A matter of age? How age affects the adaptation of lactating dairy cows to virtual fencing.

Virtual Fencing (VF) can be a helpful technology in managing herds in pasture-based systems. In VF systems, animals wear a VF collar using global positioning, and physical boundaries are replaced by virtual ones. The Nofence (Nofence AS, Batnfjordsøra, Norway) collars used in this study emit an acoustic warning when an animal approaches the virtual boundaries, followed by an aversive electrical pulse if the animal does not return to the defined area. The stimuli sequence is repeated up to three times if the animal continues to walk forward. Although it has been demonstrated that animals successfully learn to adapt to the system, it is unknown if this adaptation changes with animal age and thus has consequences for VF training and animal welfare. This study compared the ability of younger and older dairy cows to adapt to a VF system and whether age affected activity behavior, milk yield, and animal long-term stress under VF management. The study was conducted on four comparable strip-grazing paddocks. Twenty lactating Holstein-Friesian cows, divided into four groups of five animals each, were equipped with VF collars and pedometers. Groups differed in age: two groups of older cows (>4 lactations) and two groups of younger ones (first lactation). After a 7-d training, paddock sizes were increased by successively moving the virtual fence during four consecutive grazing periods. Throughout the study, the pedometers recorded daily step count, time spent standing, and time spent lying. For the determination of long-term stress, hair samples were collected on the first and last day of the trial and the hair cortisol content was assessed. Data were analyzed by generalized mixed-effect models. Overall, age had no significant impact on animal responses to VF, but there were interaction effects of time: the number of acoustic warnings in the last period was higher in younger cows (P < 0.001), and the duration of acoustic warnings at training was shorter for older cows (P < 0.01). Moreover, younger cows walked more per day during the training (P < 0.01). Finally, no effects on milk yield or hair cortisol content were detected. In conclusion, all cows, regardless of age, adapted rapidly to the VF system without compromising their welfare according to the indicators measured.

For dairy farmers, pasture management is a difficult task, including feeding the herd on demand, improving pasture use efficiency, and dealing with high labor costs. Virtual Fencing (VF) is an innovative technology that can help farmers to solve these issues. In a VF system animals wear a tracking collar. Physical boundaries are replaced by virtual ones using a smartphone application. The collars emit an acoustic warning when the animal reaches the virtual boundaries, further accompanied by an aversive electrical pulse if the animal does not return to the predefined area. Previous studies have shown that cattle learned to adapt to the system easily, but it is still unclear if older animals can adapt just as quickly. Thus, this is the first study investigating the effect of dairy cow age on learning VF in a strip-grazing trial. The results showed that older and younger cows adapted to the system equally fast, with no differences in activity behavior or changes in daily milk yield. Moreover, hair cortisol levels did not indicate lasting stress in the cows associated with the VF management during the trial. These results demonstrate the potential of VF in the management of lactating grazing cows of all ages.

Assessing the effectiveness of cannabidiol additive supplementation on canine behavior and cortisol levels.

In veterinary medicine, Cannabis has been used to treat pain conditions, inflammation, and seizures. However, little is known about its effect on dogs' behavior. This preliminary research aims to address this knowledge gap by evaluating the effectiveness of cannabidiol (CBD) oil in canine behavioral therapy. Twenty dogs, diagnosed with behavioral disorders and housed in a municipal shelter, participated in a double-blind trial. Ten dogs received CBD oil treatment, while the other ten received a control oil without CBD. Before (T0) and after (T1) the treatment, all the dogs underwent a temperament test to assess their behavior in the presence of four different stimuli: a human stranger, a novel object, a child-like doll, and a conspecific (another dog). Each stimulus was presented individually, and the dogs' behaviors were recorded on video and analyzed. Additionally, hair samples were collected using a shave-reshave technique for cortisol determination through Radio-Immuno-Assay. No behavioral differences were found between the two groups at both T0 and T1. There were no significant differences in the behavioral responses of either group when comparing T0 and T1. However, individual responses to the CBD oil treatment appeared to vary among subjects. A significant increase in hair cortisol levels (p-value <0.05) was observed in the group treated with CBD oil [T0 = 1.60 (1.44-1.93) pg/mg, T1 = 4.81(2.57-6.01) pg/mg]. These findings highlight the importance of individualized treatment when using Cannabis and encourage further research on the use of CBD oil in animal behavioral medicine.

Limitations of Spatial Judgment Bias Test Application in Horses (Equus ferus caballus).

Affective states are of increasing interest in the assessment of animal welfare. This research aimed to evaluate the possible limitations in the application of a spatial judgment bias test (JBT) in horses, considering the influence of stress level, personality traits, and the possible bias due to the test structure itself. The distinction between two positions, one rewarded (Positive) and the other not (Negative), was learned by 10 horses and 4 ponies,. Then, the latency to reach three unrewarded ambiguous positions (Near Positive, Middle, Near Negative) was measured. Furthermore, the validated Equine Behavior Assessment and Research Questionnaire (E-BARQ) was employed to assess personality traits. Fecal and hair cortisol levels were measured through radioimmunoassay (RIA), and the frequency of behavioral stress indicators was recorded. Results showed that horses that had the rewarded position (Positive) on the right approached Near Negative and Middle faster than those that had Positive on the left. Certain personality traits influenced the latency to reach Middle and Near Positive, but chronic stress did not seem to affect horses' judgment bias. This preliminary study highlighted several limitations in the employment of spatial JBT for the assessment of affective state in horses and that personality traits can partially influence the cognitive process. Further research is needed to refine the use of this test in horses, considering the peculiarities both of species and of individuals.

Anogenital scent-marking signals fertility in a captive female Alaotran gentle lemur.

The Lake Alaotra gentle lemur (Hapalemur alaotrensis) is one of the 25 most endangered primates in the world and shows low success rate in captive breeding programmes. It is therefore vital to further understand its reproductive biology. We studied a captive troop consisting of five individuals hosted at Jersey Zoo during breeding and non-breeding periods over 1 year. We collected behavioural data (n = 318 h) using all occurrence of some behaviours and ad libitum sampling methods, as well as faecal (n = 54) and anogenital scent (n = 35) samples of the breeding female. We measured sex hormone levels using enzyme immunoassay technique and investigated the volatile component of odour signals using solid-phase microextraction and gas chromatography-mass spectrometry. We observed sexual and aggressive behaviours occasionally during the breeding period. Our regression analysis showed that only period significantly predicted rates of female anogenital scent-marking, whereby the female performed anogenital scent-marking more frequently during the breeding rather than the non-breeding period. In contrast, female hormone levels did not significantly explain variation in rates of neither male nor female olfactory, sexual and affiliative behaviours, suggesting that individuals' behaviour alone is not an effective indicator of the ovulation window. The volatile chemical profile of anogenital odour secretions changed over the study, with four compounds distinguishing the fertile window during the breeding period. In conclusion, our findings suggest that anogenital scent-marking may signal the reproductive status of captive female gentle lemurs.

Assessment of blood and productive parameters in mid-lactation dairy cows fed different diets: replacement of corn silage with triticale silage.

Corn crops require large amounts of resources that affect the environmental sustainability of dairy cow farming systems. The aim of the study was thus to investigate the effects of the replacement of corn silage (CS) with triticale silage (TS) by evaluating blood and productive parameters. The study lasted 7 weeks and involved two groups of 20 Italian Holstein Friesian dairy cows that were homogeneous in terms of parity ( 3 ± 1.5 ), days in milk (DIM) ( 150 ± 85.0 ), and daily milk production ( 26 ± 4.6  kg). Chemical analysis of feeds was carried out weekly. Dry-matter intake was estimated daily. At the beginning and end of the trial, haematological, metabolic, and immunological parameters were analysed. At the same, time body weight and body condition score were measured. Milk characteristics were also analysed weekly. Statistical analysis was performed by ANOVA on data of the second sampling, and a non-parametric test was performed to analyse BCS. Regarding the haematological parameters in the two groups, only lymphocyte values were not in the normal range (2.86 and 2.50 × 10 9  L for CS and TS, respectively). Metabolic parameters were in the normal range except for blood ureic nitrogen (BUN; 13.65 and 14.04  mg dL - 1 ), non-esterified fatty acids (NEFAs; 21.40 and 31.93  µ mol L - 1 ), and Cl (91.99 and 93.50  mmol L - 1 ). Hair cortisol was low (0.94 and 0.91  pg mg - 1 ), indicating the absence of stress signs, as confirmed by the results of other immunological parameters (serum lysozyme (SL), bactericidal activity (SBA), haptoglobin (HP), and oxygen free radicals (OFRs)). Statistical differences were not found either for haematological or biochemical parameters. The total replacement of CS with TS did not affect milk yield and composition. In conclusion, the replacement of CS by TS did not give rise to significant modifications in the parameters investigated and did not alter the health status of the animals, thus suggesting the feasibility of its introduction into the diet of mid-lactation dairy cows.

A field-friendly alternative to freeze-drying faeces for glucocorticoid metabolite analyses of African wild dogs (Lycaon pictus).

Endocrine studies using faeces as hormone matrix have become increasingly popular to examine adrenocortical activity in wildlife. A prerequisite for this approach is to minimize alteration of faecal glucocorticoid metabolite (fGCM) composition post-defecation. This is done by freezing the collected material as soon as possible after collection, and removing moisture from the frozen faecal samples afterwards (usually by freeze-drying). In remote areas, freeze-drying opportunities are often limited, and in the case of the African wild dog (Lycaon pictus), established assays revealed that fGCM concentrations remain comparable for only ∼24h post-defaecation. In the present study, three cost-effective drying treatments (exposure to sunlight, placement in a solar oven, and use of a food dehydrator) were investigated as alternatives to the golden standard of freeze-drying faeces.•In comparison to freeze-dried material, African wild dog faecal samples dried through sunlight exposure, a solar oven, and use of a food dehydrator revealed no significant differences in respective fGCM concentrations measured.•A food dehydrator would be the preferable option to dry African wild dog faeces if limited electrical supply is available. This technique dries faeces the fastest, and negates any reliance on weather conditions.

Evaluation of an enrichment programme for a colony of long-tailed macaques (Macaca fascicularis) in a rescue centre.

Long-tailed macaques are highly social primates that are commonly used in biomedical research as animal models. The aim of this study was to evaluate the effects of different kinds of enrichment on the behaviour and faecal cortisol metabolite (FCM) level in a colony of ex-laboratory long-tailed macaques during a programme of rehabilitation. The research was carried out in three periods, divided into two sessions each. Every period was composed of one control session (SC) and one session characterised by one type of enrichment: feeding enrichment (FE), manipulative enrichment (ME), and the last session during which manipulative and feeding enrichment were provided every day but in a mixed way (MIX). The results showed that manipulative and mixed enrichments caused positive changes to the activity budget of the colony, with a decrease in abnormal behaviour rates and an increase in play compared with control sessions. The rate of affiliative behaviours and low rate of aggression were probably because the group was composed mostly of females and it was stable, with a well-defined hierarchy. The research underlines the importance of a well-studied enrichment programme for the welfare of captive animals, which should exploit species-specific motivations.

Effects of scent enrichment on behavioral and physiological indicators of stress in zoo primates.

Captive breeding is vital for primate conservation, with modern zoos serving a crucial role in breeding populations of threatened species and educating the general public. However, captive populations can experience welfare issues that may also undermine their reproductive success. To enhance the wellbeing of endangered zoo primates, we conducted a study to assess the effects of a new scent enrichment program on captive red-ruffed lemurs (Varecia rubra), black howler monkeys (Alouatta caraya), siamangs (Symphalangus syndactylus), lar gibbons (Hylobates lar) and orangutans (Pongo pygmaeus pygmaeus). We combined behavioral observations and fecal endocrinology analyses to evaluate the effects of a series of essential oils (benzoin, lavender, lemongrass) on five captive troops (N = 19) housed at Dudley Zoo & Castle and Twycross Zoo (UK). We recorded observations of natural species-specific and abnormal stress-related behaviors for 480 h using instantaneous scan sampling. We collected 189 fecal samples and measured the fecal cortisol concentrations using radioimmunoassay. We found a significant effect of the scent enrichment on behaviors, with red-ruffed lemurs and black howler monkeys reducing their social interactions, as well as red-ruffed lemurs and lar gibbons decreasing their stress-related behaviors after they were exposed to the series of essential oils. We also found that red-ruffed lemurs displayed a significant increase in fecal glucocorticoids following exposure to essential oils. Our contradictory findings suggest that the effects of this series of essential oils may change depending on the species-specific social lives and olfactory repertoires of primates. In conclusion, we cannot recommend using these essential oils widely with zoo primates without additional evaluation.

Evaluation of Horses' Daytime Activity Budget in a Model of Ethological Stable: A Case Study in Italy.

The increasing interest in animal welfare and the knowledge of equine physiological and ethological needs have led to the development of different types of horses' management and housing systems. The research presented here aimed to assess the daytime activity budget of horses. Focal animal sampling was used as an observational sampling method, and the five animals were observed for a total of 9920 minutes in the paddock and inside the stall. The results showed that horses spent most of the daytime in foraging behaviors, followed by resting behaviors, and locomotion. Social behaviors (s.e. allogrooming, olfactory investigation) were rare, and the stereotypic behaviors (s.e. oral and locomotor stereotypies) occupied 2.74%±2.74% of the total time. The percentage of time spent in foraging, resting, and locomotion, reflects the activity budget observed in free-roaming feral horses. However, the rare occurrence of positive social interactions and the presence of some stereotypies could be aspects to ameliorate. This kind of housing facility could be considered a good alternative to traditional management; indeed, it might offer a better trade-off between the needs of the horse and the management goals from humans.

Non-invasive Assessment of Fecal Stress Biomarkers in Hunting Dogs During Exercise and at Rest.

Intense exercise causes to organisms to have oxidative stress and inflammation at the gastrointestinal (GI) level. The reduction in intestinal blood flow and the exercise-linked thermal damage to the intestinal mucosa can cause intestinal barrier disruption, followed by an inflammatory response. Furthermore, the adaptation to exercise may affect the gut microbiota and the metabolome of the biofluids. The aim of the present research was to evaluate the presence of a GI derangement in hunting dogs through a non-invasive sampling as a consequence of a period of intense exercise in comparison with samples collected at rest. The study included nine dogs that underwent the same training regime for hunting wild boar. In order to counterbalance physiological variations, multiple-day replicates were collected and pooled at each experimental point for each dog. The samples were collected immediately at rest before the training (T0), after 60 days of training (T1), after 60 days of hunting wild boar (T2), and finally, at 60 days of rest after hunting (T3). A number of potential stress markers were evaluated: fecal cortisol metabolites (FCMs) as a major indicator of altered physiological states, immunoglobulin A (IgA) as an indicator of intestinal immune protection, and total antioxidant activity [total antioxidant capacity (TAC)]. Since stool samples contain exfoliated cells, we investigated also the presence of some transcripts involved in GI permeability [occludin (OCLN), protease-activated receptor-2 (PAR-2)] and in the inflammatory mechanism [interleukin (IL)-8, IL-6, IL-1b, tumor necrosis factor alpha (TNFα), calprotectin (CALP), heme oxygenase-1 (HO-1)]. Finally, the metabolome and the microbiota profiles were analyzed. No variation in FCM and IgA content and no differences in OCLN and CALP gene expression between rest and training were observed. On the contrary, an increase in PAR-2 and HO-1 transcripts, a reduction in total antioxidant activity, and a different profile of microbiota and metabolomics data were observed. Collectively, the data in the present study indicated that physical exercise in our model could be considered a mild stressor stimulus.

Natural Horse Boarding Vs Traditional Stable: A Comparison of Hormonal, Hematological and Immunological Parameters.

In the equestrian world, two different types of management can be distinguished: traditional management and natural boarding. The aim of this research was to compare hormonal, hematological and immunological parameters of 47 horses kept in these two different managements. Blood and horsehair of the horses were sampled to determine DHEA (dehydroepiandrosterone) and cortisol concentration through RIA. Moreover, blood count was conducted, and flow cytometry was employed to phenotype lymphocyte subpopulations. Results showed that, in horsehair, DHEA concentration was significantly higher in natural horses, whereas cortisol concentration and cortisol/DEHA ratio significantly lower. These hormonal parameters are used to assess the stress condition and the welfare of animals. The most favorable endocrine framework found in horses kept in natural boarding suggests that this management conveys most with ethological and physiological needs of the species. The research underlines the need of a modification of horses' husbandry systems. For the first time, this study validates the assay of DHEA in horsehair.

Gum feeder as environmental enrichment for zoo marmosets and tamarins.

Tamarins and marmosets are small-bodied social callitrichines. Wild callitrichines feed on exudates, such as sap and gum; particularly, marmosets are mainly gummivores, while tamarins consume gums only occasionally and opportunistically. Zoo marmosets and tamarins are usually provided with gum arabic as an alternative to the exudates normally found in the wild. The aim of this study was to evaluate the effects of a gum feeder on the behavior and well-being of four zoo-managed callitrichines. We studied four cotton-top tamarins (Saguinus oedipus), four red-handed tamarins (S. midas), two pygmy marmosets (Cebuella pygmaea), and three Geoffroy's marmosets (Callithrix geoffroyi) housed at Parco Natura Viva (Italy). We conducted the study over two different periods, a baseline (control, without the gum feeder) and then a gum feeder (when the gum feeder was provided) period. We used continuous focal animal sampling to collect behavioral data, including durations of social and individual behaviors. We collected 240 min of observations per period per study subject, with a total of 3,120 min for all the subjects in the same period and of 6,240 min in both periods. We analyzed data by using nonparametric statistical tests. First, we found that the gum feeder promoted species-specific behaviors, such as exploration, and diminished self-directed behaviors, suggesting an enriching effect on tamarin and marmoset behavior. Moreover, in red-handed tamarins, the provision of the gum feeder reduced the performance of self-directed and abnormal behavior, specifically coprophagy. These results confirm that gum feeders are effective foraging enrichment tools for zoo marmosets and tamarins.

Role of gut microbiota in dog and cat's health and diseases.

Mammalian gastrointestinal tract is colonized by a large number of microorganisms, known as gut microbiota, that play a key role in the physiological and pathological states. In particular, the gastrointestinal tract of dogs and cats harbors a complex and highly biodiverse microbial ecosystem. Recent studies see it involved in a wide range of life processes, including energy needs, metabolism, immunological activity, and neuro-behavioral development. This review focuses on the role of the microbiota on the health of pets and will discuss changes that occur in the disease.

Fecal concentrations of cortisol, testosterone, and progesterone in cotton-top tamarins housed in different zoological parks: relationships among physiological data, environmental conditions, and behavioral patterns.

The aim of this investigation was to study the welfare of 3 captive groups of cotton-top tamarins housed in different zoological parks. Ethological observations were conducted during 1 year. In addition, fecal samples were collected and the concentrations of glucocorticoids, androgens, and progestogens were measured. Within each group, no significant differences in fecal cortisol concentrations were found between subjects. The fecal concentrations of testosterone and progesterone significantly differed depending on the sexes and ages of the tamarins. A significant association was found among hormone concentrations, exhibit dimensions, and group composition. A highly significant correlation was found between all hormones considered and the space available for each subject. Significant differences in behavioral patterns were observed among groups, including social-individual, affiliative-aggressive, and anogenital-suprapubic scent marking. Correlations between hormone measurements and behaviors were detected. In conclusion, this study confirmed the associations between some behaviors exhibited by these nonhuman primates and both cortisol and testosterone; these data also highlight the role played by progesterone in these behaviors.

Effect of ß-carotene Supplementation on Italian Trotter Mare Peripartum.

When the mare's estrous cycle resumes in winter, the ß-carotene content of hay is depleted. Sixty Italian trotter mares were randomly assigned to a Control or a Treated Group. Treated Group received 1g/d synthetic ß-carotene for 15 days from parturition. Blood samples collected at parturition and on days 5, 10 and 15 after partum were analysed for ß-carotene, vitamins A, progesterone, 17 ß-estradiol, the energy parameters (glucose, cholesterol, NEFA), the protein profile (total protein, albumin, urea) and LDH. Some changes in these measures were attributable to treatment, which significantly affected ß-carotene and 17 ß-estradiol concentrations. A significant effect was also found on the resumption of estrous activity (χ(2) test=P<0.052).

Does training make you smarter? The effects of training on dogs' performance (Canis familiaris) in a problem solving task.

This study investigates the influence of training experiences on dogs' performance in a problem solving task, namely opening a box to obtain food. One hundred and eighteen dogs allocated to two different groups according to their training experience (no/basic training vs high level training) were tested. In each group the dogs saw the researcher manipulating either the paw-pad or the lid, prior to being allowed free access to the apparatus. No effect of the locus of manipulation was observed. However, there was a strong effect of training on the dogs' performance regardless of manipulation condition. Compared to untrained dogs, highly trained dogs were more successful in opening the box and spent significantly more time interacting with the apparatus; whereas untrained dogs spent significantly more time looking back at their owners and the researcher. These results indicate that high levels of training improve dogs' problem solving ability, with dogs appearing to be more proactive in the their interaction with novel objects.