Serotyping and virulence genes detection in Escherichia coli isolated from fertile and infertile eggs, dead-in-shell embryos, and chickens with yolk sac infection

Escherichia coli is a common avian pathogen mainly associated with extraintestinal infections such as yolk sac infection (YSI). The aim of this study was to determine the serotypes and the presence of some virulence genes of E. coli strains isolated from different samples in a vertically integrated poultry operation in Mexico. Two hundred sixty-seven E. coli isolates from different samples were serotyped using rabbit serum against the 175 somatic (O) and 56 flagellar (H) antigens of the typing schema. Virulence genes were determined by colony blot hybridization, using DNA probes for st, eae, agg1, agg2, bfp, lt, cdt, slt, and ipaH diarrhea-associated virulence factors. The serogroup of 85% of the strains was determined; O19 (12%), 084 (9%), 08 (6%), and 078 (5%) were the most common. Using the complete antigenic formula (O and H), O19:NM (n = 31) was the serotype most frequently isolated from dead-in-shell embryos and in broilers that had died on the fourth, fifth, sixth, and seventh days after hatch. One hundred ten strains (41.2%) hybridized with one or more of the used probes. Of these, ipaH (72%), eae (30%), and cdt (27%) were the most common. Considering the origin of the respective isolates, 40% of the broiler farm strains were positive for at least one probe. Results show that some avian E. coli strains isolated in Mexico are included in avian pathogenic E. coli serotypes not previously reported, suggesting that they could be specific for this geographic area. The wide distribution of the ipaH gene among nonmotile strains suggests that this invasiveness trait could be important in YSI pathogenesis. On the other hand, some other genes could contribute to E. coli virulence during YSI.     

Morphology and SSR fingerprinting of newly developed <Emphasis Type="Italic">Cynara cardunculus</Emphasis> genotypes exploitable as ornamentals

The species Cynara cardunculus includes the globe artichoke (var. scolymus), the cultivated cardoon (var. altilis) and the wild cardoon (var. sylvestris). The three taxa are sexually compatible and originate fertile F ₁ progenies, which, given the high heterozygosity of the species, are highly segregating. We report the characterization of two F ₁ populations, one bred from a cross between globe artichoke and cultivated cardoon, and the other between globe artichoke and wild cardoon. Both populations featured a wide array of phenotypes in relation to several traits, and some of the newly developed genotypes are of interest for the ornamental market. The two populations were genotyped at 50 microsatellite (SSR) loci: in the globe artichoke × wild cardoon and globe artichoke × cultivated cardoon progenies 116 and 97 alleles were respectively detected. SSR pattern scores were used to produce an UPGMA dendrogram and a PCoA plot. A set of nine SSR loci, evenly dispersed across the genome, was shown to be sufficient to unambiguously identify each segregant. The molecular fingerprinting is useful for establishing the true to type correspondence of propagative materials in nurseries and ensures the effective correspondence between the real and the declared identity of a clone.     

Serum and tissue concentrations of doxycycline in broilers after the sub-cutaneous injection of a long-acting formulation

1. The antibacterial agent doxycycline hyclate (Dox) is usually administered to broilers in drinking water or as a feed supplement. Parenteral injection is not the usual route for administration, so a long-acting formulation (Dox-LA) was tested to evaluate if serum concentrations can achieve the pharmacokinetic/pharmacodynamic (PK/PD) ratios regarded as adequate for the drug.

2. A poloxamer-based matrix was used to provide Dox-LA. Serum and tissue concentrations of Dox vs time were determined in two day-old broilers after subcutaneous (SC) injection of Dox-LA or oral administration of a single bolus of aqueous Dox (Dox-PO), at a dose of 20?mg/kg. Weight gain, feed conversion rate, haematological variables, aspartate aminotransferase and alanine aminotransferase activities, blood urea and creatinine were determined and compared for Dox-LA with Dox-PO and non-medicated controls.

3. Dox-LA had a high relative bioavailability (1200%). Maximum serum concentrations were not statistically different (5·1?±?1·1?µg/ml for Dox-LA and 6·1?±?1.4?µg/ml for Dox-PO), but half-life of Dox-LA was much greater than the value obtained for Dox-PO (73·0?±?0·9?h and 2·0?±?0·02?h, respectively). Tissue concentrations were higher, and stayed higher for longer periods in the Dox-LA group.

4. In conclusion, considering the minimum effective serum concentration against Mycoplasma spp is 0·5?µg/ml, a dose-interval of 180?h can be achieved with Dox-LA, but only for 24?h after Dox-PO. Better PK/PD ratios for Dox-LA should result in improved clinical outcomes compared with Dox-PO.     

Novel Prunus rootstock somaclonal variants with divergent ability to tolerate waterlogging

Plants require access to free water for nutrient uptake, but excess water surrounding the roots can be injurious or even lethal because it blocks the transfer of free oxygen between the soil and the atmosphere. Genetic improvement efforts in this study were focused on the increased tolerance in roots to waterlogging. Among a pool of clones generated in vitro from leaf explants of rootstock Mr.S.2/5 of Prunus cerasifera L., the S.4 clone was flood tolerant whereas the S.1 clone was sensitive. The S.4 clone formed adventitious roots on exposure to flooding. Moreover, the chlorophyll content and mitochondrial activity in the leaf and root, soluble sugar content, alcohol dehydrogenase activity and ethylene content were different between the clones. The sorbitol transporter gene (SOT1) was up-regulated during hypoxia, the alcohol dehydrogenase genes (ADH1 and ADH3) were up-regulated in the leaves and down-regulated in the roots of the S.4 clone during hypoxia, and the 1-aminocyclopropane-1-oxidase gene (ACO1) was up-regulated in the leaves and roots of the S.4 clone during hypoxia and down-regulated in the wild-type roots. In addition, in the S.4 root, hypoxia induced significant down-regulation of a glycosyltransferase-like gene (GTL), which has a yet-undefined role. Although the relevant variation in the S.4 genome has yet to be determined, genetic alteration clearly conferred a flooding-tolerant phenotype. The isolation of novel somaclonals with the same genomic background but with divergent tolerance to flooding may offer new insights in the elucidation of the genetic machinery of resistance to flooding and aid in the selection of new Prunus rootstocks to be used in various adverse environments.     

Fatty Acid Profile,Total Carotenoids,and Free Radical-Scavenging from the Lipophilic Fractions of 12 Native Mexican Avocado Accessions

Plant Foods for Human Nutrition - Mexico is the world’s largest producer, exporter, and consumer of avocados. “Hass” avocado is the most commercialized cultivar, while the native...     

Influence of lipids in the generation of phenylacetaldehyde in wort-related model systems

The effect of lipids on the formation of the Strecker aldehyde phenylacetaldehyde during wort boiling was studied to determine the role that small changes in the lipid content of the wort have in the production of significant flavor compounds in beer. Wort was treated with 0-2.77 mmol per liter of glucose, linoleic acid, or 2,4-decadienal and heated at 60-98 degrees C for 1 h. After this time, the amount of the Strecker aldehyde phenylacetaldehyde increased in the samples treated with linoleic acid or decadienal but not in the samples treated with glucose. Thus, the amount of phenylacetaldehyde produced in the presence of linoleic acid was 1.1-2.5 times the amount of the Strecker aldehyde produced in the control wort, and this amount increased to 3.6-4.6 times when decadienal was employed. The higher reactivity of decadienal than linoleic acid for this reaction decreased with temperature and was related to the oxidation of linoleic acid that occurred to a higher extent at higher temperatures. The above results suggest that lipids can contribute to the formation of Strecker aldehydes during wort boiling and that changes in the lipid content of the wort will produce significant changes in the formation of Strecker aldehydes in addition to other well-known consequences in beer quality and yeast metabolism. On the other hand, because of the high glucose content in wort, small changes in its content are not expected to affect the amount of Strecker aldehydes produced.     

Diurnal and seasonal water variations of temperature,pH, redox potential and conductivity in gnammas (weathering pits): Implications for chemical weathering

A gnamma or weathering pit is a small-sized cavity sculpted into solid rock which refills with rainwater. This study describes the main hydrological processes in gnammas and the underlying chemical interactions with the environment. Four gnammas developed in a granite tor from central Spain were chosen for monitoring and studying the dynamics of the gnamma–water system. Parameters monitored were air and water temperature, pH, ORP (redox potential), and conductivity. These measurements describe the system dynamics, without the need for special sampling or other potential interferences under low water volumes. Additionally, chemical analyses were carried out to contrast the chemical properties with the chemical composition. As a consequence of water volumes less than 100 L, daily fluctuations tended to cause the largest amplitude changes. Photosynthesis and respiration/putrefaction processes in the gnamma are responsible for daily pH fluctuations up to 4 units, with pH values often greater than 9. ORP generally alternates from positive to negative values as the solution oscillates diurnally between basic and acidic conditions. Conductivity values ∼ 10 μS/cm occur most of the year, with only minor daily oscillations. During dry periods, conductivity increases significantly with maximum values > 400 μS/cm. These high conductivity values reflect atmospheric inputs by summer rainstorms after prolonged dry periods. Therefore, conductivity does not relate directly with chemical species dissolved from the host rock. Silica was the main weathering product of granite dissolution during the year, with highest concentrations (> 200 ppm) in the rainy season. Low Si values measured during the dry season are in accordance with observed precipitation of amorphous silica within the gnamma. Seasonal variations in concentration are also present in Al and Fe as a response to ORP variations and evaporative dynamics, causing mineral dissolution and new mineral precipitation during the year. Relations among the measured parameters are influenced by several factors, including biological processes, temperature effects and water level variations. Factors such as large pH oscillations, basic pH values, acids from biological activity, and alkali cations all increase silicate solubility, thus favoring granite dissolution. Overflow of water during the winter, when the water is chemically enriched, is an effective mechanism to remove weathering products.     

Seasonal and dietary effects on Vitamin D deficiencies detected in wild boar from mid‐western Spain

Vitamin D (VitD) is involved in important mammalian physiological mechanisms, such as Ca–P metabolism, bone development and immunological response. VitD deficiencies are frequently detected in domestic animals and related to various health problems (e.g., rickets, bone deformation). However, knowledge about the status of VitD in wildlife species, such as the wild boar, is scarce. The aims of this work were to explore VitD status in wild boar populations from mid‐western Spain and to elucidate the influence of daylight exposure and food supplementation in levels of VitD. Serum concentration of VitD (measured as 25‐hydroxivitaminD) was assessed in 276 wild boar from 27 game estates located in mid‐western Spain using a commercial ELISA kit. In 19 out of 27 estates, the staff supplied a specific VitD‐enriched food (2,000 UI/Kg) ad libitum throughout the year, while in the remaining estates (8), no food was supplied. Blood samples were extracted from hunted animals (198) between October and February of hunting seasons 2016/2017 and 2017/2018, and from live wild boar (78) that were captured, sampled and released (March–September of 2017). The percentage of animals with VitD deficiency (<20 ng/ml), VitD insufficiency (20–30 ng/ml) and VitD sufficiency (>30 ng/ml) was estimated, and the relationship of these levels to factors like sex, age and season was assessed using chi‐square tests. Furthermore, associations between daylight exposure and supplemental food with VitD levels were explored using linear models. Of the studied wild boar population, 82.2% showed a VitD deficiency or insufficiency. VitD deficiencies were more frequent in animals sampled in winter and spring. Furthermore, levels of VitD positively correlated with daylight exposure and supplemental food intake. Ad libitum supplementation with VitD‐enriched food was insufficient to prevent VitD deficiencies in wild boar from November to April, probably because food consumption is lower during this period.     

Low‐density lipoproteins and milk serum proteins improve the quality of stallion sperm after vitrification in straws

Lipids and proteins can be used for sperm vitrification to preserve the integrity of sperm membranes or to increase the viscosity of the medium. This study evaluated the effect of low‐density lipoproteins (LDL) and milk serum proteins (Pronexcell) for stallion sperm vitrification. Hippex extender (Barex Biochemical Products, The Netherlands), plus 1% of bovine serum albumin and 100 mM of trehalose, was used as control for sperm vitrification. In experiment 1, different concentrations of LDL (L1 = 0.25, L2 = 0.5, L3 = 1%) and in experiment 2 of Pronexcell (P1 = 1, P2 = 5, P3 = 10%) were added to control extender. Vitrification was performed in 0.25‐ml straws directly plunged into liquid nitrogen. Total motility (TM, %) and progressive motility (PM, %) were analysed by CASA, and plasma membrane (IMS, %) and acrosome membrane integrity (AIS, %) were assessed under epifluorescence microscopy. Post‐warmed sperm parameters were compared between treatments by ANOVA. Results were expressed as mean ± SEM. In both experiments, the minimum concentration of LDL and Pronexcell obtained significantly higher values (p < 0.01) than the control extender for TM (L1 = 52.95 ± 4.4; P1 = 58.99 ± 4.6; C = 30.88 ± 3.0), PM (L1 = 36.79 ± 5.5; P1 = 47.25 ± 4.3; C = 19.20 ± 2.4), IMS (L1 = 68.88 ± 3.6; P1 = 47.25 ± 4.3; C = 52.81 ± 2.6) and AIS (L1 = 45.88 ± 3.6; P1 = 47.25 ± 4.3; C = 26.00 ± 2.1). No differences in sperm parameters were found among different concentrations of LDL or Pronexcell. In conclusion, the addition of 0.25% LDL and 1% Pronexcell to the vitrification extender is recommended to improve the quality of stallion sperm after vitrification.