern and western Australia and summer rainfall in northern Australia. Maximizing yield potential across these diverse regions is dependent upon managing, either genetically or agronomically, those factors in the environment that limit yield. The potential of synthetic backcross lines (SBLs) to increase yield in the diverse agroecological zones of Australia was investigated. Significant yield advantages were found for many of the SBLs across diverse environments. Depending on the environment, the yield of the SBLs ranged from 8% to 30% higher than the best local check in Australia. Apart from adaptation to semiarid water stressed conditions, some SBLs were also found to be significantly higher yielding under more optimal (irrigated) conditions. The four testing environments were classified into two groups, with the northern and southern environments being in separate groups. An elite group of SBLs was identified that exhibited broad adaptation across all diverse Australian environments included in this study. Other SBLs showed specific adaptation to either northern or southern Australia. This study showed that SBLs are likely to provide breeders with the opportunity to significantly improve wheat yield beyond what was previously possible in a number of diverse production environments.     

In vitro effect of Aloe vera, Coriandrum sativum and Ricinus communis fractions on Leishmania infantum and on murine monocytic cells

In South America, visceral leishmaniasis is a zoonosis caused by the protozoan species Leishmania infantum (syn. L. chagasi) and is primarily transmitted through the bite of the female Lutzomyia longipalpis. Its main reservoir in urban areas is the dog. The application of control measures recommended by health agencies have not achieved significant results in reducing the incidence of human cases, and the lack of effective drugs to treat dogs resulted in the prohibition of this course of action in Brazil. Therefore, it is necessary to search new alternatives for the treatment of canine and human visceral leishmaniasis. The objectives of this study were to evaluate the in vitro effect of fractions from Aloe vera (aloe), Coriandrum sativum (coriander), and Ricinus communis (castor) on promastigotes and amastigotes of L. infantum and to analyze the toxicity against the murine monocytic cells RAW 264.7. To determine the viability of these substances on 50% parasites (IC50), we used a tetrazolium dye (MTT) colorimetric assay (bromide 3-4.5-dimethylthiazol-2-yl-2,5-dephenyltetrazolium), and on amastigotes we performed an in situ ELISA. All fractions were effective against L. infantum promastigotes and did not differ from the positive control pentamidine (p>0.05). However, the R. communis ethyl acetate and chloroform fractions, as well as the C. sativum methanol fraction, were the most effective against amastigotes and did not differ from the positive control amphotericin B (p>0.05). The R. communis ethyl acetate fraction was the least toxic, presenting 83.5% viability of RAW 264.7 cells, which was similar to the results obtained with amphotericin B (p>0.05). Based on these results, we intend to undertake in vivo studies with R. communis ethyl acetate fractions due the high effectiveness against amastigotes and promastigotes of L. infantum and the low cytotoxicity towards murine monocytic cells.     

Acaricidal activity of thymol on larvae and nymphs of Amblyomma cajennense (Acari: Ixodidae)

The aim of the present study was to evaluate the acaricidal efficacy of thymol on unengorged and engorged larvae and engorged nymphs of Amblyomma cajennense. To perform the test for unengorged larvae, the larval packet technique was employed and mortality was evaluated 24h after the test, while for engorged larvae and engorged nymphs the immersion technique was employed and mortality was evaluated after 15 days. In all the experiments, the following concentrations of thymol were tested: 2.5, 5.0, 10.0, 15.0 and 20.0mg the thymol/ml. The control group was exposed to water and 1% DMSO and there were 10 repetitions for each treatment. The values found for mortality of unengorged larvae were 18.2%, 51.8%, 97.6%, 93.5% and 94.5%, and for engorged nymphs the values were 26.0%, 92.2%, 100.0%, 100.0%, 100.0%, in concentrations of 2.5, 5.0, 10.0, 15.0 and 20.0mg of thymol/ml, respectively. In the test with engorged larvae, mortality was 100.0% in all treatments. Based on the results, it is possible to conclude that thymol has acaricidal activity against immature stages of A. cajennense.     

Challenges of nematode control in ruminants: focus on Latin America

Gastrointestinal nematodes (GIN) of ruminants (cattle, sheep and goats) are ubiquitous and can cause severe injuries to infected animals and significant losses in farming revenues. GIN are able to survive severe environmental and host conditions, but mankind has developed a number of ingenious methods for parasite control. The commerce and use of modern anthelmintic drugs with a broad spectrum of activity has been a solid tool for nearly 40 years, however the continuous use of these drugs, has led to the selection of populations of drug-resistant worms worldwide. At present, the ever-growing agricultural systems in Latin America are facing many challenges and cannot rely on the far-reaching objective of parasitic elimination from the host or the environment. The lack of extensive programs for monitoring drug resistance exacerbates the negative consequences of reduced efficacy, which is evident in some areas with the increase in mortality rate even after treatment. Experts agree that new schemes of parasitic control are needed and should be based on the strategy of targeted selective treatment where affected hosts are identified and treated accordingly. In this article, we will focus our discussion on the challenges for the control of GIN in Latin America by 2020 imposed by reduced drug efficacy. We will evaluate phenotypic and molecular markers, methods for single-animal evaluation, and the implementation of schemes for anthelmintic treatment that address parasites in refugia.