Effects of different five-day progesterone-based synchronization protocols on the estrous response and follicular/luteal dynamics in dairy cows

This study compared the responses shown by lactating dairy cows to four different P4-based protocols for AI at estrus. Cows with no estrous signs 96 h after progesterone intravaginal device (PRID) removal were subjected to fixed-time AI (FTAI), and their data were also included in the study. In Experiment I, follicular/luteal and endometrial dynamics were assessed every 12 h from the beginning of treatment until AI. The estrous response was examined in Experiment II, and fertility was assessed in both experiments. The protocols consisted of a PRID fitted for five days, along with the administration of different combinations of gonadotropin releasing hormone (GnRH), equine chorionic gonadotropin and a single or double dose (24 h apart) of prostaglandin F_2α. In Experiment I (40 cows), animals receiving GnRH at the start of treatment showed a significantly higher ovulation rate during the PRID insertion period while estrus was delayed. In Experiment II (351 cows), according to the odds ratios, cows showing luteal activity at the time of treatment were less likely to show estrus than cows with no signs of luteal activity. Treatment affected the estrous response and the interval from PRID removal to estrus but did not affect conception rates 28–34 days post AI. Primiparous cows displayed a better estrous response than multiparous cows. Our findings reveal acceptable results of 5-day P4-based protocols for AI at estrus in high-producing dairy cows. Time from treatment to estrus emerged as a good guide for FTAI after a 5-day P4-based synchronization protocol.     

Insoluble fraction of buckwheat (Fagopyrum esculentum Moench) protein possessing cholesterol-binding properties that reduce micelle cholesterol solubility and uptake by Caco-2 cells

Buckwheat (Fagopyrum esculentum Moench) protein (BWP) exhibits hypocholesterolemic activity in several animal models by increasing fecal excretion of neutral and acidic sterols. In the current study, the ability of BWP to disrupt micelle cholesterol solubility by sequestration of cholesterol was investigated. When BWP (0.2%) was incubated with cholesterol and micelle lipid components prior to micelle formation, cholesterol solubility was reduced 40%. In contrast, cholesterol solubility was not decreased when BWP (0.2%) was incubated after micelle formation and incorporation of soluble cholesterol. Buckwheat flour, from which BWP was derived, had no significant effect on cholesterol solubility. Cholesterol uptake in Caco-2 cells from micelles made in the presence of BWP (0.2%) was reduced by 47, 36, 35, and 33% when compared with buckwheat flour, bovine serum albumin, casein, and gelatin, respectively. Reduction in cholesterol uptake in Caco-2 cells was dose-dependent, with maximum reductions at 0.1-0.4% BWP. In cholesterol-binding experiments, 83% of the cholesterol was associated with an insoluble BWP fraction, indicating strong cholesterol-binding capacity that disrupts solubility and uptake by Caco-2 cells.     

Detection of anti-Leishmania infantum antibodies in sylvatic lagomorphs from an epidemic area of Madrid using the indirect immunofluorescence antibody test

An outbreak of human leishmaniosis was confirmed in the southwest of the province of Madrid, Spain, between July 2009 and December 2012. Incidence of Leishmania infection in dogs was unchanged in this period, prompting a search for alternative sylvatic infection reservoirs. We evaluated exposure to Leishmania in serum samples from animals in the area with an indirect immunofluorescence test (IFAT). Using promastigotes from six culture passages and a 1/25 threshold titer, we found anti-Leishmania infantum seroreactivity in 9.3% of cats (4 of 43), 45.7% of rabbits (16/35) and 74.1% of hares (63/85). Use of promastigotes from >10 in vitro passages resulted in a notably IFAT lower titer, suggesting antigenic changes during extended culture. Postmortem inspection of seropositive animals showed no clinical signs of infection. The results clearly suggest that asymptomatic hares were the main reservoir in the outbreak, and corroborate IFAT as a sensitive serological surveillance method to detect such cryptic Leishmania infections.     

Radial stem growth in response to microclimate and soil moisture in a drought-prone mixed coniferous forest at an inner Alpine site

Dendroclimatological studies in a dry inner Alpine environment (750 m a.s.l.) revealed different growth response of co-occurring coniferous species to climate, which is assumed to be caused by a temporal shift in wood formation among species. The main focus of this study therefore was to monitor intra-annual dynamics of radial increment growth of mature deciduous and evergreen coniferous species (Pinus sylvestris, Larix decidua and Picea abies) during two consecutive years with contrasting climatic conditions. Radial stem growth was continuously followed by band dendrometers and modelled using Gompertz functions to determine time of maximum growth. Histological analyses of tree ring formation allowed determination of temporal dynamics of cambial activity and xylem cell development. Daily fluctuations in stem radius and radial stem increments were extracted from dendrometer traces, and correlations with environmental variables were performed. While a shift in temporal dynamics of radial growth onset and cessation was detected among co-occurring species, intra-annual radial growth peaked synchronously in late May 2011 and early June 2012. Moist atmospheric conditions, i.e. high relative air humidity, low vapour pressure deficit and low air temperature during the main growing period, favoured radial stem increment of all species. Soil water content and soil temperature were not significantly related to radial growth. Although a temporal shift in onset and cessation of wood formation was detected among species, synchronous culmination of radial growth indicates homogenous exogenous and/or endogenous control. The close coupling of radial growth to atmospheric conditions points to the importance of stem water status for intra-annual growth of drought-prone conifers.     

Genetic structure of Orbicella faveolata population reveals high connectivity among a marine protected area and Varadero Reef in the Colombian Caribbean

1. The protection of ecosystems with high diversity, such as coral reefs, is not an approach that guarantees their conservation. Thus, maintaining connectivity among coral populations over the long term is a strategy that should be adopted in order to protect diversity and ecological processes. Although coral reefs in Colombia are highly diverse, the population genetics baseline data of keystone species such as Orbicella faveolata are limited.
2. To provide current information about the connectivity between populations of O. faveolata in the Colombian Caribbean, the genetic diversity and distribution of O. faveolata was evaluated with analysis of molecular variance (AMOVA), principal components, and migration estimations using 113 genets distributed in six populations. Also, a genetic structure analysis that included the available data for the Caribbean population was conducted, seeking to understand how the Colombian populations relate to the broader region.
3. According to the global fixation index (F_ST) for the Colombian Caribbean, there was no genetic structure (F_ST = 0.002). Discriminant analysis of principal components (DAPC) showed that Corales del Rosario Archipelago (CR), Este de Isla Fuerte (EIF), San Bernardo Archipelago (SB), and Varadero Reef (VR) were grouped with Oeste de Isla Fuerte (OIF), with Bushnell (BS) representing the most divergent cluster. Migration analysis showed relatively high migration from VR to CR and SB, highlighting the importance of VR as a genetic reservoir for the region. Structure analysis showed that the Colombian population presented a specific genetic identity (F_ST = 0.254), suggesting that the Colombian Caribbean population could be a peripheral population that contributes significantly to genetic variation and is connected through a complex connectivity process.
4. In conclusion, the estimated genetic connectivity reflects the influence of sea surface dynamics over the interpopulation exchange dynamics and the role of protected and nonprotected coral reef in the Colombian Caribbean. Thus, the Colombian Caribbean population of O. faveolata could be relevant to the conservation of genetic diversity on a larger scale.

     

A multi-epitope chimeric protein elicited a strong antibody response and partial protection against Edwardsiella ictaluri in Nile tilapia

Edwardsiella ictaluri infects several fish species and protection of the all the susceptible fish hosts from the pathogen using a monovalent vaccine is impossible because the species is composed of host-based genotypes that are genetic, serological and antigenic heterogenous. Here, immunoinformatic approach was employed to design a cross-immunogenic chimeric EiCh protein containing multi-epitopes. The chimeric EiCh protein is composed of 11 B-cell epitopes and 7 major histocompatibility complex class II epitopes identified from E. ictaluri immunogenic proteins previously reported. The 49.32 kDa recombinant EiCh protein was expressed in vitro in Escherichia coli BL-21 (DE3) after which inclusion bodies were successfully solubilized and refolded. Ab initio protein modelling revealed secondary and tertiary structures. Secondary structure was confirmed by circular dichroism spectroscopy. Antigenicity of the chimeric EiCh protein was exhibited by strong reactivity with serum from striped catfish and Nile tilapia experimentally infected with E. ictaluri. Furthermore, immunogenicity of the chimeric EiCh protein was investigated in vivo in Nile tilapia juveniles and it was found that the protein could strongly induce production of specific antibodies conferring agglutination activity and partially protected Nile tilapia juveniles with a relative survival percentage (RPS) of 42%. This study explored immunoinformatics as reverse vaccinology approach in vaccine design for aquaculture to manage E. ictaluri infections.     

Selectivity in a trawl codend during haul-back operation—An overlooked phenomenon

The selectivity of a 99 mm trawl codend was assessed using a codend cover fitted with a MultiSampler, which was acoustically triggered to take separate samples at three different phases of the haul. The first sample was collected during towing, the second during haul-up and the third at the surface. A total of 18 hauls were conducted with a commercial fishing vessel west of Scotland. It was demonstrated that escapes take place not only during the tow but also in the short period when the trawl is hauled back from the seabed and when the codend is at the surface. For haddock (Melanogrammus aeglefinus), whiting (Merlangius merlangus) and Norway lobster (Nephrops norvegicus), respectively, the mean percentages escaping at the surface were 16, 12 and 38% of the total escape while 17, 8 and 28% escaped during the haul-up phase. Compared to towing, the escape rate (no./min) increased for haddock by a factor 2.7 during haul-up and by a factor 1.7 at the surface, whereas the escape rates of whiting were similar for the three phases. The escape rate of Norway lobster increased by a factor of approximately 7 for both the haul-up and surface phases, compared to the towing phase. The selectivity parameters L50 (50% retention length) and SR (selection range = L75–L25) were estimated and compared for the three different phases and for the whole haul for haddock, whiting and Norway lobster. For all three species there was no significant (P > 0.05) difference in L50 between the three phases of the haul. There was also no significant difference for whiting and Norway lobster when comparing the SR of the three phases, whereas the SR was significantly lower for haddock when comparing the surface phase with towing and haul-up. The estimate of L50 when towing was about 6 cm lower for haddock and whiting and 9 mm for Norway lobster compared to the selection curve estimated conventionally for the whole haul. Finally, the effect of sea state, duration and codend catch on the selectivity parameters were estimated for the individual phases and for the whole haul. A significant effect of at least one variable was found in all phases.     

Serodiagnosis of Lyme disease in UK dogs

Lyme disease is a chronic, multisystemic, inflammatory disorder of man and animals associated with infection by the tick-borne spirochaete, Borrelia burgdorferi. Lyme disease was recently reported for the first time in a dog in the UK (May and others 1990). Using an enzyme-linked immunosorbent assay (ELISA), we have performed a serological survey to investigate the prevalence of antibodies to B burgdorferi in UK dogs. The survey has shown that dogs from many areas in the UK have serum antibodies to B burgdorferi, that the presence of serum antibodies is associated with known exposure to ticks and that some dogs seropositive for B burgdorferi have clinical signs consistent with Lyme disease. High levels of serum anti-Borrelia antibodies are not diagnostic for canine Lyme disease, but, in association with appropriate clinical signs, they help to confirm the diagnosis in suspected cases.