Does the Wolf (Canis lupus) Exhibit Human Habituation Behaviours after Rehabilitation and Release into the Wild? A Case Report from Central Italy.

The knowledge of how wolves' movement patterns and habitat selection are affected by habituation to persons after a period of veterinary isolation, treatment and non-agonistic experience with humans is scarce. Unnatural behaviours could be transferred by imitation to members of the pack and to subsequent generations, increasing direct interaction risks. We used GPS data from a rescued radio-collared female wolf after an 11-day rehabilitation to estimate home range, movement patterns, circadian rhythms, and habitat selection, searching for signals of eventual behavioural distortions. In the period 1 August−26 November 2013, 870 valid locations were acquired. The wolf moved within a minimum convex polygon (95%) of 6541.1 ha (79% wooded), avoiding anthropized areas. Nocturnal and diurnal displacements were significantly different (p < 0.01). Nocturnal displacements were 4409.4 ± 617.5 m during summer and 3684.8 ± 468.1 m during autumn, without differences between seasons. Diurnal movements were significantly higher (p < 0.01) in the summer (2239.0 ± 329.0 m) than in the autumn (595.9 ± 110.3 m), when the hunting season was running. As for a wild wolf, clear complementarity concerning human activities was recorded and no habituation signals were detected, but this is only a first case study that aims to be a stimulus for further research and a call for widespread data sharing.

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.

The Effects of Diet Formulation on the Yield, Proximate Composition, and Fatty Acid Profile of the Black Soldier Fly (Hermetia illucens L.) Prepupae Intended for Animal Feed.

The black soldier fly (BSF; Hermetia illucens L.) is a very promising insect species due to the ability to convert low-value substrates in highly nutrient feed. This work aimed to study the effect of three nominally isoenergetic diets containing plant ingredients such as barley, alfalfa, and wheat byproducts, formulated to be higher in nonfiber carbohydrates (TMD1), fibers (TMD2), and protein (TMD3) in comparison to an extensively genetic modified cereal (cornmeal)-based diet (C), on the growth, yield, and nutritive traits of BSF prepupae (BSFPs). Three growing trials with four biological replicates were carried out. Proximate and fatty acid analyses were performed on the diets and BSFPs. Feed conversion ratios (FCR), dry matter and nutrient yields, and apparent concentration factors (aBCF) for fatty acids (FAs) were calculated. Diet formulation had a substantial effect on the survival, development rate, and larval yield, but the FCR was unaffected. The BSFPs fed TMD3 did not result in a higher crude protein content in comparison to the C or TMD2 diets. Despite the leveled fat content of the diets, BSFPs reared on TMD1 were highest in fat, saturated FA, and fat yield. An apparent bioconcentration factor (aBCF) value lower than unity that was found for the unsaturated FA suggests that the BSFPs inefficiently absorb them from the diet or possibly turn them into saturated FA. However, the unsaturated FA accumulation in BSFPs depended on the levels that were found in the diet, which suggested some possibilities for the FA profile modulation. Overall, the TMD2 performed well despite the low-value of its main ingredients and high fiber content and can be considered to be a feasible option for the mass rearing of BSFPs that are intended for animal feed.

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.

Evaluation of antibody response and nonspecific lymphocyte blastogenesis following inoculation of a live attenuated bluetongue virus vaccine in goats.

OBJECTIVE: To evaluate vaccine safety, antibody response, and nonspecific lymphocyte blastogenesis following inoculation of a commercial monovalent live attenuated bluetongue virus (BTV) serotype 2 vaccine in goats. ANIMALS: 12 nonpregnant and nonlactating Saanen goats. PROCEDURE: 6 goats were inoculated with the monovalent live attenuated BTV serotype 2 vaccine, which has been widely used in Italy during the proceding 2 years. The other 6 goats were unvaccinated and represented negative controls. Nonspecific lymphocyte blastogenesis was evaluated 14 and 7 days before and 7, 21, and 49 days after vaccination by measuring DNA synthesis in peripheral blood mononuclear cells (PBMCs) stimulated with phytohemagglutinin, concanavalin-A, and pokeweed mitogen. On the same days as lymphocyte blastogenesis, blood samples were taken to determine serum concentrations of anti-BTV antibodies. RESULTS: During the 7 weeks following vaccination, PBMCs obtained from vaccinated goats had a significantly decreased response to mitogens in terms of DNA synthesis, compared with PBMCs from the same goats before vaccination. Conversely during the experiment, no significant change was found in the response of the PBMCs obtained from unvaccinated goats. Starting from 21 days after vaccination, serum from vaccinated goats had anti-BTV antibodies. No anti-BTV antibodies were detected in the serum from unvaccinated goats. CONCLUSIONS AND CLINICAL RELEVANCE: Inoculation of goats with the monovalent live attenuated BTV serotype 2 vaccine described herein resulted in a profound depression of nonspecific lymphocyte blastogenesis, which might compromise the resistance of vaccinated goats to pathogens.

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.

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.

Heat shock modulates adipokines expression in 3T3-L1 adipocytes.

Studies have demonstrated that heat shock is associated with alteration in energy metabolism. In this study, we investigated the effect of heat shock on gene expression and secretion of adiponectin and leptin, and gene expression of Hspa2 and Ppargamma in 3T3-L1 adipocytes. Compared with 37 degrees C, adiponectin mRNA was higher at 39 degrees C, and lower at 41 degrees C. Leptin mRNA was higher when adipocytes were exposed to 41 degrees C compared with 37 and 39 degrees C. Secretion of adiponectin increased at 39 degrees C, and when cells were exposed to 41 degrees C it was not detectable. Leptin secretion increased significantly at 41 degrees C, compared with 37 and 39 degrees C. Hspa2 mRNA was increased at 39 degrees C, and the highest level was reached at 41 degrees C. Ppargamma mRNA exhibited a substantial increase in a temperature-dependent manner. The study provides the first evidence of a possible direct effect of heat shock on adiponectin and leptin gene expression and secretion, and demonstrates that the expression of the two adipokines is differentially regulated at the temperatures tested.

Lymphocyte functions in dairy cows in hot environment.

This study was carried out to ascertain the effects of intense high environmental temperatures (HET) on lymphocyte functions in periparturient dairy cows. The study was undertaken from the beginning of March through the end of July 2003 in a commercial dairy unit located approximately 40 km north of Rome. Thirty-four Holstein cows were utilised in the study. Twenty-two of these cows gave birth in spring (SP cows), from 28 March to 30 April. The remaining 12 cows gave birth in summer (SU cows), between 15 June and 2 July. The two groups of cows were balanced for parity and were fed the same rations. Blood samples were taken 4, 3, 2 and 1 week before calving, and 1, 2 and 4 weeks after calving, in order to evaluate peripheral blood mononuclear cell (PBMC) function in vitro, and to determine plasma cortisol concentrations. After isolation, the PBMC were stimulated with mitogens and their response in terms of DNA synthesis and IgM secretion was measured. During spring, either the day (9-20 h) or the night (21-8 h) temperature humidity index (THI) was below the upper critical THI (72) established for dairy cows. During summer, the mean daily THI values were 79.5+/-2.9 during the day and 70.1+/-4.7 during the night. Furthermore, during summer, three heat waves (a period of at least 3 consecutive days during which there were less than 10 recovery hours) occurred. Recovery hours were intended hours with a THI below 72. The first heat wave lasted 5 days, the second 6 days, and the third 15 days. Compared to the SP cows, over the entire periparturient period the extent of DNA synthesis and IgM secretion levels were lower (P ranging from <0.01 to 0.0001) and higher (P<0.01) respectively, in the SU cows. Before calving, the SU cows also presented higher (P<0.01) concentrations of plasma cortisol compared to the SP cows. This study indicates that the effects of HET on the immune response depend on the specific immune function under consideration, and that neuroendocrinal changes due to HET may play a role in the perturbation of immune functions.

Moderate summer heat stress does not modify immunological parameters of Holstein dairy cows.

The study was undertaken during spring and summer months in a territory representative of the Mediterranean climate to assess the effects of season on some immunological parameters of dairy cows. Twenty Holstein cows were used. Eleven of those cows gave birth during spring; the remaining nine cows gave birth in summer. The two groups of cows were homogeneous for parity. Values of air temperatures and relative humidity were recorded both during spring and summer, and were utilized to calculate the temperature humidity index (THI). One week before the expected calving, rectal temperatures and respiratory rates of the cows were recorded (1500 hours), and cell-mediated immunity was assessed by measuring the proliferation of mitogen-stimulated peripheral blood mononuclear cells (PBMC). Within 3 h of calving, one colostrum sample was taken from each cow and analysed to determine content of immunoglobulin (Ig) G1, IgG2, IgM and IgA. At 48 h after birth, passive immunization of the calves was assessed by measuring total serum IgG. During summer, daytime (0900-2000 hours) THI values were above the upper critical value of 72 [75.2, (SD 2.6)] indicating conditions that could represent moderate heat stress. That THI values were able to predict heat stress was confirmed by the values of rectal temperatures and respiratory rates, which were higher (P < 0.05 and P < 0.001 respectively) during summer. Proliferation of PBMC, the colostral concentration of Ig fractions and serum levels of IgG in their respective off-spring did not differ between spring and summer cows. Results indicated that moderate heat stress due to the hot Mediterranean summer does not modify cell-mediated immunity, the protective value of colostrum and passive immunization of the offspring in dairy cows.