Category Archives: ORIGINAL ARTICLE

Use of former food products in dairy buffalo nutrition: In vitro and in vivo evaluation

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Abstract

A feeding strategy that maintains high content of functional molecules in buffalo milk has been verified by giving Sorghum vulgare as green fodder, but it is not available all year round. The aim of this study was to evaluate the inclusion of former food products (FFPs) containing 87% biscuit meal (nonstructural carbohydrate: 60.1%; starch 14.7; crude protein 10.6), in the diet of buffaloes in terms of: (a) fermentation characteristics through gas production technique; (b) milk yield (MY) and quality; (c) content of some biomolecules and total antioxidant activity. The experiment was performed involving 50 buffaloes divided into two groups: Green group and FFPs group (animals fed Total Mixed Ration with either green forage or FFPs respectively). Daily MY was recorded and milk qualitative analyses were determined monthly for 90 days. Furthermore, fermentation characteristics of the diets were studied in vitro. No significant differences were recorded in feed intake, BCS and MY and quality. Similar in vitro fermentation data of two diets were found, with slight differences in terms of gas production and degradability. During the incubation, kinetic parameters showed a faster fermentation process with the diet of the FFPs group in relation to Green group (p < 0.05). Green group had higher levels (p < 0.01) of γ-butyrobetaine, glycine betaine, l-carnitine and propionyl l-carnitine in milk, whereas no differences were observed for δ-valerobetaine and acetyl l-carnitine. Total antioxidant capacity and iron reduction antioxidant assay were higher (p < 0.05) in the plasma and milk of the Green group. The administration of a diet high in simple sugars, obtained with FFPs, seems to favour the ruminal biosynthesis of some metabolites in milk, such as δ-valerobetaine and acetyl- l-carnitine, similar to green forage administration. Overall, the use of biscuit meal can be an alternative to green fodder when it is not available to ensure environmental sustainability and optimize costs without compromising milk quality.

Effect of Brazilian spinach (Alternanthera sissoo) pellet supplementation and dietary ratios on rumen characteristics, microorganisms, methane production, milk yield, and milk composition in dairy cows

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Abstract

The aim of the previous research was to evaluate the effects of Brazilian spinach pellet (BSP) supplementation and dietary ratios on rumen characteristics, methane estimation, and milk production in dairy cows. Four crossbred Thai dairy cattle, with Holstein Friesian (HF) cows with a body weight of 442 ± 50 kg were assessed in a 2 × 2 factorial in a 4 × 4 Latin square design to obtain diets; factor A was the roughage (R) to concentrate (C) ratio at 40:60 and 30:70, and factor B was level of BSP supplantation at 2% and 6% of dry matter (basis) intake (DMI). R:C ratio and supplementation of BSP had no interaction effect on DMI and nutrient digestibility. On DM, organic matter (OM), crude protein (CP), and acid detergent fiber (ADF) intake, the R:C ratio increased (p < 0.05). The digestibility of OM improved (p < 0.05) when cows were fed a R:C ratio of 30:70. On pH, ammonia-nitrogen, protozoal population, and blood urea-nitrogen, there were no interactions between the R:C ratio and BSP supplementation. Increasing the BSP supplementation to 6% (p < 0.01) decreased the protozoal population. The R:C ratio of 30:70 increased total volatile fatty acid (VFA) and propionate (C3) concentrations while decreasing the acetate (C2) to C3 ratio and methane (CH4) estimation (p < 0.01). The average concentration of total VFA has increased by 114.46 mmol/L for 6% of BSP supplementation. Increased BSP supplementation increased the C3 concentration while decreasing the C2:C3 ratio and CH4 emissions (p < 0.05). The R:C ratio and BSP supplementation had no interaction effect on milk yield, 3.5% fat-corrected milk (FCM), or milk composition. The R:C ratio of 30:70 increased milk yield (p < 0.05) to the highest level of 12.18 kg/day. In conclusion, the diet containing a R:C ratio of 30:70 increased feed intake, milk yield, BUN, total VFA, and C3 concentration, and decreased the C2:C3 ratio and CH4 emission. BSP supplementation at 6% could increase TVFA and C3 concentrations while decreasing the protozoal population and CH4 estimation.

Low water supply differentially affects the growth, yield and mineral profile of kabuli and desi chickpeas (Cicer arietinum)

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Low water supply differentially affects the growth, yield and mineral profile of kabuli and desi chickpeas (Cicer arietinum)

At low water supplies, kabuli showed higher shoot water content, while desi had higher root water content. Shoot length was higher in kabuli, while root length increased in both genotypes. The root/shoot ratio was higher in kabuli and increased with water stress, negatively affecting pod and seed production. Lipid peroxidation also increased with water stress in kabuli, having positive correlations with growth parameters but negative with productivity. Water supply did not affect seed mineral accumulation, but desi sustained higher levels of P, Mg, Zn, Fe, Mn and B than kabuli, including at lower water supplies.


Abstract

The climatic events predicted to increase in intensity and frequency in the near future, including drought, may influence the quality and productivity of several important crops for human nutrition, such as legumes. Herein, two chickpea genotypes (Cicer arietinum) were analysed for their resilience to low water supply: a commercial white chickpea (kabuli) and a traditional black chickpea (desi) with marginal production in occidental countries. Plants were grown under four levels of water supplies (90%, 75%, 50% and 25% of field capacity) and biometric variables (root, shoot, pods and seeds), proxies of plant fitness (water content and oxidative stress) and the seed nutritional profile (protein and mineral concentrations) were analysed at plant maturity. The results show that the water content in shoots and roots decreased with the decrease in water supplies, with kabuli plants generally having higher water content in shoots and desi in roots. The shoot length was significantly higher in kabuli compared to desi, while the root length increased up to 11% in both species with the decrease in water supplies. The root-to-shoot ratio was higher in kabuli and increased with the decrease in the water supply, being negatively correlated with the number of pods and seeds per plant. Lipid peroxidation also increased with the decrease in the water supply, having slight positive correlations with plant growth parameters while being negatively correlated with plant productivity. No significant effects of plant genotype and water supply were observed on seed K, Ca and protein, but desi was able to sustain higher P, Mg, Zn, Fe, Mn and B concentrations than kabuli, including at lower water supplies. The results suggest that water stress negatively impacts plant growth and productivity and that the two chickpea genotypes have distinct biomass and water allocation strategies to cope with low water supply. These findings may be useful in strategies for improving the productivity and nutritional profile of chickpea crops under water-limited conditions.

The quality of beef in the conditions of the former Semipalatinsk Test Site

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Abstract

The territory of the Semipalatinsk Nuclear Test Site represents vast areas of grassy steppes, their use as pastures seems promising. The purpose of work was to compare beef samples obtained from settlements belonging to different categories of radiation risk: Kokpekty village, Chagan urban-type settlement, Krivinka village, Sarzhal village, three samples were examined from each settlement. Organoleptic analysis, radiometric determination of 137Cs, was used as criteria. Also, an analysis of the amino acid composition by the high-performance liquid chromatograph method was carried out and a calculation of protein indicators was made: meat tenderness, amino acid usefulness, nutrition value. The organoleptic examination showed the compliance of all samples with the standards, with the exception of one sample from the Sarzhal village. The study of the specific activity of 137Cs showed a direct correlation between the category of radiation risk and the content of this radioisotope in meat from different zones. The availability of some deviations in the samples indicators obtained from the Sarzhal village indicates the need to approach the products control from this zone more carefully, and radioisotope analysis should become a decisive criterion in determining the safety profile of the product.

mTORC2‐AKT‐LAT1 signalling participates in methionine‐induced β‐CASEIN expression in mammary epithelial cells of dairy cows

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Abstract

This study investigated the role of the mammalian target of rapamycin complex 2 (mTORC2)-protein kinase B (AKT) signalling in methionine (Met)-induced L-type amino acid transporter 1 (LAT1) expression and milk protein production. Primary mammary epithelial cells (MECs) from mammary parenchymal tissues of three lactating cows and MAC-T bovine MECs were cultured with or without 0.6 mM Met. Rapamycin-insensitive companion of mTOR (RICTOR) siRNA, the mTORC1 inhibitor rapamycin and the AKT activator SC79 were used to evaluate the effects of mTORC2-AKT signalling on Met-induced LAT1 expression and function. Each experiment was performed three times. Data were analysed with a two-sided unpaired t test or ANOVA with the Bonferroni multiple-comparison test. Western blotting showed that Met stimulation increased RICTOR expression (~244.67%; p < 0.05; control, 0.15 ± 0.026; Met, 0.517 ± 0.109) and AKT-S473 levels (~281.42%; p < 0.01; control, 0.253 ± 0.067; Met, 0.965 ± 0.019) in both primary MECs and MAC-T cells. Rapamycin-induced mTORC1 signalling inhibition decreased only Met-induced β-CASEIN expression by ~21.24% (p < 0.01; Met, 0.777 ± 0.01; Met and rapamycin, 0.612 ± 0.04) and did not affect Met-stimulated AKT-S473 levels, suggesting that mTORC2-AKT activation upon Met stimulation also contributes to milk protein synthesis. LAT1 participates in Met-induced β-CASEIN expression. In dairy cow MECs, mTORC2 inhibition by RICTOR siRNA decreased LAT1 levels on the plasma membrane by ~45.13% (p < 0.01; control, 0.359 ± 0.006; siRICTOR, 0.197 ± 0.004). However, SC79-induced AKT activation had the opposite effect (p < 0.01). In primary MECs and MAC-T cells, Met stimulation increased cytosolic and plasma membrane LAT1 expression respectively (MECs, 113.98% and 58.43%; MAC-T, 165.85% and 396.39%; p < 0.05). However, RICTOR siRNA significantly reduced Met-induced plasma membrane LAT1 expression (~76.48%; Met, 0.539 ± 0.05; Met and siRICTOR, 0.127 ± 0.012; p < 0.05). Thus, Met increased LAT1 expression and function via mTORC2-AKT signalling, upregulating milk protein synthesis in dairy cow MECs.

In vitro aflatoxins recovery after changing buffer or protozoa concentrations in the rumen fermentation fluid

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Abstract

This study simulates in vitro the effects of (i) rumen acidity and (ii) change in rumen protozoa numbers on the recovery of aflatoxins (AFs). Two 24-h fermentation experiments were carried out using the same batch in vitro fermentation systems and substrate (dried corn meal) containing 11.42, 2.42, 7.65 and 1.70 µg/kg of AFB1, AFB2, AFG1 and AFG2 respectively. In Experiment 1, two buffer concentrations (normal salts dosage or lowered to 25%) were tested. Buffer reduction decreased gas production (730 vs. 1101 mL, p < 0.05), volatile fatty acids (VFA) and NH3 concentrations in the fermentation liquid (39.8 vs. 46.3 mmol/L, and 31.7 vs. 46.5 mg/dL respectively, p < 0.01). Recovery of all four AFs types was higher (p < 0.01) in the reduced buffer fermentation fluid, both as a percentage of total AF incubated (73.6% vs. 62.5%, 45.9% vs. 38.1%, 33.6% vs. 17.9% and 18.9% vs. 6.24% for AFB1, AFB2, AFG1 and AFG2 respectively) and as amounts relative to VFA production (163.4 vs. 123.5, 22.1 vs. 15.7, 48.8 vs. 22.5 and 6.16 vs. 1.86 ng/100 mmol of VFA, for AFB1, AFB2, AFG1 and AFG2 respectively). In Experiment 2, Stevia rebaudiana Bertoni extracts (S) or a Camphor essential oil (Cam) were added to fermenters and compared to the control (no additives, C). S and Cam addition resulted in a 25% reduction (p < 0.05) and a 15% increase (p < 0.05) in protozoa counts respectively, when compared to C. Both plant additives slightly reduced (p < 0.05) AFB1 recovery as a percentage of total AFB1 incubated (68.5% and 67.7% vs. 74.9% for S, Cam and C respectively). Recoveries of all other AFs were unaffected by the additives. In conclusion, the rumen in vitro AFB1 recovery (63%–75%) was higher than other AFs (3%–46%) and the acidic fermentation environment increased it. In our conditions, changes in protozoa numbers did not affect AFs recovery.

Transfer of Phi6 bacteriophage between human skin and surfaces common to consumer‐facing environments

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Abstract

Aims

This study aimed to determine the extent of Phi6 (Φ6) transfer between skin and surfaces relevant to consumer-facing environments based on inoculum matrix, surface type and contact time.

Methods and Results

Φ6 transfer rates were determined from skin-to-fomite and fomite-to-skin influenced by inoculum matrix (artificial saliva and tripartite), surface type (aluminium, plastic, stainless steel, touchscreen, vinyl and wood) and contact time (5 and 10 s). Significant differences in estimated means were observed based on surface type (both transfer directions), inoculum matrix (skin-to-fomite) and contact time (both transfer directions). During a sequential transfer experiment from fomite-to-skin, the maximum number of consecutive transfer events observed was 3.33 ± 1.19, 2.33 ± 1.20 and 1.67 ± 1.21 for plastic, touchscreen and vinyl, respectively.

Conclusions

Contact time significantly impacted Φ6 transfer rates, which may be attributed to skin absorption dynamics. Surface type should be considered for assessing Φ6 transfer rates.

Significance and Impact of the Study

Although the persistence of Φ6 on fomites has been characterized, limited data are available regarding the transfer of Φ6 among skin and fomites. Determining Φ6 transfer rates for surfaces in consumer-facing environments based on these factors is needed to better inform future virus transmission mitigation strategies.

Rapid phylogenetic analysis using open reading frame content patterns acquired by Oxford nanopore sequencing

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Abstract

Aims

Phylogenetic analysis based on core genome single nucleotide polymorphisms (cgSNPs) using whole-genome sequencing (WGS) is increasingly used in epidemiological investigations of bacteria. The approach, however, is both resource intensive and time-consuming. Oxford Nanopore Technologies (ONT) sequencing is capable of real-time data analysis but the high error rate hampers its application in cgSNP-based phylogenetic analysis. Here, we developed a cgSNP-independent phylogenetic analysis method using ONT read assemblies by focusing on open reading frame (ORF) content patterns.

Methods and Results

WGS data of 66 Enterobacter hormaechei strains acquired by both ONT and Illumina sequencing and 162 strains obtained from NCBI database were converted to binary sequences based on the presence or absence of ORFs using BLASTn. Phylogenetic trees calculated from binary sequences (ORF trees) were compared with cgSNP trees derived from Illumina sequences. Clusters of closely related strains in the cgSNP trees formed comparable clusters in the ORF trees built with binary sequences, and the tree topologies between them were similar based on Fowlkes–Mallows index.

Conclusions

The ORF-based phylogenetic analysis using ONT sequencing may be useful in epidemiological investigations and offer advantages over the cgSNP-based approach.

Significance and Impact of the study

Conversion of assembled WGS data to binary sequences based on the presence or absence of ORFs circumvents read error concerns with ONT sequencing. Since ONT sequencing generates data in real time and does not require major investment, this ORF-based phylogenetic analysis method has the potential to enable phylogenetic and epidemiological analysis at the point of care.

A comparative genomics approach for identifying genetic factors in Escherichia coli isolates associated with bovine diseases

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Abstract

Aims

E. coli are ubiquitously present bacterial pathogens that cause septicaemia, diarrhoea and other clinical illness in farm animals. Many pathogen factors can be associated with disease conditions. Currently, studies inferring E. coli genetic factors associated with infection in bovines are limited. Hence, the present study envisaged to determine the pathogen genetic factors associated with bovine disease conditions.

Method and Results

The comparative genomic analysis involved genome sequence data of 135 diseased and 145 healthy bovine origin E. coli strains. Phylogroups A and C, as well as pathotypes ExPEC and EPEC, were found to have a strong connection with bovine disease strains. STEC strains, including EHEC, seem to play a less important role in bovine disease. Sequence types (STs) predominant among strains from diarrhoeal origin were ST 301 (CC 165) and ST 342. Correlation of core genome phylogeny with accessory gene-based clustering, phylogroups and pathotypes indicated lineage-specific virulence factors mostly associated with disease conditions.

Conclusions

Comparative genomic analysis was applied to infer genetic factors significant in bovine disease origin E. coli strains. Isolates from bovine disease origin were enriched for the phylogroups A and C, and for the pathotypes ExPEC and EPEC. However, there was minimal evidence of STEC involvement. The study also indicated predominant genetic lineages and virulence genes (pap, sfa and afa) associated with disease origin strains.

Significance and impact of study

The study revealed significant pathotypes, phylogroups, serotypes and sequence types associated with bovine disease conditions. These identified genetic factors can be applied for disease diagnosis, implementing vaccines and therapeutic measures. In addition, E. coli isolates from the bovine species revealed a complex pattern of disease epidemiology.

Limosilactobacillus fermentum‐fermented ginseng improved antibiotic‐induced diarrhoea and the gut microbiota profiles of rats

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Abstract

Aims

This study investigated the efficacy of Limosilactobacillus fermentum-fermented ginseng for improving colitis and the gut microbiota profiles in rats and explored the benefits of the L. fermentum fermentation process to ginseng.

Methods and results

Ginseng polysaccharide and ginsenoside from fermented ginseng were analysed by UV and HPLC. Antibiotic-fed rats were treated with fermented ginseng and a L. fermentum–ginseng mixture. Histopathology- and immune-related factors (TNF-α, IL-1β, IL-6 and IL-10) of the colon were assayed by using pathological sections and ELISA. After treatment, fermented ginseng relieved the symptoms of antibiotic-induced diarrhoea and colon inflammation, and the expression of colon immune factors returned to normal. The gut microbial communities were identified by 16S rRNA gene sequencing. The results showed that the alterations in the gut microbiota returned to normal. In addition, the gut microbiota changes were correlated with immune factor expression after treatment. The fermented ginseng had better biological functions than a L. fermentum–ginseng mixture.

Conclusions

Fermented ginseng can relieve diarrhoea and colon inflammation and restore the gut microbiota to its original state. The process of L. fermentum fermentation can expand the therapeutic use of ginseng.

Significance and impact of the study

This research suggested the potential function of fermented ginseng to relieve diarrhoea and recover the gut microbiota to a normal level and explored the benefits of the Limosilactobacillus fermentum fermentation process to ginseng.