Practical diets with essential oils of plants activate the complement system and alter the intestinal morphology of Nile tilapia

The effect of the essential oils (EOs) of peppermint, Mentha piperita L., and tea tree, Melaleuca alternifolia (Maiden & Betche) Cheel, on the haematological, biochemical, and immunological parameters and intestinal morphology of Nile tilapia, Oreochromis niloticus L., was evaluated. Fish (58.09 ± 5.87 g) were fed 100 mg/kg and 250 mg/kg of each EO and sampled on days 7, 14, 30 and 60 after starting supplementation. The haematological and biochemical parameters were not altered by the supplementation of EOs compared to the control (p > .05). With regard to the immunological parameters, the activation of the complement system of fish fed 250 mg/kg peppermint and 100 mg/kg and 250 mg/kg tea tree EOs were significantly higher compared to the control after 60 days of feeding (p < .05). The complement system plays an essential role in innate immunity and contributes significantly to the acquired immune response; thus, its activation through supplementation with EOs is promising for the formulation of nutritional additives in aquaculture. Regarding intestinal morphology, fish fed 250 mg/kg tea tree EO presented higher villus size compared to all other groups (p < .05), which represents a healthier gut. These fish present a larger intestinal surface, which can result in better absorption and utilization of the nutrients. Based on the responses found in this study, both EOs were considered promising for the formulation of feed additives for Nile tilapia.     

α-Rhamnosyl-β-glucosidase-catalyzed reactions for analysis and biotransformations of plant-based foods

Most aroma compounds exist in vegetal tissues as disaccharide conjugates, rutinose being an abundant sugar moiety in grapes. The availability of aroma precursors would facilitate analytical analysis of plant-based foods. The diglycosidase α-rhamnosyl-β-glucosidase from Acremonium sp. DSM 24697 efficiently transglycosylated the rutinose moiety from hesperidin to 2-phenylethanol, geraniol, and nerol in an aqueous-organic biphasic system. 2-Phenethyl rutinoside was synthesized up to millimolar level with an 80% conversion regarding the donor hesperidin. The hydrolysis of the synthesized aroma precursors was not detected in an aqueous medium. However, in the presence of ethanol as a sugar acceptor, the enzyme was able to transfer the disaccharide residue forming the alkyl-rutinoside. The aroma precursors were significantly hydrolyzed (up to 3-4% in 2 h at 30 °C), which indicated the potential use of the enzyme for biotechnological applications, for example, in aroma modulation of fermented foods.     

Cryptosporidium infection in a veal calf cohort in France: molecular characterization of species in a longitudinal study

ABSTRACT: Feces from 142 animals were collected on 15 farms in the region of Brittany, France. Each sample was directly collected from the rectum of the animal and identified with the ear tag number. Animals were sampled three times, at 5, 15 and 22 weeks of age. After DNA extraction from stool samples, nested PCR was performed to amplify partial 18S-rDNA and 60 kDa glycoprotein genes of Cryptosporidium. The parasite was detected on all farms. One hundred out of 142 calves (70.4%) were found to be parasitized by Cryptosporidium. Amplified fragments were sequenced for Cryptosporidium species identification and revealed the presence of C. parvum (43.8%), C. ryanae (28.5%), and C. bovis (27%). One animal was infected with Cryptosporidium ubiquitum. The prevalence of these species was related to the age of the animal. C. parvum caused 86.7% of Cryptosporidium infections in 5-week-old calves but only 1.7% in 15-week-old animals. The analysis of the results showed that animals could be infected successively by C. parvum, C. ryanae, and C. bovis for the study period. C. parvum gp60 genotyping identifies 6 IIa subtypes of which 74.5% were represented by IIaA15G2R1. This work confirms previous studies in other countries showing that zoonotic C. parvum is the dominant species seen in young calves.     

Intra-specific variability of virulence in Leishmania infantum zymodeme MON-1 strains

This study aims to characterize the intra-specific variability of virulence in Leishmania infantum zymodeme MON-1 strains isolated from dogs and immunocompetent and immunosuppressed patients through the evaluation of growth pattern, infective ability and immunopathogenicity. Two of the strains, classified as the most virulent, presented higher levels of macrophage infection, increased promastigote replication in culture medium and as well as amastigote multiplication within macrophages. These strains caused the most pathogenic infection inducing splenomegalia and maximum parasite loads in spleen and liver of BALB/c mice. The other strains exhibited either low virulence, with reduced infective capability and low replication levels, or an intermediate virulent phenotype showing mixed features similar to low and high virulent phenotypes. A correlation between the infectivity, growth dynamics and pathogenicity of each strain and the humoral and cellular immune response was demonstrated. Strains with accentuated virulent phenotype induced higher levels of anti-Leishmania IgG1 antibodies and TGF-beta but reduced production of IFN-gamma. Virulence phenotype seems to be a characteristic of each strain regardless of the host (dog or human) from which it was firstly isolated.     

Detection of <Emphasis Type="Italic">Trypanosoma vivax</Emphasis> DNA in semen from experimentally infected goats

The present work aimed to investigate the presence of T. vivax DNA in the semen of experimentally infected goats. Twelve male goats native to the Brazilian Northeast, adults, were randomly assigned to two experimental groups: the infected group consisting of six goats infected intravenously with 0.5 mL of blood containing approximately 1.25?×?10⁵ trypomastigotes of T. vivax, and a control group composed of six uninfected goats. After the infection, clinical examinations aiming to evaluate rectal temperature, parasitemia and hematocrit were performed. Semen samples were collected from goats by electroejaculation on the 7th, 14th and 21st days post-infection (dpi). The recombinant DNA-encoding gene encoding the L-like-specific gene for T. vivax. The infection was characterized by increased rectal temperature, high parasitemia and significant reduction of hematocrit values. Results for T. vivax DNA detection using TviCatL-PCR were positive in all semen samples from the infected group collected on 7th, 14th and 21st dpi. The presence of T. vivax DNA in 7th dpi suggests the early invasion of the parasite in the reproductive organs. Also, the finding of T. vivax DNA in all periods analyzed may suggest the continued elimination of the parasite in the semen, which may increase the chances of sexual transmission. Thus, T. vivax DNA is recorded for the first time in the semen of infected goats. Thus, these data are of great importance, since the detection of the T. vivax genetic material in the semen may point to the possibility that the parasite may be transmitted through the sexual pathway.     

Influence of seminal plasma during different stages of bovine sperm cryopreservation

This study aimed to evaluate the effect of seminal plasma on bovine sperm cryopreservation and to assess the integrity of plasma and acrosomal membranes, mitochondrial potential, remodelling of F-actin cytoskeleton and sperm chromatin fragmentation during the cooling, equilibrium and freezing/thawing stages. Six ejaculates collected from seven Nelore bulls (n = 42) were used in this study. Each ejaculate was divided into two aliquots (with seminal plasma = SP group; without seminal plasma = NSP group) and packed to a final concentration of 50 × 10⁶ sperm per straw. Statistical analyses were performed using SAS software (version 9.3), and p ≤ .05 was considered significant. A time effect was observed for all sperm characteristics (p < .05), except for chromatin fragmentation (p > .05). The presence of seminal plasma better preserved the acrosomal integrity (SP = 75.2% and NSP = 71.7%; p < .05) and also provided lower F-actin remodelling during cryopreservation process (SP = 29.9% and NSP = 32.4%; p < .05). Regarding to the cryopreservation stages, it was observed that cooling step induced higher remodelling of F-actin than the equilibrium and freezing/thawing stages (56.3%, 32.2% and 23.9%, respectively; p < .05). The equilibrium step had minor influence on overall sperm characteristics while the freezing/thawing stage was responsible for the highest percentage of damage in plasma membrane (−65.2%), acrosomal membrane (−34.0%) and mitochondrial potential (−48.1%). On the other hand, none of the cryopreservation stages affected chromatin integrity. It was concluded that the presence of seminal plasma provides increased acrosomal integrity and reduced remodelling of F-actin cytoskeleton. Higher F-actin remodelling is observed after the cooling step while the freezing/thawing step is most damaging to sperm membranes and mitochondrial potential during bovine sperm cryopreservation.     

Effect of the addition of diatoms (Navicula spp.) and rotifers (Brachionus plicatilis) on water quality and growth of the Litopenaeus vannamei postlarvae reared in a biofloc system

The aim of this study was to evaluate the effect of the addition of Navicula spp. and Brachionus plicatilis on water quality and growth of postlarvae shrimp Litopenaeus vannamei reared in a biofloc system. Four treatments were considered: a control (biofloc system – BFT); BFT with the addition of Navicula spp. (BFT‐N); BFT with the addition of Brachionus plicatilis (BFT‐B) and BFT with the addition of Navicula spp. and Brachionus plicatilis (BFT‐NB), each in triplicate. Shrimp (16.2 ± 0.03 mg) were stocked at a density of 2500 shrimp m^?3 and plankton were added on days 1, 5, 10, 15, 20, 25 and 30 at a density of 5 × 10⁴ cells mL^?1 (Navicula spp.) and 30 organisms L^?1 (Brachionus plicatilis). The shrimp were fed a formulated feed in four daily rations composed of 40% crude protein and 8% lipids. Significant differences between treatments were observed for final weight, yield, feed conversion ratio, specific growth rate, protein efficiency ratio and protein content of the shrimp. The combined plankton addition of Navicula spp. and B. plicatilis had better performance parameters, indicating their benefit as natural food sources for postlarvae L. vannamei in biofloc systems.     

Metabolic signature of spent culture media shows lipid metabolism as a determinant of pregnancy outcomes

In the present study, we investigated the spent culture media of in vitro produced (IVP) bovine embryos which did (group Pregnant) or did not (group Non-pregnant) establish pregnancy after transfer. For that purpose, IVP embryos on D5 were transferred to individual droplets for the last 48 h of culture. Embryos at the blastocyst stage were then transferred to synchronized recipients, while respective culture media drops were collected and evaluated individually. The list of metabolites present in spent culture media was obtained by electrospray ionization mass spectrometry (ESI-MS) and analysed with Metaboanalyst® to characterize the metabolic profile of each group. The spectrometric analysis showed that pathways related to lipid metabolism, particularly fatty acids degradation via beta-oxidation, were more present in the Pregnant group whereas no significant pathway was identified in the group Non-pregnant. By using this method, we were able to identify a metabolic signature in culture media that allows for a better comprehension of preferential metabolic routes taken by the most viable embryos. These findings offer great insights into the biochemistry of embryo development and reveal a potential target for the development of better-quality IVP systems, as well as tools to identify bovine embryos with greater chances to establish and maintain pregnancy.     

New Populations of Sclerotinia sclerotiorum from Lettuce in California and Peas and Lentils in Washington

ABSTRACT Four populations of Sclerotinia sclerotiorum in North America were inferred previously, based on analyses of both rapidly evolving markers (DNA fingerprint and mycelial compatiblity), and multilocus DNA sequence spanning the range between fast and slow evolution. Each population was defined as an interbreeding unit of conspecific individuals sharing a common recent ancestor and arising in a unique evolutionary event. The present study applies this standard to extend characterization of S. sclerotiorum populations to the Western United States. Isolates of S. sclerotiorum (N = 294) were determined to represent three genetically differentiated populations: California (CA, lettuce), Washington (WA, pea/lentil), and Ontario (ON, lettuce). CA was the most diverse population yet sampled in North America. Clonality was detected in ON and WA. No DNA fingerprints were common among the populations. The index of association (I(A)), based on fingerprint, was closer to zero (0) for CA than it was for the other populations. High diversity and lack of association of markers in California are consistent either with genetic exchange and recombination, or with large population size and high standing genetic variation. Intra- and interlocus conflict among three DNA sequence loci was consistent with recombination. The coalescent IGS genealogy confirmed subdivision and showed CA to be older than WA or ON. The Nearest Neighbor statistic on combined data confirmed subdivision among all present and previously defined populations. All isolates had both MAT1-1 and MAT1-2, consistent with uniform homothallism.