Postpartum Variations of Plasma IGF and IGFBPs,Oocyte Production and Quality in Dairy Cows: Relationships With Parity and Subsequent Fertility

The aim of this study was to determine whether postpartum variations of plasma IGF‐1 and IGFBP concentrations, oocyte production and quality were related to parity and subsequent conception rate in Holstein dairy cows. Holstein dairy cows [10 primiparous (PP) and 22 multiparous (MP)] were allotted in six batches and sampled once weekly between calving and oestrous synchronization treatment started at 71.2 ± 2.0 days postpartum. During the 3 weeks before treatment, ovum pick‐up (OPU) was performed twice weekly. Oocytes were scored on a 4‐point scale, and oocytes from OPU1, 3 and 5 were fertilized in vitro. Seventeen cows became pregnant after first and second AI and were considered as fertile (F), while the others were considered to be subfertile (SF). Logistic regression was carried out to investigate the relationships between repeated measurements and fertility including parity and batch effects in the models. Likelihood of fertility significantly increased when plasma urea and IGFBP‐3 concentrations decreased and was higher in PP compared with MP cows. There was a trend for fertility to increase when plasma IGF‐1 concentrations increased (p = 0.07). In vitro cleavage and development rates were similar between SF and F cows (46.4% and 28.3% in SF vs 55.0% and 22.1% in F). Parity had an effect on plasma IGF‐1 concentrations (PP: 61.65 ± 2.67 vs MP: 41.63 ± 5.81 ng/ml, p < 0.001), mean number of follicles aspirated per session (PP: 5.7 ± 1.3 vs MP: 9.5 ± 0.8, p < 0.05) and fertility (PP: 8/10 = 80% vs MP: 9/22 = 41%, p < 0.05) but not on the number of oocytes recovered per session nor their quality. In conclusion, postpartum plasma urea and IGFBP‐3 concentrations, but not oocyte production and quality before breeding, were related to subsequent conception rate in our experimental design. Parity had a significant effect on energy status, follicular growth and fertility and needs to be considered when investigating relationships between nutrition and reproduction.     

Genetic diversity and phylogenetic relationships between Leishmania infantum from dogs,humans and wildlife in south‐east Spain

Leishmania infantum causes human and canine leishmaniosis. The parasite, transmitted by phlebotomine sand flies, infects species other than dogs and people, including wildlife, although their role as reservoirs of infection remains unknown for most species. Molecular typing of parasites to investigate genetic variability and evolutionary proximity can help understand transmission cycles and designing control strategies. We investigated Leishmania DNA variability in kinetoplast (kDNA) and internal transcribed spacer 2 (ITS2) sequences in asymptomatically infected wildlife (n = 58) and symptomatically and asymptomatically infected humans (n = 38) and dogs (n = 15) from south‐east Spain, using single nucleotide polymorphisms (SNPs) and in silico restriction fragment length polymorphism (RFLP) analyses. All ITS2 sequences (n = 76) displayed a 99%–100% nucleotide identity with a L. infantum reference sequence, except one with a 98% identity to a reference Leishmania panamensis sequence, from an Ecuadorian patient. No heterogeneity was recorded in the 73 L. infantum ITS2 sequences except for one SNP in a human parasite sequence. In contrast, kDNA analysis of 44 L. infantum sequences revealed 11 SNP genotypes (nucleotide variability up to 4.3%) and four RFLP genotypes including B, F and newly described S and T genotypes. Genotype frequency was significantly greater in symptomatic compared to asymptomatic individuals. Both methods similarly grouped parasites as predominantly or exclusively found in humans, in dogs, in wildlife or in all three of them. Accordingly, the phylogenetic analysis of kDNA sequences revealed three main clusters, two as a paraphyletic human parasites clade and a third including dogs, people and wildlife parasites. Results suggest that Leishmania infantum genetics is complex even in small geographical areas and that, probably, several independent transmission cycles take place simultaneously including some connecting animals and humans. Investigating these transmission networks may be useful in understanding the transmission dynamics, infection risk and therefore in planning L. infantum control strategies.     

Chemical composition of Lippia spp. essential oil and antimicrobial activity against Aeromonas hydrophila

Phytotherapy can replace antibiotic administration as an alternative to control Aeromonas hydrophila, one of the main bacteria involved in the aetiology of farmed fish diseases. Given that plants of the Lippia spp. genus show biological potential for antimicrobial activity, this study evaluated the chemical composition of essential oils extracted from Lippia alba, Lippia origanoides and Lippia sidoides and their activity against A. hydrophila. The oils were obtained by steam distillation in a Clevenger‐type apparatus and their composition determined by gas chromatography and mass spectrometry (CG/MS). Antibacterial activity was assessed by calculating the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) using broth microdilution method. The main compounds identified were geranial (25.4%) and neral (16.6%) in L. alba oil, carvacrol (40.4%) and p‐cymene (11.4%) in L. origanoides oil and thymol (76.6%) and ortho‐cymene (6.3%) in L. sidoides oil. The three Lippia species showed bacteriostatic and bactericidal action against A. hydrophila, with MICs and MBCs ranging from 1250 to 5000 μg mL^?1. Of the species tested, the best performance was obtained with essential oil of L. sidoides.     

Dietary organic acids blend alone or in combination with an essential oil on the survival,growth, gut/liver structure and de hemato‐immunological in Nile tilapia Oreochromis niloticus

This study evaluated the effects of dietary supplementation of organic acids blend (OAB) alone or in combination with essential oil, Lippia origanoides (OAE) for Nile tilapia fed supplemented diets for 30 days. Fish (1.1 ± 0.04 g) were fed control (Control), or OAB 0.5% or OAB 0.5% + essential oil 0.125% (OAE) respectively. At the end of the experiment, samples were collected for de hemato‐immunological, histological analysis of the intestine and liver, as well as microbiology of the intestine. The pH of the diets supplemented with OAB and OAE reduced 0.92 and 0.19 respectively. The growth and FCR were unaffected by the treatments, but survival was significantly higher in the OAB treatment. Fish fed the OAB diet showed reduced concentration of total heterotrophic bacteria and Pseudomonas sp. in the intestine. Increased glucose in fish fed OAB and high number of circulating monocytes in fish fed OAE diet were observed. The anterior intestine of fish fed OAE diet showed larger number of goblet cells and increased villi height. The diet supplemented with OAB, mainly, improved the intestinal health and survival of tilapia juveniles and can be used in juvenile production.     

Lippia alba (Verbenaceae) hydrolate as sedative of tambaqui (Colossoma macropomum) juveniles in simulated transport conditions

Lippia alba hydrolate (LAH) is produced as a secondary product in essential oil extraction. The objective of this study was to evaluate the use of LAH in a simulation of the transport of tambaqui (Colossoma macropomum) juveniles. A series of 17 hr simulations were performed using load densities of 30, 60 and 90 juveniles per L and four LAH concentrations of 0.4%, 1%, 2% and 5%, with a control of just water. Fish survival, temperature, dissolved oxygen, conductivity, pH, ammonia excretion, alkalinity water hardness and net Na⁺, K⁺, Cl^‐ fluxes, were all evaluated before and after the transport simulations. Mucous cell densities in the gills were also determined microscopically, post simulation. The results showed that addition of 5% LAH achieved better survival during the simulation than the water control. Dissolved oxygen was significantly lower with an increased load density. Also, conductivity, NH₄ excretion and net ionic fluxes (K⁺ and Cl^?) were lower with 5% LAH, although mainly where higher load densities were tested. Mucous cell density was lower in LAH concentrations of 2% and 5%, again with higher load densities. In view of these findings, it is suggested that 5% LAH is an effective sedative for use in the transport of tambaqui, mainly with high load densities. This work is the first study of LAH and highlights its potential applications in aquaculture management.     

Chemical Characterization and Release of Polyphenols from Pecan Nut Shell [<Emphasis Type="Italic">Carya illinoinensis</Emphasis> (Wangenh) C. Koch] in Zein Microparticles for Bioactive Applications

The pecan nut [Carya illinoinensis (Wangenh) C. Koch] is a natural source of polyphenols with antioxidant properties. In this study, the encapsulation of aqueous and hydroalcoholic extracts of pecan nut shell were evaluated for the release of bioactive compounds and antioxidant potential in order to explore food applications using zein as encapsulating agent. The extracts showed high contents of total phenolics, condensed tannins and high antioxidant activity. Concentrations of proanthocyanidins were 9-fold higher in hydroalcoholic extracts. The LC-DAD analysis showed that catechins were the major phenolic compounds in samples, with epigallocatechin levels up to 138.62 mg mL^?1. Zein microparticles loaded with aqueous extract released 2.3 times more phenolic compounds than the hydroalcoholic extracts and the DSC thermograms showed that extracts of pecan nut shell remained thermally stable up to 240 °C. The zein microcapsules obtained in this study were efficiently encapsulated and represent an interesting additive due its high antioxidant capacity, physicochemical characteristics and morphology. The use of zein microparticles combined with natural extracts constitute a step forward in the improvement of current technology for delivering phenolic compounds with applications in functional foods and nutraceuticals.     

Environmental mastitis in intensive high‐producing dairy herds in New South Wales

Objective To determine the prevalence of mastitis pathogens in high‐producing intensive dairy herds in New South Wales. Design Field survey. Procedure Milk samples from the mastitis‐affected quarter were collected from cows on five high‐producing dairy farms in NSW. The 820 samples were cultured using standard microbiological culture techniques. Results Bacteria or fungi were isolated from 83.3% of samples (683/820). More than two colony types were isolated from 16.7% of samples (137/820), two types from 6.6% (54/820), and one type from 52.3% (429/820). No bacteria were isolated from 24.4% (200/820) of the primary cultures, but enrichment cultures of these samples yielded single colony type bacterial isolates from 36.5% (73/200) of samples. Environmental pathogens, including coliforms, environmental Streptococcus and Staphylococcus spp., made up 91% (555/610) of isolates and accounted for 33.6% (205/610), 41.6% (254/610) and 15.7% (96/610), respectively, of isolates. Escherichia coli accounted for 76.1% (156/205) of the coliform isolates, Streptococcus uberis and Streptococcus dysgalactiae accounted for 32.3% (82/254) and 28.0% (71/254), respectively, of the environmental streptococcal isolates. Contagious pathogens were uncommon, comprising only 2.5% (15/610) of the total isolates. Conclusion The incidence and causes of mastitis are largely influenced by farm management. The relatively high prevalence of coliform mastitis in the intensive high‐producing herds in this survey contrasts with the low incidence reported in surveys of pasture‐based herds in Victoria. If the Australian dairy industry continues its current trend of intensification, coliform intra‐mammary infections may emerge as an increasingly important cause of mastitis.     

Antioxidant, icthyotoxicity and brine shrimp lethality tests of Magonia glabrata

The ethanolic extract of the fruit bark from Magonia glabrata yielded shikimic acid, scopoletin, sitosterol glycoside and 2-O-methyl-l-inositol. Antioxidant, icthyotoxicity and brine shrimp lethality activities were observed in this extract. The major constituent, 2-O-methyl-l-inositol, was found to be inactive in two assays but showed moderate activity as a radical scavenger.     

Involvement of the β-cinnamomin elicitin in infection and colonisation of cork oak roots by <Emphasis Type="Italic">Phytophthora cinnamomi</Emphasis>

The virulence of two wild type (PA45 and PA37) and two genetically modified (13C: hygromycin resistant; FATSS: hygromycin resistant and β-cin knock-down) Phytophthora cinnamomi strains towards cork oak (Quercus suber) was assessed via a quantitative evaluation of disease symptoms arising from a soil infestation assay, and by a histological analysis of root colonization. Comparison of virulence, as expressed by symptom severity, resulted in the following ranking: highly virulent (wild type strains), medium virulence (strain 13C) and weakly virulent (FATSS). Both transgenic strains were compromised in their virulence, as expressed by symptom severity, but strain 13C was much less affected than FATSS. Microscopic observation showed that the FATSS strain was unable to effectively invade the root, while 13C and the two wild type strains were all able to rapidly colonize the whole root, including the vascular tissue. These results strengthen the notion that elicitins are associated, either directly or indirectly, with the infection process of Phytophthora.