The citrus leafminer (CLM), Phyllocnistis citrella Stainton causes injury to citrus and related species in the Rutaceae family. The damage that the CLM larvae can cause is significant in citrus plantations. We tested two citrus cultivars — ‘Kinnow’ (Citrus reticulata Blanco) and ‘Fairchild’ (a hybrid of Citrus reticulata Clementine x Orlando Tangelo) — to quantify CLM larvae infestation and effect on the physiology of the citrus cultivars. We then compared the CLM larval weight with its associated damage. To calculate infestation level, mine area and total leaf area, we used the image analysis technique. The infestation level of CLM was higher in ‘Fairchild’ than in ‘Kinnow’ cultivar of citrus. For both cultivars, larval weight of CLM was directly proportional to the amount of mines generated. Taken together, the results of this study suggest that the mines that CLM larvae generate pose significant effect on the net photosynthetic rates and water use efficiency of citrus nursery plants. These results will help improve our understanding of the interaction between CLM and citrus nursery plants and effect of the pest on the yield potential of the crop.

This study investigated the potential application of genomic selection under a multi-breed scheme in the Spanish autochthonous beef cattle populations using a simulation study that replicates the structure of linkage disequilibrium obtained from a sample of 25 triplets of sire/dam/offspring per population and using the BovineHD Beadchip. Purebred and combined reference sets were used for the genomic evaluation and several scenarios of different genetic architecture of the trait were investigated. The single-breed evaluations yielded the highest within-breed accuracies. Across breed accuracies were found low but positive on average confirming the genetic connectedness between the populations. If the same genotyping effort is split in several populations, the accuracies were lower when compared with single-breed evaluation, but showed a small advantage over small-sized purebred reference sets over the accuracies of subsequent generations. Besides, the genetic architecture of the trait did not show any relevant effect on the accuracy with the exception of rare variants, which yielded slightly lower results and higher loss of predictive ability over the generations.

The aim of the study was to determine whether routine probiotic supplementation (RPS) with Lactobacillus rhamnosus GG (LGG) or Lactobacillus acidophilus +Lactobacillus bifidum is associated with reduced risk of necrotising enterocolitis (NEC)≥Stage II in preterm neonates born at ≤32 weeks’ gestation. We conducted a retrospective cohort study on the effect of probiotic supplementation in very low birth weight infants in our neonatal unit by comparing two periods: before and after supplementation. The incidence of NEC≥Stage II, late-onset sepsis and all-cause mortality was compared for an equal period ‘before’ (Period I) and ‘after’ (Period II) RPS with LGG or L. acidophillus+L. bifidum. Multivariate logistic regression analysis was conducted to adjust for relevant confounders. The study population was composed of 261 neonates (Period I v. II: 134 v. 127) with comparable gestation duration and birth weights. In <32 weeks, we observed a significant reduction in NEC≥Stage II (11·3 v. 4·8 %), late-onset sepsis (16 v. 10·5 %) and mortality (19·4 v. 2·3 %). The benefits in neonates aged ≤27 weeks did not reach statistical significance. RPS with LGG or L. acidophillus+L. bifidum is associated with a reduced risk of NEC≥Stage II, late-onset sepsis and mortality in preterm neonates born at ≤32 weeks’ gestation.

The Spanish local beef cattle breeds have most likely common origin followed by a process of differentiation. This particular historical evolution has most probably left detectable signatures in the genome. The objective of this study was to identify genomic regions associated with differentiation processes in seven Spanish autochthonous populations (Asturiana de los Valles (AV), Avileña-Negra Ibérica (ANI), Bruna dels Pirineus (BP), Morucha (Mo), Pirenaica (Pi), Retinta (Re) and Rubia Gallega (RG)). The BovineHD 777K BeadChip was used on 342 individuals (AV, n=50; ANI, n=48; BP, n=50; Mo, n=50; Pi, n=48; Re, n=48; RG, n=48) chosen to be as unrelated as possible. We calculated the fixation index (F ST ) and performed a Bayesian analysis named SelEstim. The output of both procedures was very similar, although the Bayesian analysis provided a richer inference and allowed us to calculate significance thresholds by generating a pseudo-observed data set from the estimated posterior distributions. We identified a very large number of genomic regions, but when a very restrictive significance threshold was applied these regions were reduced to only 10. Among them, four regions can be highlighted because they comprised a large number of single nucleotide polymorphisms and showed extremely high signals (Kullback–Leiber divergence (KLD)>6). They are located in BTA 2 (5 575 950 to 10 152 228 base pairs (bp)), BTA 5 (17 596 734 to 18 850 702 bp), BTA 6 (37 853 912 to 39 441 548 bp) and BTA 18 (13 345 515 to 15 243 838 bp) and harbor, among others, the MSTN (Myostatin), KIT-LG (KIT Ligand), LAP3 (leucine aminopeptidase 3), NAPCG (non-SMC condensing I complex, subunit G), LCORL (ligand dependent nuclear receptor corepressor-like) and MC1R (Melanocortin 1 receptor) genes. Knowledge on these genomic regions allows to identify potential targets of recent selection and helps to define potential candidate genes associated with traits of interest, such as coat color, muscle development, fertility, growth, carcass and immunological response.

This study aimed to evaluate the embryo development competence, the nuclear maturation and the viability of germinal vesicle (GV) and metaphase II (MII) oocytes vitrified by the Cryotop method. Cumulus–oocyte complexes were derived from bovine ovaries and three experiments were conducted. In Experiment 1, GV oocytes were vitrified and underwent in vitro maturation (IVM) or not and their nuclear maturation was assessed by orcein staining. In Experiment 2, GV oocytes and MII oocytes were vitrified or not and the viability was assessed by calcein/ethidium homodimer-1 staining. In Experiment 3, MII oocytes matured before or after vitrification were submitted to in vitro fertilization (IVF) and parthenogenetic activation (PA) in order to evaluate embryo development. No difference was found for the nuclear maturation rate in the GV group (50%) and the GV control group (67%; P = 0.23) and for viability rate (56%; 77%; P = 0.055, respectively). However, in the MII group (27%) viability was significantly lower than that of the MII control group (84%; P < 0.0001). The cleavage rate by IVF and PA was similar in the GV group and the MII group. In contrast, vitrified MII oocytes showed no capacity for blastocyst development after IVF or PA and vitrified GV oocytes were able to develop to blastocysts only after PA, but not after IVF. In conclusion, oocyte vitrification by the Cryotop method reduced the capacity for embryo development. Vitrification of GV oocytes, however, did not influence the capacity of meiotic nuclear maturation and they exhibited higher viability following vitrification at the MII stage.

The gastropod Buccinanops globulosus is commonly used as a bioindicator of tributyltin (TBT) contamination due to its high imposex incidence in maritime traffic areas. The aim of this study was to evaluate both oxidative stress in B. globulosus at three sites with different maritime activity, and imposex incidence in Nuevo Gulf, Argentina. Oxidative stress parameters in digestive glands, like superoxide dismutase (SOD) and glutathione-S-transferase (GST) activities, reduced glutathione levels (GSH), and oxidative damage to lipids, estimated as thiobarbituric acid reactive substances (TBARs) as well as imposex parameters (% imposex and female penis length (FPL)) were measured in females. Gastropods from the harbour area showed 100% imposex, the highest FPL and TBARs content, as well as GSH levels and SOD activity.

The different oxidative stress responses and high imposex incidence at the harbour site may indicate a negative effect on the organism's physiological state due to environmental pollution.

The distribution of phlebotomine sand flies is widely reported to be changing in Europe. This can be attributed to either the discovery of sand flies in areas where they were previously overlooked (generally following an outbreak of leishmaniasis or other sand fly-related disease) or to true expansion of their range as a result of climatic or environmental changes. Routine surveillance for phlebotomines in Europe is localized, and often one of the challenges for entomologists working in non-leishmaniasis endemic countries is the lack of knowledge on how to conduct, plan and execute sampling for phlebotomines, or how to adapt on-going sampling strategies for other haematophagous diptera. This review brings together published and unpublished expert knowledge on sampling strategies for European phlebotomines of public health concern in order to provide practical advice on: how to conduct surveys; the collection and interpretation of field data; suitable techniques for the preservation of specimens obtained by different sampling methods; molecular techniques used for species identification; and the pathogens associated with sand flies and their detection methods.

The mass mortality suffered by the sea urchin Diadema antillarum between 1983–1984 is considered one of the major causes of coral reef degradation in the Caribbean. Its near disappearance resulted in a disproportionate growth of macroalgae that has led to a ‘phase shift’ from coral-to-algal dominated reefs. The close relationship between this echinoid and the functioning of coral reef ecosystems makes it imperative to better understand the potential for recovery of its populations. From 2009 to 2011, we assessed the density and size structure of D. antillarum in various reefs where previous population data were available. Results indicate a modest increase in density in all localities with respect to the last time they were surveyed in 2003/2004. Nevertheless, density values are still lower than values reported for the island prior to the die-off. Overall density did not surpass 1.49 ind. per m−2, and did not change considerably during the studied period. Lack of population growth coincided with a lack of juveniles; suggesting that population growth at the studied sites may be limited by the number of individuals recruiting into the juvenile stage.

The long-spined sea urchin Diadema antillarum has been the focus of multiple studies since the mass mortality event in the 1980s. The recovery of this key herbivore in the wider Caribbean is essential for the well-being of coral reefs. This study examined the population density and structure of D. antillarum at seven northern fringing reefs of Puerto Rico between 2011 and 2013. The total mean density of the sea urchins in northern Puerto Rico was 0.9 ±0.3 ind m−2. Densities of D. antillarum significantly differed among sites, but not temporally. Differences in mean sizes were significant among sites and seasons. Areas with higher densities of D. antillarum showed lower cover of non-calcareous algae. Wave exposure was correlated with the abundance of the sea urchin. This study indicates that the observed abundance of D. antillarum has not yet returned to pre-mortality levels. However, densities showed some degree of recovery when compared with previous studies, enabling at least some degree of control on fleshy macroalgae communities. No significant changes in density occurred between 2011 and 2013, and sites with higher densities were generally located in leeward areas. The low relative abundance of small size individuals points towards recruitment limitation as an explanation for the limited recovery of D. antillarum.

Two in vitro experiments were conducted to analyse the effects of replacing dietary barley grain with wastes of tomato and cucumber fruits and a 1 : 1 tomato : cucumber mixture on rumen fermentation characteristics and microbial abundance. The control (CON) substrate contained 250 g/kg of barley grain on a dry matter (DM) basis, and another 15 substrates were formulated by replacing 50, 100, 150, 200 or 250 g of barley grain/kg with the same amount (DM basis) of tomato or cucumber fruits or 1 : 1 tomato : cucumber mixture. In Expt 1, all substrates were incubated in batch cultures with rumen micro-organisms from goats for 24 h. Increasing amounts of tomato, cucumber and the mixture of both fruits in the substrate increased final pH and gas production, without changes in final ammonia-nitrogen (NH3-N) concentrations, substrate degradability and total volatile fatty acid (VFA) production, indicating that there were no detrimental effects of any waste fruits on rumen fermentation. Therefore, in Expt 2 the substrates including 250 g of waste fruits (T250, C250 and M250 for tomato, cucumber and the mixture of both fruits, respectively) and the CON substrate were incubated in single-flow continuous-culture fermenters for 8 days. Total VFA production did not differ among substrates, but there were differences in VFA profile. Molar proportions of propionate, isobutyrate and isovalerate were lower and acetate : propionate ratio was greater for T250 compared with CON substrate. Fermentation of substrates containing cucumber (C250 and M250) resulted in lower proportions of acetate, isobutyrate and isovalerate and acetate : propionate ratio, but greater butyrate proportions than the CON substrate. Carbohydrate degradability and microbial N synthesis tended to be lower for substrates containing cucumber than for the CON substrate, but there were no differences between CON and T250 substrates. Abundance of total bacteria, Fibrobacter succinogenes and Ruminococcus flavefaciens, fungi, methanogenic archaea and protozoa were similar in fermenters fed T250 and CON substrates, but fermenters fed C250 and M250 substrates had lower abundances of R. flavefaciens, fungi and protozoa than those fed the CON substrate. Results indicated that tomato fruits could replace dietary barley grain up to 250 g/kg of substrate DM without noticeable effects on rumen fermentation and microbial populations, but the inclusion of cucumber fruits at 250 g/kg of substrate DM negatively affected some microbial populations as it tended to reduce microbial N synthesis and changed the VFA profile. More studies are needed to identify the dietary inclusion level of cucumber which produces no detrimental effects on rumen fermentation and microbial growth.

A survey of entomopathogenic nematodes (EPNs) belonging to the families Steinernematidae and Heterorhabditidae was conducted in three municipalities on the Pacific coast of the State of Colima, México, to determine their occurrence and recovery frequency and predominant plant species in cultivated and non-cultivated habitats. Nineteen soil samples were collected: seven from non-cultivated habitats and 12 from habitats or areas cultivated mostly with fruit and grain crops and grasses. Of the 19 soil samples, 14 were positive for EPNs; the total prevalence was 73.7%. From the 14 positive soil samples, 12 steinernematid isolates (85.7%) and two heterorhabditid isolates (14.3%) were recovered. Irrespective of the locations, EPNs from the genus Steinernema were recovered from the three municipalities; EPNs from the genera Steinernema and Heterorhabditis were recovered from Armería and Ixtlahuacán. Only steinernematid isolates were recovered from non-cultivated habitats. Most of the isolates were recovered from cultivated habitats, and our results suggest that there is a higher prevalence of EPNs in cultivated soils.

Two in vitro and one in vivo experiments were conducted to investigate the effects of a selection of plant compounds on rumen fermentation, microbial concentration and methane emissions in goats. Treatments were: control (no additive), carvacrol (CAR), cinnamaldehyde (CIN), eugenol (EUG), propyl propane thiosulfinate (PTS), propyl propane thiosulfonate (PTSO), diallyl disulfide (DDS), a mixture (40 : 60) of PTS and PTSO (PTS+PTSO), and bromochloromethane (BCM) as positive control with proven antimethanogenic effectiveness. Four doses (40, 80, 160 and 320 µl/l) of the different compounds were incubated in vitro for 24 h in diluted rumen fluid from goats using two diets differing in starch and protein source within the concentrate (Experiment 1).The total gas production was linearly decreased (P<0.012) by all compounds, with the exception of EUG and PTS+PTSO (P⩾0.366). Total volatile fatty-acid (VFA) concentration decreased (P⩽0.018) only with PTS, PTSO and CAR, whereas the acetate:propionate ratio decreased (P⩽0.002) with PTS, PTSO and BCM, and a tendency (P=0.064) was observed for DDS. On the basis of results from Experiment 1, two doses of PTS, CAR, CIN, BCM (160 and 320 µl/l), PTSO (40 and 160 µl/l) and DDS (80 and 320 µl/l) were further tested in vitro for 72 h (Experiment 2). The gas production kinetics were affected (P⩽0.045) by all compounds, and digested NDF (DNDF) after 72 h of incubation was only linearly decreased (P⩽0.004) by CAR and PTS. The addition of all compounds linearly decreased (P⩽0.009) methane production, although the greatest reductions were observed for PTS (up to 96%), DDS (62%) and BCM (95%). No diet–dose interaction was observed. To further test the results obtained in vitro, two groups of 16 adult non-pregnant goats were used to study in vivo the effect of adding PTS (50, 100 and 200 mg/l rumen content per day) and BCM (50, 100 and 160 mg/l rumen content per day) during the 9 days on methane emissions (Experiment 3). The addition of PTS and BCM resulted in linear reductions (33% and 64%, respectively, P⩽0.002) of methane production per unit of dry matter intake, which were lower than the maximum inhibition observed in vitro (87% and 96%, respectively). We conclude that applying the same doses in vivo as in vitro resulted in a proportional lower extent of methane decrease, and that PTS at 200 mg/l rumen content per day has the potential to reduce methane emissions in goats. Whether the reduction in methane emission observed in vivo persists over longer periods of treatments and improves feed conversion efficiency requires further research.

Nitrogen emissions from dairy cows can be readily decreased by lowering the dietary CP concentration. The main objective of this work was to test whether the milk protein yield reduction associated with low N intakes could be partially compensated for by modifying the dietary carbohydrate composition (CHO). The effects of CHO on digestion, milk N efficiency (milk N/N intake; MNE) and animal performance were studied in four Jersey cows fed 100% or 80% of the recommended protein requirements using a 4×4 Latin square design. Four iso-energetic diets were formulated to two different CHO sources (starch diets with starch content of 34.3% and NDF at 32.5%, and fiber diets with starch content of 5.5% and NDF at 49.1%) and two CP levels (Low=12.0% and Normal=16.5%). The apparent digestible organic matter intake (DOMI) and the protein supply (protein digestible in the small intestine; PDIE) were similar between starch and fiber diets. As planned, microbial N flow (MNF) to the duodenum, estimated from the urinary purine derivatives (PD) excretion, was similar between Low and Normal CP diets. However, the MNF and the efficiency of microbial synthesis (g of microbial N/kg apparently DOMI) were higher for starch v. fiber diets. Milk and milk N fractions (CP, true protein, non-protein N (NPN)) yield were higher for starch compared with fiber diets and for Normal v. Low CP diets. Fecal N excretion was similar across dietary treatments. Despite a higher milk N ouput with starch v. fiber diets, the CHO modified neither the urinary N excretion nor the milk urea-N (MUN) concentration. The milk protein yield relative to both N and PDIE intakes was improved with starch compared with fiber diets. Concentrations of β-hydroxybutyrate, urea and Glu increased and those of glucose and Ala decreased in plasma of cows fed starch v. fiber diets. On the other hand, plasma concentration of albumin, urea, insulin and His increased in cows fed Normal compared with Low CP diets. This study showed that decreasing the dietary CP proportion from 16.5% to 12.0% increases and decreases considerably the MNE and the urinary N excretion, respectively. Moreover, present results show that at similar digestible OM and PDIE intakes, diets rich in starch improves the MNE and could partially compensate for the negative effects of Low CP diets on milk protein yield.

The aims of this study were, first, to evaluate eye temperature (ET) with infrared thermography and heart rate (HR) to measure stress in horses during show jumping competitions and their relationship with competition results, and second, to evaluate the influence of different extrinsic and intrinsic factors of the horse on the stress measurements analysed. One hundred and seventy-three Spanish Sport Horses were analysed for ET and HR, and these measurements were taken 3 h before the competition, just after and 3 h after it. Two interval measurements were also assessed for each parameter. Positive significant correlations were found between ET and HR, measured before (r=0.23), just after competition (r=0.28) and for the later interval (r=0.26), whereas negative correlations with competition results were found only for ET when measured just after competing (r=−0.25). Two intrinsic factors (genetic line and age) and no extrinsic factors showed significant differences for ET, whereas one intrinsic factor (age) and two extrinsic factors (journey duration and number of training hours) showed significant differences for HR. The marginal means showed significantly higher ET values for the Anglo-Arab genetic line and for 5-year-old animals. HR values were significantly higher for 4-year-old animals, for horses which had travelled 4 to 6 h and for horses that had 3 to 6 h of daily training. This study suggests that, although ET and HR seemed to share a similar physiological basis, the factors that most influenced each parameter were different. Finally, ET seems to be a suitable tool for assessing stress during show jumping competitions in horses.