Pathogen translocation and histopathological lesions in an experimental model of Salmonella Dublin infection in calves receiving lactic acid bacteria and lactose supplements

The purpose of this study was to evaluate the capacity of a lactic acid bacteria (LAB) inoculum to protect calves with or without lactose supplements against Salmonella Dublin infection by evaluating histopathological lesions and pathogen translocation. Fifteen calves were divided into three groups [control group (C-G), a group inoculated with LAB (LAB-G), and a group inoculated with LAB and given lactose supplements (L-LAB-G)] with five, six, and four animals, respectively. The inoculum, composed of Lactobacillus (L.) casei DSPV 318T, L. salivarius DSPV 315T, and Pediococcus acidilactici DSPV 006T, was administered with milk replacer. The LAB-G and L-LAB-G received a daily dose of 10⁹ CFU/kg body weight of each strain throughout the experiment. Lactose was provided to the L-LAB-G in doses of 100 g/day. Salmonella Dublin (2 × 10¹⁰ CFU) was orally administered to all animals on day 11 of the experiment. The microscopic lesion index values in target organs were 83%, 70%, and 64.3% (p < 0.05) for the C-G, LAB-G, and L-LAB-G, respectively. Administration of the probiotic inoculum was not fully effective against infection caused by Salmonella. Although probiotic treatment was unable to delay the arrival of pathogen to target organs, it was evident that the inoculum altered the response of animals against pathogen infection.     

Improved assessment of frozen/thawed mouse spermatozoa using fluorescence microscopy

Genetically modified (GM) animals are unique mutants with an enormous scientific potential. Cryopreservation of pre-implantation embryos or spermatozoa is a common approach for protecting these lines from being lost or to store them in a repository. A mutant line can be taken out of a breeding nucleus only if sufficient numbers of samples with an appropriate level of quality are cryopreserved. The quality of different donors within the same mouse line might be heterogeneous and the cryopreservation procedure might also be error-prone. However, only limited amounts of material are available for analysis. To improve the monitoring of frozen/thawed spermatozoa, commonly used in vitro fertilization (IVF) followed by embryo transfer were replaced with animal-free techniques. Major factors for assessing spermatozoa quality (i.e., density, viability, motility, and morphology) were evaluated by fluorescence microscopy. For this, a live/dead cell staining protocol requiring only small amounts of material was created. Membrane integrity was then examined as major parameter closely correlated with successful IVF. These complex analyses allow us to monitor frozen/thawed spermatozoa from GM mice using a relatively simple staining procedure. This approach leads to a reduction of animal experiments and contributes to the 3R principles (replacement, reduction and refinement of animal experiments).     

Evaluation of modified vaccinia virus Ankara expressing VP2 protein of infectious bursal disease virus as an immunogen in chickens

A recombinant modified vaccinia Ankara (MVA) virus expressing mature viral protein 2 (VP2) of the infectious bursal disease virus (IBDV) was constructed to develop MVA-based vaccines for poultry. We demonstrated that this recombinant virus was able to induce a specific immune response by observing the production of anti-IBDV-seroneutralizing antibodies in specific pathogen-free chickens. Besides, as the epitopes of VP2 responsible to induce IBDV-neutralizing antibodies are discontinuous, our results suggest that VP2 protein expressed from MVA-VP2 maintained the correct conformational structure. To our knowledge, this is the first report on the usefulness of MVA-based vectors for developing recombinant vaccines for poultry.     

The food contaminant fumonisin B1 reduces the maturation of porcine CD11R1+ intestinal antigen presenting cells and antigen-specific immune responses, leading to a prolonged intestinal ETEC infection

Consumption of food or feed contaminated with fumonisin B₁ (FB₁), a mycotoxin produced by Fusarium verticillioides, can lead to disease in humans and animals. The present study was conducted to examine the effect of FB₁ intake on the intestinal immune system. Piglets were used as a target and as a model species for humans since their gastro-intestinal tract is very similar. The animals were orally exposed to a low dose of FB₁ (1 mg/kg body weight FB₁) for 10 days which did not result in clinical signs. However, when compared to non-exposed animals, FB₁-exposed animals showed a longer shedding of F4⁺ enterotoxigenic Escherichia coli (ETEC) following infection and a lower induction of the antigen-specific immune response following oral immunization. Further analyses to elucidate the mechanisms behind these observations revealed a reduced intestinal expression of IL-12p40, an impaired function of intestinal antigen presenting cells (APC), with decreased upregulation of Major Histocompatibility Complex Class II molecule (MHC-II) and reduced T cell stimulatory capacity upon stimulation. Taken together, these results indicate an FB₁-mediated reduction of in vivo APC maturation.     

Changes of ochratoxin A in grapes inoculated with Aspergillus carbonarius and subjected to chamber-drying under controlled conditions

The production pattern for ochratoxin A in grapes inoculated with Aspergillus carbonarius and changes in its concentration during raisining of Merlot, Syrah, Tempranillo, and Cabernet Sauvignon red grapes and Pedro Ximenez white grape were studied. Grapes were chamber-dried under controlled temperature and humidity conditions, with and without dipping pretreatments in alkaline emulsions of olive oil or ethyl oleate. Based on the results for the grapes that developed the fungus (Merlot and Pedro Ximenez), a temperature of 50 °C in the absence of dipping stopped ochratoxin A production and even degraded part of the toxin already formed. Both dipping pretreatments facilitated removal of the toxin and led to its virtually complete disappearance. However, dipping in the ethyl oleate emulsion caused substantial changes in the sensory characters of the musts obtained from the raisins, so it should be avoided to ensure the expected quality in the sweet wines elaborated from them.     

Compost suppressiveness against Fusarium oxysporum was not reduced after one-year storage under various moisture and temperature conditions

The effect of storage conditions on compost suppressiveness against fusarium wilt of melon, caused by Fusarium oxysporum f. sp. melonis (FOM) was studied in relation to the dynamics of compost microbial activity and biodegradability. For this purpose, mature suppressive compost, prepared from tomato plants and separated cow manure, was divided into four portions and stored for one year under cool/warm (12 or 28 °C) or dry/wet (15-35 or 55-65% moisture content) conditions, in four different combinations: cool-dry, warm-dry, cool-wet and warm-wet. All composts retained and even enhanced their suppressive capacity during storage, with no significant differences among them by the end of the storage period. However, significant differences were found in the dynamics of some of the measured chemical and microbial properties. The microbial activity of composts stored under wet conditions was higher than that of those stored under dry condition, which resulted in a substantial decrease in dissolved organic matter content (expressed as dissolved organic carbon; DOC) and increase in its recalcitrance to biological degradation, decrease in basal heat emission, slower response to added glucose or citric acid, and higher NO₃ concentration, indicating increased nitrification under wet conditions. The DOC significantly correlated with several microbial properties as well as with compost suppressiveness of fusarium wilt of melon seedlings, and may be regarded as a most suitable general index for compost maturity. A best-subset multiple linear regression analysis revealed that the three best predictors, namely dissolved organic carbon (DOC), basal heat, and mesophilic bacterial counts, could explain as much as 83% of the total variance in compost suppressiveness. The generally agreed association between compost maturity and suppressiveness was verified in this case. It appears that compost microbial populations might compete and interfere with the saprophytic stage of FOM conidia, between germination and host invasion. In conclusion, it was demonstrated that compost suppressiveness against fusarium wilt of melon can be maintained for at least one year under a wide range of storage conditions, without any loss of suppressive capacity. This fact has positive logistical implications for the use of suppressive composts against FOM.     

Applying biofloc technology in the culture of juvenile of Piaractus brachypomus (Cuvier, 1818): Effects on zootechnical performance and water quality

The aim of this study was to assess the zootechnical performance and water quality of a cachama blanca (Piaractus brachypomus) culture using biofloc technology (BFT) versus a system with daily water exchange (DWE). To do this, 180 juveniles (mean initial weight: 5.40 ± 0.19 g) were distributed in 12 circular plastic tanks with stocking densities of 20 or 40 individuals m³; then, they were cultured for 91 days. BFT treatments kept a C:N ratio approximately of 15:1. Temperature, pH and oxygen were monitored daily, while the other variables were measured weekly. Most productive variables were significantly influenced by both culture system and stocking density with significantly higher values of daily weight gain, total weight gain and total length for fish kept in DWE 20. However, only minor differences were observed within the BFT system. With the exception of the toxic nitrogen compounds (NH₄⁺ and NO₂^?), all the other water quality parameters were within the acceptable ranges for the cultivation of tropical fish. Microorganisms started to settle from the first week. A total of 23 genera were present, the most outstanding of which being seven genera of ciliates and three rotifers, rhizopods and chlorophytes. In conclusion, both systems BFT and DWE are useful for increasing the production of P. brachypomus in captivity. Additionally, the BFT system can potentially be applied for growing juveniles of this specie in regions with scarce water resources.     

Activity,molecular mass and hydrolysis on baker's yeast protein of extracellular proteases from the putative probiotic bacteria Microbacterium sp. strain 8L and Exiguobacterium mexicanum strain 8N

The bacteria Microbacterium sp. 8L and Exiguobacterium mexicanum 8N are known to improve the culture of Artemia franciscana using baker's yeast as food. Using spectrophotometry, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS‐PAGE), substrate‐SDS‐PAGE and pH‐stat in vitro‐digestibility assays, the activity, molecular mass and hydrolysis on baker's yeast protein of proteases from extracellular polymeric substances (EPS) of the strains 8L and 8N along with the pathogenic strains Microbacterium sp. 8R and Vibrio parahaemolyticus 588 CECT (Vp) were studied. The EPSs of 8L and 8R showed one activity band, on which the serine inhibitor phenylmethylsulphonyl fluoride (PMSF) had no effect. The EPSs of 8N showed four bands; two were unaffected by PMSF, whereas one was affected, and the other was partially affected. The EPSs of Vp showed two bands, one partially inhibited by PMSF. No inhibitory effects from 1‐chloro‐3‐tosylamido‐7‐amino‐2‐heptanone (trypsin inhibitor) were observed in the protease bands of the studied bacteria. The EPSs of 8L and 8N showed a similar degree of hydrolysis (pH‐stat). The EPSs of 8L had the lowest Dice index of similarity of yeast protein profiles at 1 h of reaction. We conclude that the strain 8L could benefit A. franciscana by providing bacterial proteases for digestion of baker's yeast.     

Optimisation of Healthy-Lipid Content and Oxidative Stability during Oil Extraction from Squid (Illex argentinus) Viscera by Green Processing

Green extraction was applied to Argentinean shortfin squid (Illex argentinus) viscera, consisting of a wet pressing method including a drying step, mechanic pressing, centrifugation of the resulting slurry, and oil collection. To maximise the oil yield and ω3 fatty acid content and to minimise the oil damage degree, a response surface methodology (RSM) design was developed focused on the drying temperature (45–85 °C) and time (30–90 min). In general, an increase of the drying time and temperature provided an increase in the lipid yield recovery from the viscera. The strongest drying conditions showed a higher recovery than 50% when compared with the traditional chemical method. The docosahexaenoic and eicosapentaenoic acid contents in the extracted oil revealed scarce dependence on drying conditions, showing valuable ranges (149.2–166.5 and 88.7–102.4 g·kg⁻¹ oil, respectively). Furthermore, the values of free fatty acids, peroxides, conjugated dienes, and ω3/ω6 ratio did not show extensive differences by comparing oils obtained from the different drying conditions. Contrary, a polyene index (PI) decrease was detected with increasing drying time and temperature. The RSM analysis indicated that optimised drying time (41.3 min) and temperature (85 °C) conditions would lead to 74.73 g·kg⁻¹ (oil yield), 1.87 (PI), and 6.72 (peroxide value) scores, with a 0.67 desirability value.