Genomic responses to climatic challenges in beef cattle: A review.

Climate change is a major concern for the near future and for livestock breeding. Cattle breeding, due to its greenhouse gas emissions, is one of the most implicated industries. Consequently, the main future goals are to breed animals resilient to climate change, with the aim of lowering the livestock impact on the environment and selecting animals that will be able to resist different, unsuitable, and changing climates. The aim of this literature review is to compare the most recent studies on the response and adaptation of beef cattle breeds to extreme environments, in terms of genes and pathways involved. Beef breeding is just starting to implement genomics in its selection plans, and shedding light on the genomic responses to extreme climates could speed up and simplify the adaptation of these breeds to climate change. This review discusses the genes involved in climatic stress responses, including those related to extremely cold climates, in beef and dual-purpose cattle breeds. Genes were associated with productive traits, coat and skin structure and development, thermotolerance, cellular physiology and DNA repair mechanisms, immune system, and fertility traits. The knowledge of genes and pathways involved in climate resilience should be taken into consideration for further selection in beef cattle breeding and could promote the valorization of local breeds adapted to extreme environmental conditions. The use of local or resilient breeds could enhance the environmental and social sustainability, animal welfare, and production, compared with the introduction of cosmopolitan breeds with uncertain adaptation in uncontrolled environmental areas.

Carbon footprint of organic beef meat from farm to fork: a case study of short supply chain.

BACKGROUND: Sustainability of food systems is one of the big challenges facing humanity. Local food networks, especially those using organic methods, are proliferating worldwide, and little is known about their carbon footprints. This study aims to assess greenhouse gas (GHG) emissions associated with a local organic beef supply chain using a cradle-to-grave approach. RESULTS: The study determined an overall burden of 24.46 kg CO2  eq. kg-1 of cooked meat. The breeding and fattening phase was the principal source of CO2 in the production chain, accounting for 86% of the total emissions. Enteric methane emission was the greatest source of GHG arising directly from farming activities (47%). The consumption of meat at home was the second high point in GHG production in the chain (9%), with the cooking process being the main source within this stage (72%). Retail and slaughtering activities respectively accounted for 4.1% and 1.1% of GHG emissions for the whole supply chain. CONCLUSION: The identification of the major sources of GHG emissions associated with organic beef produced and consumed in a local food network may stimulate debate on environmental issues among those in the network and direct them toward processes, choices and habits that reduce carbon pollution. © 2018 Society of Chemical Industry.

Impact of Seasonal Conditions on Quality and Pathogens Content of Milk in Friesian Cows.

Heat stress negatively affects milk quality altering its nutritive value and cheese making properties. This study aimed at assessing the impact of seasonal microclimatic conditions on milk quality of Friesian cows. The study was carried out in a dairy farm from June 2013 to May 2014 at Beni-Suef province, Egypt. Inside the barn daily ambient temperature and relative humidity were recorded and used to calculate the daily maximum temperature-humidity index (mxTHI), which was used as indicator of the degree of heat stress. The study was carried out in three periods according to the temperature-humidity index (THI) recorded: from June 2013 to September 2013 (mxTHI>78), from October 2013 to November 2013 (mxTHI 72-78) and from December 2013 to April 2014 (mxTHI<72). Eighty Friesian lactating dairy cows were monitored in each period. The three groups of cows were balanced for days in milk and parity. Milk quality data referred to somatic cell count, total coliform count (TCC), faecal coliform count (FCC), Escherichia coli count, percentage of E. coli, and Staphylococcus aureus, percentage of fat, protein, lactose, total solid and solid non-fat. Increasing THI was associated with a significant decrease in all milk main components. An increase of TCC, FCC, and E. coli count from mxTHI<72 to mxTHI>78 was observed. In addition, the isolation rate of both S. aureus and E. coli increased when the mxTHI increased. The results of this study show the seriousness of the negative effects of hot conditions on milk composition and mammary gland pathogens. These facts warrant the importance of adopting mitigation strategies to alleviate negative consequences of heat stress in dairy cows and for limiting related economic losses.

Comparison of colostrum and milk extracellular vesicles small RNA cargo in water buffalo.

Recently, much interest has been raised for the characterization of signaling molecules carried by extracellular vesicles (EVs), which are particularly enriched in milk (mEVs). Such interest is linked to the capability of EVs to cross biological barriers, resist acidification in the gastric environment, and exert modulation of the immune system, mainly through their microRNA (miRNA) content. We characterized the small-RNA cargo of colostrum EVs (colosEVs) and mEVs from Italian Mediterranean buffalo through next generation sequencing. Colostrum (first milking after birth) and milk (day 50 of lactation) were sampled from seven subjects from five farms. ColosEVs and mEVs were subjected to morphological characterization, followed by high-depth sequencing of small RNA libraries produced from total RNA. The main difference was the amount of EV in the two samples, with colostrum showing 10 to 100-fold higher content than milk. For both matrices, miRNA was the most abundant RNA species (95% for colosEVs and 96% for mEVs) and three lists were identified: colosEV-specific, mEV-specific and shared most expressed. Gene ontology (GO) enrichment analysis on miRNA targets highlighted many terms related to the epigenetic, transcriptional and translational regulations across the three lists, with a higher number of enriched terms for colosEV-specific miRNAs. Terms specific to colosEVs were related to "cell differentiation" and "microvillus assembly", while for mEV "cardiac and blood vessel development" and "mitochondria" emergerd. Immune modulation terms were found for both sample-specific miRNAs. Overall, both matrices carry a similar molecular message in terms of biological processes potentially modulated into receiving cells, but there is significant difference in the abundance, with colostrum containing much more EVs than milk. Moreover, colosEVs carry molecules involved in signal transduction, cell cycle and immune response, as for mEVs and EVs of other previously characterized species, but with a special enrichment for miRNAs with epigenetic regulation capacities. These beneficial characteristics of colosEVs and mEVs are essential for the calf and could also be exploited for the therapeutic purposes in humans, although further studies are necessary to measure the sanitization treatment impact on EV conservation, especially in buffalo where milk is consumed almost exclusively after processing.

Effects of fatty acid mixtures on proliferation of peripheral blood mononuclear cells in dairy cows.

This in vitro study was performed to assess the effects of three different mixtures of nonesterifed fatty acids (NEFA) on mitogen-driven proliferation of peripheral blood mononuclear cells (PBMC) in dairy cows. Substantially, the three mixtures differed for n-6 to n-3 fatty acids (FA) ratio and were intended to mimic plasma NEFA composition of cows given fat supplements with different n-6 to n-3 FA ratio. PBMC from six Holstein heifers were cultured in media containing three different mixtures of oleic, palmitic, stearic, linoleic, palmitoleic, or linolenic acid at concentrations mimicking different degree of lipomobilisation. Proliferation of PBMC was stimulated by concanavalin A or pokeweed mitogen (PWM). Low concentrations of the three mixtures (62.5 and 125 µmol/l), did not affect the ability of PBMC to proliferate. Concentrations of the three mixtures mimicking medium-intense lipomobilisation (500, 1,000 and 1,500 µmol/l) impaired to the same extent proliferation of PBMC. The addition to cultures of the three mixtures at concentration of 250 µmol/l, impaired the proliferation only in PBMC stimulated with PWM. Even in this case, the three mixtures did not exert differential effects on PBMC proliferation. Present results reinforce the hypothesis that high concentrations of plasma NEFA play a role in the immunosuppression taking place in cows undergoing intense lipomobilisation, and authorize to suggest that under these conditions different composition of plasma NEFA in terms of different n-6 to n-3 FA ratio cannot prevent their negative effects on lymphocyte proliferation.

Responses of Dairy Buffalo to Heat Stress Conditions and Mitigation Strategies: A Review.

Increases in temperature and the greater incidence of extreme events are the consequences of the climate change that is taking place on planet Earth. High temperatures create severe discomfort to animal farms as they are unable to efficiently dissipate their body heat, and for this, they implement mechanisms to reduce the production of endogenous heat (reducing feed intake and production). In tropical and subtropical countries, where buffalo breeding is more widespread, there are strong negative consequences of heat stress (HS) on the production and quality of milk, reproduction, and health. The increase in ambient temperature is also affecting temperate countries in which buffalo farms are starting to highlight problems due to HS. To counteract HS, it is possible to improve buffalo thermotolerance by using a genetic approach, but even if it is essential, it is a long process. Two other mitigation approaches are nutritional strategies, such as the use of vitamins, minerals, and antioxidants and cooling strategies such as shade, fans, sprinklers, and pools. Among the cooling systems that have been evaluated, wallowing or a combination of fans and sprinklers, when wallowing is not available, are good strategies, even if wallowing was the best because it improved the production and reproduction performance and the level of general well-being of the animals.

In vitro antioxidant and anti-inflammatory activity of an oleuropein-enriched extract obtained from olives leaves on BME-UV1 cells.

In this in vitro study, for the first time was evaluated the antioxidant and anti-inflammatory effect of an Oleuropein-enriched extract (OleE) on bovine mammary epithelial cell line (BME-UV1). OleE was obtained from olives leaves and characterized by HPLC and NMR analysis. Cell viability test indicated that OleE at concentrations of 7.8 up to 250 µg/mL did not exert cytotoxic effect. At concentration of 31.2 up to 250 µg/mL, a dose dependent reduction of ROS production induced by hydrogen peroxide was observed. In addition, OleE at 62.5, 125 and 250 µg/mL showed a dose-dependent reduction in gene expression of TNF, IL1B, and IL10 after exposure to LPS. The downregulation of ROS production and cytokines expression in BME-UV1 by OleE confirmed the antioxidant and anti-inflammatory properties. In vivo experiments will be necessary for future applications of OleE as natural feed supplement in dairy cattle to reduce incidence of oxidative stress in peripartal period.

Effect of Antioxidant Supplementation on Milk Yield and Quality in Italian Mediterranean Lactating Buffaloes.

Buffaloes are raised mainly to obtain milk that is nutritionally very rich. The technological characteristics of buffalo milk are optimal for processing into cheese, and it is mainly used to produce mozzarella cheese. Under stressful conditions, buffaloes, like other animals, produce milk qualitatively poorly. The stressors that can affect the quality of production are, in addition to other factors, deficiencies in nutrients such as vitamins, antioxidants, and minerals. In this study, we evaluated the effect of antioxidant supplementation on the quality of buffalo milk. Sixty-six buffaloes were enrolled and subdivided into two balanced groups of 33 each. The ZnSe group received 0.2 kg/head/day of Bufalo Plus® containing antioxidants and barley meal, CaCO3 and MgCO3 mix; the control group was supplemented with 0.2 kg/head/day of barley meal, CaCO3 and MgCO3 mix. The two groups were fed ad libitum with a total mixed ration (TMR). The amount of diet distributed was recorded daily, and the residue in the trough manger was recorded three times per week. TMR samples were taken every two weeks for each group. Daily milk yield was recorded twice a week. Milk samples were collected every four weeks and analysed for chemical and technological properties. Furthermore, milk total antioxidant capacity was determined. The results obtained showed that the antioxidant supplement had no effect on feed intake, feeding behaviour, and feed efficiency. The treatment positively influenced milk production while it did not affect the chemical characteristics of the milk. In addition, the supplement of antioxidants improved the milk clotting properties (MCP). The supplement did not affect the antioxidant activity of the milk.

Dynamics of the temperature-humidity index in the Mediterranean basin.

The study was aimed at describing the temperature humidity index (THI) dynamics over the Mediterranean basin for the period 1951-2007. The THI combines temperature and humidity into a single value, and may help to predict the effects of environmental warmth in farm animals. In particular, on the basis of THI values, numerous studies have been performed to establish thresholds for heat stress in dairy cows. The THI was calculated by using monthly mean values of temperature and humidity obtained from the National Center for Environmental Prediction/National Center for Atmospheric Research reanalysis project. The analysis demonstrated a high degree of heterogeneity of THI patterns over the Mediterranean basin, a strong north-south gradient, and an overall warming during the study period, which was particularly marked during summer seasons. Results indicated that several areas of the basin present summer THI values which were unfavorable to cow welfare and productivity, and that risk of heat stress for cows is generally greater in the countries of the south coast of the basin. Furthermore, THI data from the summer 2003 revealed that severe positive anomalies may impact areas normally characterized by a favorable climate for animal production. In conclusion, THI dynamics should be taken into careful consideration by farmers and policy makers operating in Mediterranean countries when planning investments in the sector of animal production. The investments should at least partially be directed towards implementation of adaptation measures, which may help to alleviate the impact of hot on farm animals welfare, performance and health.

Carbon Footprint of Mediterranean Pasture-Based Native Beef: Effects of Agronomic Practices and Pasture Management under Different Climate Change Scenarios.

A better understanding of soil organic carbon (SOC) dynamics is needed when assessing the carbon footprint (CFP) of livestock products and the effectiveness of possible agriculture mitigation strategies. This study aimed (i) to perform a cradle-to-gate CFP of pasture-based beef cattle in a Mediterranean agropastoral system (ii) and to assess the effects on the CFP of alternative tillage, fertilizing, and grazing practices under current (NCC) and future climate change (CC) scenarios. Minimum (Mt) and no-tillage (Nt) practices were compared to current tillage (Ct); a 50% increase (Hf) and decrease (Lf) in fertilization was evaluated against the current (Cf) rate; and rotational grazing (Rg) was evaluated versus the current continuous grazing (Cg) system. The denitrification-decomposition (DNDC) model was run using NCC as well as representative concentration pathways to investigate the effects of farm management practices coupled with future CC scenarios on SOC dynamics, N2O fluxes, and crop yield. Within NCC and CtCf, an emission intensity of 26.9 ± 0.7 kg CO2eq per kg live body weight was estimated. Compared to Ct, the adoption of Mt and Nt reduced the CFP by 20% and 35%, respectively, while NtHf reduced it by 40%. Conservation tillage practices were thus shown to be effective in mitigating greenhouse gas emissions.

Antioxidant and anti-inflammatory effects of pomegranate peel extracts on bovine mammary epithelial cells BME-UV1.

Pomegranate peel extracts (PPE) were tested for the first time on BME-UV1, a valid cellular model to study the bovine mammary epithelial metabolism, to evaluate the effects on the oxidative stress and inflammatory status. Based on the statistical analysis of MTT data, PPE at 0.1, 1.0 and 10 µg/mL resulted not cytotoxic after 24 h, 48 h and 7 days of treatment. At the same concentrations, PPE induced a reduction of ROS production elicited by the addition of hydrogen peroxide or lipopolysaccharide evidencing an antioxidant effect confirmed also by a decrease of malondialdehyde. At 10 µg/mL, PPE reduced pro-inflammatory cytokines expressions showing an anti-inflammatory effect on BME-UV1 treated with lipopolysaccharide. Although in vivo experiments are necessary, the results of this study are promising for future applications of PPE as feed supplement for dairy cattle, in particular around calving, when the animals are more subject to oxidative stress and inflammatory diseases.

Sowing Date and Seeding Rate Affect Bioactive Compound Contents of Chickpea Grains.

Chickpea grains may represent an alternative to soybean meals and energetic concentrates in animal feeding, as their nutritional value can help to increase the sustainability of livestock systems. Unfortunately, the presence of bioactive compounds with anti-nutritional effects can prevent its direct use, especially in mono-gastrics. It is known that the synthesis of these compounds depends on genetic expression, which is also influenced by growth conditions. The objective of this two-year study was to assess the effect of sowing date (winter versus spring) and seeding rate (70 versus 110 seeds m-2) on the accumulation of soluble carbohydrates, α-galactosides, trypsin inhibitors, and inositol phosphates in the grains of two Kabuli cultivars, in the Mediterranean climate. The results showed that seeds collected from winter sowing contained more trypsin inhibitors than those seeded in spring (+ 4%, on average), reaching values between 16.1 and 18.6 TIU mg protein-1. The seeding rate affects only the α-galactosides content, which increases (+9%) at lower densities (70 seeds m-2). These findings suggest that agronomic management can be used to modulate the content of some anti-nutritional factors in the seeds, even though the genetic characteristics and phenotypic expression, in relation to the climatic conditions, seem to deeply affect the content of all the bioactive compounds investigated.

Surveying selected European feed and livestock production chains for features enabling the case-specific post-market monitoring of livestock for intake and potential health impacts of animal feeds derived from genetically modified crops.

This review, which has been prepared within the frame of the European Union (EU)-funded project MARLON, surveys the organisation and characteristics of specific livestock and feed production chains (conventional, organic, GM-free) within the EU, with an emphasis on controls, regulations, traceability, and common production practices. Furthermore, an overview of the origin of animal feed used in the EU as well as an examination of the use of genetically modified organisms (GMOs) in feed is provided. From the data, it shows that livestock is traceable at the herd or individual level, depending on the species. Husbandry practices can vary widely according to geography and animal species, whilst controls and checks are in place for notifiable diseases and general health symptoms (such as mortality, disease, productive performance). For feeds, it would be possible only to make coarse estimates, at best, for the amount of GM feed ingredients that an animal is exposed to. Labeling requirements are apparently correctly followed. Provided that confounding factors are taken into account, practices such as organic agriculture that explicitly involve the use of non-GM feeds could be used for comparison to those involving the use of GM feed.

Effects on functions of ovine blood mononuclear cells for each of several fatty acids at concentrations found in plasma of healthy and ketotic ewes.

OBJECTIVE: To assess effects on functions of peripheral blood mononuclear cells (PBMC) obtained from ewes for each of several fatty acids represented in ovine plasma at concentrations mimicking those of ketotic or healthy ewes. SAMPLE POPULATION: Blood samples obtained from 6 Sardinian ewes. PROCEDURE: The PBMC were cultured in media that contained oleic (OA), palmitic (PA), stearic (SA), linoleic (LA), or palmitoleic (POA) acid at concentrations similar to those of ketotic or healthy ewes. Synthesis of DNA was stimulated by use of concanavalin A or pokeweed mitogen (PWM). Secretion of IgM was stimulated by use of PWM. RESULTS: High concentrations (900, 450, and 225 micromol/L) of OA significantly inhibited DNA synthesis and IgM secretion of PBMC. Conversely, low concentrations (56 or 28 micromol/L) of OA significantly enhanced DNA synthesis of PBMC. High concentrations of PA (600, 300, 150, 75, 375, or 18.7 micromol/L) and SA (300, 150, or 75 micromol/L) significantly inhibited DNA synthesis of PBMC. High concentrations of PA (600, 300, 150, 75, 375, or 18.7 micromol/L) and SA (300, 150, 75, or 38 micromol/L) also significantly inhibited IgM secretion of PBMC. None of the concentrations of LA and POA affected PBMC functions. CONCLUSION AND CLINICAL RELEVANCE: Impaired immunoresponsiveness of ketotic ewes is likely associated with an increase of plasma concentrations of OA, PA, or SA and not with that of LA or POA. At physiologic concentrations, single fatty acids are likely to participate in modulation of immunoresponsiveness by exerting suppressive or stimulatory effects on immune cells.

Effects of nonesterified fatty acids and beta-hydroxybutyrate on functions of mononuclear cells obtained from ewes.

OBJECTIVE: To assess the effects of nonesterified fatty acids (NEFA) and beta-hydroxybutyrate (BHBA) on functions of mononuclear cells obtained from ewes. ANIMALS: 6 Sardinian ewes. PROCEDURE: Mononuclear cells were cultured with concentrations of NEFA (0, 15.6, 31.2, 62.5, 125, 250, 500, 1,000, or 2,000 micromol/L) and BHBA (0, 0.45, 0.9, 1.8, or 3.6 mmol/L). Concentrations of NEFA and BHBA were intended to mimic those of ketotic or healthy ewes, and NEFA and BHBA were tested alone and in combination. Synthesis of DNA was stimulated by use of concanavalin A (Con A) or pokeweed-mitogen (PWM). Secretion of IgM was stimulated by use of PWM. RESULTS: Synthesis of DNA stimulated by Con A and PWM was significantly inhibited by high concentrations of NEFA (> or = 250 micromol/L) or by a combination of high concentrations of NEFA (> or = 250 micromol/L) and all concentrations of BHBA (> or = 0.45 mmol/L). In contrast, DNA synthesis was not inhibited by low concentrations of NEFA (< or = 125 micromol/L) or by a combination of low concentrations of NEFA (< or = 125 micromol/L) and the lowest concentration of BHBA (0.45 mmol/L). Secretion of IgM was significantly inhibited by all concentrations of NEFA and by all combinations of NEFA and BHBA concentrations. When used alone, none of the concentrations of BHBA inhibited DNA synthesis or IgM secretion. CONCLUSIONS AND CLINICAL RELEVANCE: Reduced immunoresponsiveness during ketosis is likely to be associated with an increase in plasma concentration of NEFA and not with an increase in plasma concentration of BH BA.

Chronic heat stress up-regulates leptin and adiponectin secretion and expression and improves leptin, adiponectin and insulin sensitivity in mice.

Heat stress (HS) induces adaptive responses that are responsible for alterations of carbohydrate and lipid metabolism. This study aimed to evaluate the effects of chronic heat treatment on the expression and secretion of leptin and adiponectin, important regulators of energy homeostasis, food intake and insulin action. C57BL/6 mice were subdivided into three groups (24 mice each). The first group was kept under control conditions (C: 22±2 °C). The second group was exposed to HS (35±1 °C). The third group was kept under control conditions and was food restricted (FR). The HS group had higher rectal temperature than the C and FR groups and lower food intake than the C group. Hspa1 (Hspa1a) gene expression in adipose tissue, muscle and liver was higher under HS than FR and C. Heat treatment resulted in decreased blood glucose and non-esterified fatty acids; increased leptin, adiponectin and insulin secretion; and greater glucose disposal. Leptin, adiponectin, leptin and adiponectin receptors, insulin receptor substrate-1 and glucose transporter mRNAs were up-regulated in HS mice. This study provides evidence that HS improves leptin and adiponectin signalling in adipose tissue, muscle and liver. Heat stress was responsible for improving insulin sensitivity and glucose uptake in peripheral tissues, probably mediated by adipokines. Changes in the adipokine levels and sensitivity to them may be considered as an adaptive response to heat.

Cellular thermotolerance is associated with heat shock protein 70.1 genetic polymorphisms in Holstein lactating cows.

Heat shock proteins (Hsp) are known to protect cells from several stressors. Nucleotide changes in the flanking regions [5'- and 3'-untranslated region (UTR)] of Hsp gene might affect inducibility, degree of expression, or stability of Hsp70 mRNA. The present study aimed to investigate the association between inducible Hsp70.1 single nucleotide polymorphisms (SNPs) and heat shock (HS) response of peripheral blood mononuclear cells (PBMC) in dairy cows. Four hundred forty-six Italian Holstein cows were genotyped for four Hsp70.1 SNPs: g895 C/- and g1128 G/T in 5'-UTR, and g2154 G/A and g64 G/T in 3'-UTR. Genetic polymorphisms in 3'-UTR of bovine Hsp70.1 gene resulted monomorphic. Distribution of alleles of the nucleotide sequence polymorphism within the 5'-UTR of the bovine Hsp70.1 gene were 81.2% and 18.8% for C and -, respectively, and 77.8% and 22.2% for G and T, respectively. Among the 446 genotyped animals, a group of cows balanced for days in milk and parity was selected to be representative of the following genotypes: CC (n = 8), C- (n = 7), and -- (n = 7) and GG (n = 8), GT (n = 11), and TT (n = 3) in 5'-UTR. PBMC were isolated from blood samples and heated at 43°C in thermal bath for 1 h and then incubated at 39°C in atmosphere of 5% CO(2) for 1, 2, 4, 8, 16, and 24 h (recovery times). Cell viability was determined by XTT assay. Gene and protein expression of Hsp70.1 was determined by real-time reverse transcription-polymerase chain reaction and by ELISA assay, respectively. For the two SNPs detected, one allele was the most frequent (C, 66.8% and G, 56.8%). Genotypes -- and TG showed higher (P < 0.05) viability compared with CC and GG, respectively. Genotypes C- and TT had intermediate viability. Gene expression of Hsp70.1 showed higher (P < 0.001) levels in -- and TG genotype compared with their counterparts. Genotypes -- and TG showed the higher level of inducible Hsp70.1 protein in respect to C-, TT and CC, GG. In conclusion, exposure to HS differently affected cell viability and gene and protein expression of Hsp70.1 in the selected genotypes. These results indicate that the presence of SNPs (C/- and G/T) in the 5'-UTR region of inducible Hsp70.1 ameliorates HS response and tolerance to heat of bovine PBMC. These mutation sites may be useful as molecular genetic markers to assist selection for heat tolerance.

Aflatoxin B1 and fumonisin B1 affect the oxidative status of bovine peripheral blood mononuclear cells.

Mycotoxins are secondary metabolites having a high cytotoxic potential. They are produced by molds and released in food and feed. To date, the mechanisms underlying the mycotoxin-induced cytotoxicity have not been fully clarified. The induction of oxidative stress, as a possible mechanism, has been postulated. This in vitro study was focused on the effect of two widely occurring mycotoxins, aflatoxin B(1) (AFB(1)) and fumonisin B(1) (FB(1)), on the oxidative status of bovine peripheral blood mononuclear cells (PBMC) incubated for 2 and 7 days at different levels of AFB(1) (0, 5 and 20 µg/ml) and FB(1) (0, 35 and 70 µg/ml). Reactive oxygen metabolites (ROM), intracellular thiols (SH), malondialdehyde (MDA) and gene expression of cytoplasmic superoxide dismutase (SOD) and glutathione peroxidase (GSHPX-1) were measured on PBMC after incubation. The highest concentration of AFB(1) and all concentrations of FB(1) caused an increase (p<0.05) of intracellular ROM without any time dependent effect. Intracellular SH decreased with 20 µg AFB(1)/ml (p<0.05) and the effect was particularly marked after 7 days of exposure. Intracellular SH were not affected by FB(1) even though a lower (p<0.05) SH level after 2 days exposure than after 7 days was observed. MDA increased (p<0.05) in AFB(1) or FB(1) treated PBMC. The exposure to FB(1) for 7 days increased MDA (p<0.05) only in cells treated with 70 µg/ml. Exposure of PBMC to AFB(1) reduced SOD mRNA while FB(1) decreased both SOD and GSHPX-1 mRNA abundance. These results demonstrate that, even though by different mechanisms, AFB(1) and FB(1) may induce cytotoxicity through an impairment of the oxidative status of PBMC.

The Hsp72 response in peri-parturient dairy cows: relationships with metabolic and immunological parameters.

The study was aimed at assessing whether the peri-parturient period is associated with changes of intracellular and plasma inducible heat shock proteins (Hsp) 72 kDa molecular weight in dairy cows, and to establish possible relationships between Hsp72, metabolic, and immunological parameters subjected to changes around calving. The study was carried out on 35 healthy peri-parturient Holstein cows. Three, two, and one week before the expected calving, and 1, 2, 3, 4, and 5 weeks after calving, body conditions score (BCS) was measured and blood samples were collected to separate plasma and peripheral blood mononuclear cells (PBMC). Concentrations of Hsp72 in PBMC and plasma increased sharply after calving. In the post-calving period, BCS and plasma glucose declined, whereas plasma nonesterified fatty acids (NEFA) and tumor necrosis factor-alpha increased. The proliferative responses of PBMC to lipopolysaccharide (LPS) declined progressively after calving. The percentage of PBMC expressing CD14 receptors and Toll-like receptors (TLR)-4 increased and decreased in the early postpartum period, respectively. Correlation analysis revealed significant positive relationships between Hsp72 and NEFA, and between PBMC proliferation in response to LPS and the percentage of PBMC expressing TLR-4. Conversely, significant negative relationships were found between LPS-triggered proliferation of PBMC and both intracellular and plasma Hsp72. Literature data and changes of metabolic and immunological parameters reported herein authorize a few interpretative hypotheses and encourage further studies aimed at assessing possible cause and effect relationships between changes of PBMC and circulating Hsp72, metabolic, and immune parameters in dairy cows.