Recognition of spatial parameters by echolocation in the vampire bat,Desmodus rotundus

Summary Three vampire bats (Desmodus rotundus) were trained in a two-choice apparatus (Fig. 1) to select the one of two passages that contained a vertical stick or narrow metal strip at a distance of 45 cm. To single sticks in isolation they responded at a width of 1.1 mm or more. Large surfaces in the vicinity of the target impaired the locating ability; the animals correctly choose a 20-mm-wide strip 2.5 cm in front of a strongly reflecting plate (Fig. 2) or 6 cm away on one side of it (Fig. 3).In a flight tunnel the response to single vertical obstacles of various size was studied. The sudden increase in echolocation-sound repetition rate as the obstacle was approached marked the response distance. This distance was 50–80 cm for thread obstacles 0.5 mm thick; the threshold diameter was 0.23 mm (Fig. 6). When the obstacles were mounted in a 30-cm-wide constriction of the tunnel the response occurred later; even a 10-mm-wide strip did not elicit a change in the sound pattern until the bats had approached to a distance of 60 cm (Fig. 6). The response distance increased significantly when the obstacles were moved in front of or beyond the constriction (Fig. 8).     

Poikilothermic traits in Mashona mole-rat (Fukomys darlingi). Reality or myth?

The African mole-rats (Bathyergidae, Rodentia) is a mammalian family well known for a variety of ecophysiological adaptations for strictly belowground life. The smallest bathyergid, the hairless naked mole-rat from arid areas in Eastern Africa, is even famous as the only truly poikilothermic mammal. Another bathyergid, the Mashona mole-rat (Fukomys darlingi) from Zimbabwe, is supposed to have strong poikilothermic traits, because it is not able to maintain a stable body temperature at ambient temperatures below 20 °C. This is surprising because, compared to the naked mole-rat, this species, together with all congenerics, is larger, haired, and living in more seasonal environment. In addition, other Fukomys mole-rats show typical mammalian pattern in resting metabolic rates. In our study, we measured resting metabolic rate and body temperature of Mashona mole-rats from Malawi across a gradient of ambient temperatures to test its poikilothermic traits. We found that the adult mass specific resting metabolic rate was 0.76±0.20 ml O₂ g⁻¹ h⁻¹ and body temperature 34.8±1.1 °C in the thermoneutral zone (27–34 °C). Body temperature was stable (33.0±0.5 °C) at ambient temperatures from 10 to 25 °C. We thus cannot confirm poikilothermic traits in this species, at least for its Malawian population. Factors potentially explaining the observed discrepancy in Mashona mole-rat energetics are discussed.     

Electric field perturbations of spiral waves attached to millimeter-size obstacles

Reentrant spiral waves can become pinned to small anatomical obstacles in the heart and lead to monomorphic ventricular tachycardia that can degenerate into polymorphic tachycardia and ventricular fibrillation. Electric field-induced secondary source stimulation can excite directly at the obstacle, and may provide a means to terminate the pinned wave or inhibit the transition to more complex arrhythmia. We used confluent monolayers of neonatal rat ventricular myocytes to investigate the use of low intensity electric field stimulation to perturb the spiral wave. A hole 2-4 mm in diameter was created in the center to pin the spiral wave. Monolayers were stained with voltage-sensitive dye di-4-ANEPPS and mapped at 253 sites. Spiral waves were initiated that attached to the hole (n = 10 monolayers). Electric field pulses 1-s in duration were delivered with increasing strength (0.5-5 V/cm) until the wave terminated after detaching from the hole. At subdetachment intensities, cycle length increased with field strength, was sustained for the duration of the pulse, and returned to its original value after termination of the pulse. Mechanistically, conduction velocity near the wave tip decreased with field strength in the region of depolarization at the obstacle. In summary, electric fields cause strength-dependent slowing or detachment of pinned spiral waves. Our results suggest a means to decelerate tachycardia that may help to prevent wave degeneration.     

Locomotor Adaptation versus Perceptual Adaptation when Stepping Over an Obstacle with a Height Illusion

Background

During locomotion, vision is used to perceive environmental obstacles that could potentially threaten stability; locomotor action is then modified to avoid these obstacles. Various factors such as lighting and texture can make these environmental obstacles appear larger or smaller than their actual size. It is unclear if gait is adapted based on the actual or perceived height of these environmental obstacles. The purposes of this study were to determine if visually guided action is scaled to visual perception, and to determine if task experience influenced how action is scaled to perception.

Methodology/Principal Findings

Participants judged the height of two obstacles before and after stepping over each of them 50 times. An illusion made obstacle one appear larger than obstacle two, even though they were identical in size. The influence of task experience was examined by comparing the perception-action relationship during the first five obstacle crossings (1–5) with the last five obstacle crossings (46–50). In the first set of trials, obstacle one was perceived to be 2.0 cm larger than obstacle two and subjects stepped 2.7 cm higher over obstacle one. After walking over the obstacle 50 times, the toe elevation was not different between obstacles, but obstacle one was still perceived as 2.4 cm larger.

Conclusions/Significance

There was evidence of locomotor adaptation, but no evidence of perceptual adaptation with experience. These findings add to research that demonstrates that while the motor system can be influenced by perception, it can also operate independent of perception.     

Differential responses of the floating-leaved aquatic plant Nymphoides peltata to gradual versus rapid increases in water levels

We compared the growth responses of the floating-leaved species Nymphoides peltata to gradual and rapid rising water levels under two nutrient concentrations (1 g and 12 g of slow released fertilizer (N-P-K: 16-8-12) per container filled with 8 kg washed sand), and predicted the population expansion after these floods. The results showed that the capacity for petiole elongation was dependent on leaf age, and only leaves that were no more than five days old had the capability to reach the water surface when the water level increased rapidly from 50 cm to 300 cm. Plants subjected to a gradual rising water level tracked the increase in water depth whose petioles elongated at 3.96 ± 1.70 cm per day and 4.80 ± 0.16 cm per day under low and high nutrient concentrations respectively throughout the experiment period. When water levels were rapidly raised, leaf petioles elongated rapidly at 25.48 ± 1.51 cm per day and 26.64 ± 2.24 cm per day under low and high nutrient concentrations respectively during the first ten days. Plants under a constant water level maintained highest mean leaf recruitment (mean 3.0 ± 0.33 leaves and 24.4 ± 5.87 leaves every ten days under low and high nutrient concentrations, respectively). Therefore, more young leaves existed in the canopy ensuring that when the water level increases, young leaves can rapidly emerge after submergence. Gradual water level rise did not significantly affect biomass and ramet production (4.75 ± 1.41 g and 5.50 ± 1.22 ramets in low nutrient; 48.49 ± 21.45 g and 35.67 ± 11.78 ramets in high nutrient), but rapid water level rise negatively affected ramet production in both nutrient concentrations (3.00 ± 1.26 ramets and 11.25 ± 4.19 ramets in low and high nutrients, respectively). The results indicated that continual leaf recruitment and rapid petiole elongation were both important ways in which N. peltata adapted to increasing water levels. Extreme flooding may be a disturbance factor that affects plant growth and the population expansion of N. peltata, while small gradual water level rise should not harm this species.     

Germination and early growth of Nymphaea odorata at different water depths

We experimentally determined the effects of water depth on seed germination and seedling growth and morphology, and we documented the transition from submerged to emergent plants in the white water lily, Nymphaea odorata. Seeds of N. odorata were germinated at 30, 60, and 90 cm water depth in outdoor mesocosms and percent germination and morphology measured after a month. The presence of self-seeded seedlings in pots at the same 3 water levels was also recorded over two years. To examine juvenile growth, seeds planted in soil were placed at the same mesocosm depths; germination and growth were monitored for three months, when the plants were harvested for morphological and biomass measurements. N. odorata germinated equally well in 30, 60 and 90 cm water; seedlings grew as submerged aquatics. After one month, seedlings in 90 cm water had less biomass than those in 30 cm (1.1 vs. 3.3 mg and 1.0 vs. 1.8 mg for different seed sources, respectively) and allocated relatively more biomass to shoots (97.5 vs. 67.8% and 73.1 vs. 58.0%, respectively). Seedlings in 60 cm water were intermediate. After 3 months of submerged growth, plant biomass remained less in 90 vs. 60 and 30 cm water (22.5 vs. 36.4 and 33.3 mg, respectively). Plants in 90 and 60 cm water had greater biomass allocation to shoots than plants in 30 cm water (85.7 and 72.6% vs. 64.4%, respectively) and produced larger laminae on longer petioles (lamina length = 33.3 vs. 25.2 mm in 90 vs. 30 cm; petiole length = 99.0 vs. 36.0 mm, respectively). After about 3 months, submerged plants produced floating leaves that had 39% shorter laminae but 267% to 1988% longer petioles than submerged leaves on the same plant. Lamina length to width allometric relations of submerged leaves were >1 at all water levels, distinguishing them from the equal allometry of adult floating leaves. The switch from production of submerged to emergent leaves resembles submergence-escape growth in other aquatics, but because the seedlings have been submerged throughout their life, submergence itself cannot be the stimulus to produce emergent leaves in these totally immersed plants. Our data show that N. odorata plants can establish from seeds in up to 90 cm water and that seedlings grow as submerged aquatics until they switch abruptly to production of floating leaves.     

Inconsistent effects of reefs on different size classes of macrofauna in adjacent sand habitats

In soft-sediment communities near reefs, a variety of patterns have been described with distance from the reef edge. Various studies have observed contrasting patterns and one study has reported different patterns for different size classes of macrofauna. This study in northeastern New Zealand obtained samples from 24 randomly allocated sites across three locations in a large-scale mensurative sampling design. At each location, there was a manipulation of reef-associated predator populations provided by established marine reserves. A concurrent study using the same sites found large macrofauna (> 4 mm) to vary with distance from the reef and relative to predator density. The present study sampled small-bodied infauna (< 4 mm and > 0.5 mm), which was also predicted to change with distance from the reef and predator density. In contrast to patterns found for larger fauna and to previous studies of small macrofauna, no consistent patterns were found for small-bodied infauna. These results suggest that models of community structure need to consider different size classes of macrofauna separately and that multiple sampling methods will assist their investigation. The ‘haloes’ in small-bodied macrofauna around reefs may not be as widespread or consistent as previous studies have suggested, particularly in mobile sediments on open coasts.     

The effect of temperature, water level and burial depth on seed germination of Myriophyllum spicatum and Potamogeton malaianus

The effects of temperature, water level and burial depth on seed germination of two submerged species, Myriophyllum spicatum and Potamogeton malaianus, were investigated under controlled laboratory conditions. There was no significant difference in final germination of M. spicatum among water level treatments, but P. malaianus germinations at 1 cm and 12 cm water levels were better than at 0 cm water level at temperatures of 20 °C and 30 °C. Little to no germination was observed for either species at the temperature of 10 °C. At 15 °C, however, germination increased significantly to 66.3-70.6% for M. spicatum and to 29.4-48.1% for P. malaianus under all three water level treatments. Increased temperature from 15 °C to 30 °C had no significant effect on the final germination of M. spicatum except at the 1 cm water level, but enhanced significantly the germination of P. malaianus. Analysis of the mean time to germination revealed that M. spicatum was a faster germinator relative to P. malaianus. The two species’ germination differed markedly in response to burial depth. Germination percentage of M. spicatum was 71.3% at 0 cm burial depth, but decreased to 5.0% and to 2.5% at depths of 1 cm and 2 cm, respectively; whereas germination percentages of P. malaianus were 40.0%, 23.8%, 12.5%, 7.5% and 1.3% at depths of 0 cm, 1 cm, 2 cm, 3 cm and 5 cm, respectively. We concluded that the two species respond differently to germination strategies. The findings provided further insight into how germination strategy contributes to the seed bank formation and species invasion.     

Variation in Arundo donax stem and leaf strength: Implications for herbivory

We determined leaf and stem strength for Arundo donax from plants grown in different geographic areas and at different times within their growing cycle. Mean leaf strength for plants collected within California was 1.72 Newtons (N) and ranged from 0.36 to 6.32 N, based on 1170 individual determinations. For leaves collected from 30 plants within four states between July 11 and 20, 2007, mean leaf strength was 1.58 N based on 936 determinations. Values ranged from 0.24 to 4.90 N. Overall, leaf strength showed statistically significant changes depending on the sampling date, sampling location, type of leaf sampled, and position within the leaf where the measurement was taken. In general leaf strength was greater near the base of the leaf and decreased with distance away from the base; leaf strength changed as the growing season progressed; and first year leaves had leaf strength values about 25% greater than leaves produced on stems >1-year old. This represents two of the three age categories of leaves which may be present on giant reed at any one time. Stem strength and stem wall thickness were strongly correlated (Kendall's Tau b = 0.92, P < 0.0001, N = 26). Linear regression indicated that mean stem strength decreased by approximately 6.8% (95% confidence limits 5.8-7.7%) from one node to the successive node progressing from the stem base to the shoot tip. These results imply that the ability of biological control agents to damage A. donax leaves may not be the same across the locations this plant occurs or at all times during the growing season within a given location.     

Successful homing of magnet-carrying white-chinned petrels released in the open sea

During the incubation period, white-chinned petrels, Procellaria aequinoctialis, repeatedly make foraging trips of several thousand kilometres in the open ocean from their small isolated breeding island. To test whether these birds rely on geomagnetic information to home, we displaced five individuals and released them in the open sea 300-360 km from home. They were prevented both from using route-based information during the passive displacement from the breeding island to the release site, and from using the geomagnetic field at the release site and on their way home (by carrying a mobile magnet on the head). They were equipped with satellite transmitters to record their homing movements. The five petrels homed efficiently (in 17-32 h) along paths that were on average slightly less than twice the homing distance. This showed that, at least when within a few hundred kilometres of home, white-chinned petrels are able to home successfully by relying only on nongeomagnetic site-dependent information. Copyright 2003 Published by Elsevier Science Ltd on behalf of The Association for the Study of Animal Behaviour.      

Prevention of floret abscission for Agapanthus praecox requires an adequate supply of carbohydrate to the developing florets

Vase solutions of 2.5 or 5% (w/v) sucrose markedly reduced the abscission of all open florets and buds on cut Agapanthus praecox stems. Pulsing cut stems with these solutions was not as effective as continuous treatments. Pulse treatments with 10% sucrose for 4–24 h were relatively ineffective at reducing floret and bud abscission but longer pulses of 48 h reduced abscission. The relative ineffectiveness of sucrose pulse treatments appeared to be due to the low water uptake of the stems (1–3 ml/day). Reducing the number of florets and buds on an inflorescence reduced abscission of florets and buds, and increasing stem length from 25 cm to 50 cm decreased bud abscission. This reduction in abscission is possibly due to the increased availability of assimilates for the remaining buds and florets or reduced competition for assimilates. Exogenous ethylene treatments (9 µl/L for up to 24 h) had no effect on abscission, although STS treatment (4 mM, 4 h) significantly reduced floret abscission when stems were held in vase solutions of sucrose. We conclude that postharvest floret abscission in A. praecox is influenced primarily by the availability of assimilates to the developing florets.     

Variation in the ability to communicate three-dimensional resource location by stingless bees from different habitats

We evaluated the ability of two Brazilian stingless bee species, Melipona mandacaia andM. bicolor , to recruit nestmates to a specific three-dimensional location. We used experimental feeder arrays and provide the first detailed evidence demonstrating that recruitment communication in Melipona can lead to large, rapid and highly significant increases in the number of nestmates visiting a specific location. Melipona bicolor andM. mandacaia foragers both recruited nestmates to the correct distance and direction, but differed in their ability to recruit nestmates to the correct height. These differences may relate to their respective habitats. Melipona mandacaia inhabits semi-arid areas of Caatinga where most food sources occur close to the ground, and its foragers evidently cannot recruit nestmates to the correct height. Melipona bicolor, an Atlantic rainforest species, evidently does not communicate height when the food source is at ground level, but can communicate height when the food source is at the forest canopy level (12 m high), where major food sources occur. Species-specific variation in three-dimensional location communication is intriguing because it suggests that Melipona may be a good model for studying the evolution of recruitment communication systems in highly social bees.     

Vertical fatty acid profiles in blubber of a freshwater ringed seal — Comparison to a marine relative

We investigated the vertical stratification pattern of fatty acids in the blubber of the freshwater Saimaa ringed seal (Phoca hispida saimensis; n = 35). Blubber was dissected vertically into 3 mm thick sequential subsamples, and the fatty acid composition was analyzed separately for each subsample. The combined vertical fatty acid profiles (expressed as numerical gradients) showed that the blubber of > 1 year old individuals is stratified into three distinguishable layers: superficial (~ 1.5 cm), middle and deep (~ 1 cm). Thickness of the middle layer varied according to the total blubber thickness. We suggest that the observed layering is related to the differences in the tissue temperatures and metabolic activity in the blubber column. The dissimilarity in the fatty acid composition (measured as Euclidean distance) between the different blubber layers and the 5 most important fish prey species available in Lake Saimaa was largest in the superficial blubber. In fact, the fatty acid composition of superficial blubber resembled more that of marine ringed seal (Phoca hispida hispida) blubber than any of the analyzed fish species. The composition closest to that of the prey was found in the deep blubber of the Saimaa ringed seal. The large vertical differences in the fatty acid composition of blubber lipids, which likely affect the vertical distribution of other endogenous or exogenous lipophilic substances as well, will set conditions for the sampling of blubber for biomonitoring and dietary studies. Thus the knowledge on the potential layers with different composition and the depths they span in different individuals (e.g. young versus old; with thin versus thick blubber) is crucial for improving the validity and reliability of monitoring methods utilizing the blubber tissue.     

Intracellular ciliated protozoal infection in silverlip pearl oysters, Pinctada maxima (Jameson, 1901)

The pathology associated with an intracellular ciliate infection in the digestive gland of pearl oysters Pinctada maxima (Jameson, 1901) is described. Histopathological and transmission electron microscopic examination were used to characterise the organism and its location within host cells. The parasite is tear-drop shaped measuring 5.53 μm (range of 2.73-7.47 μm, n = 9) in width and 11.15 μm (range of 9.02-16.2 μm) in length with a centrally located lobulated nucleus and a large nucleus:cytoplasmic ratio. The ciliate has nine evenly spaced rows of cilia running obliquely along the length of cell, converging on the pointed end. Infected digestive glands typically had a moderate to severe infiltration with mononuclear hemocyte. A strong correlation existed between the burden of ciliates and the host response; (p < 0.001, C = 0.315 Pearson Correlation). The use of a single tissue section upon microscopic examination was found to detect only 38-50% of the infections. However, examination of serial haematoxylin and eosin stained sections improved the reliability of detecting infection.     

Peripheral administration of oxytocin increases social affiliation in the naked mole-rat (Heterocephalus glaber)

The neuropeptide oxytocin regulates a wide variety of social behaviors across diverse species. However, the types of behaviors that are influenced by this hormone are constrained by the species in question and the social organization that a particular species exhibits. Therefore, the present experiments investigated behaviors regulated by oxytocin in a eusocial mammalian species by using the naked mole-rat (Heterocephalus glaber). In Experiment 1, adult non-breeding mole-rats were given intraperitoneal injections of either oxytocin (1 mg/kg or 10 mg/kg) or saline on alternate days. Animals were then returned to their colony and behavior was recorded for minutes 15–30 post-injection. Both doses of oxytocin increased huddling behavior during this time period. In Experiment 2, animals received intraperitoneal injections of either oxytocin (1 mg/kg), an oxytocin-receptor antagonist (0.1 mg/kg), a cocktail of oxytocin and the antagonist, or saline across 4 testing days in a counterbalanced design. Animals were placed in either a 2-chamber arena with a familiar conspecific or in a small chamber with 1 week old pups from their home colony and behaviors were recorded for minutes 15–30 post-injection. Oxytocin increased investigation of, and time spent in close proximity to, a familiar conspecific; these effects were blocked by the oxytocin antagonist. No effects were seen on pup-directed behavior. These data suggest that oxytocin is capable of modulating affiliative-like behavior in this eusocial species.     

Stimulus discrimination by horses under scotopic conditions

Scotopic vision in horses (Equus caballus) was investigated using behavioral measurements for the first time. Four horses were tested for the ability to make simple visual discriminations of geometric figures (circles and triangles) under various brightness levels within an enclosed building. Measurements of brightness ranging from 10.37 to 24.12 magnitudes per square arcsecond (mag/arcsec²; in candelas per square meter—7.70 to 2.43E-05 cd/m²) were taken using a Sky Quality Meter. These values approximated outdoor conditions ranging from twilight in open country to a dark moonless night in dense forest. The horses were able to solve the discrimination problems in all brightness settings up to 23.77 mag/arcsec² (3.35E-05 cd/m²). Moreover, they easily navigated their way around obstacles located within the testing area in extremely dim light (>23.50 mag/arcsec²; 4.30E-05 cd/m²), which were in conditions too dark for the human experimenters to see. These findings support physiological data that reveal a rod-dominated visual system as well as observations of equine activity at night.     

Water quality effectiveness of ditch fencing and culvert crossing in the Lake Okeechobee basin, southern Florida, USA

Ditch fencing and culvert cattle crossing Best Management Practice (BMP) was evaluated in this study with regard to phosphorus (P) and nitrogen (N) load reductions and economic feasibility in the Lake Okeechobee (LO) basin. The BMP was implemented at a 170 m section of a drainage ditch within a ranch in the LO basin and flow and concentration (N and P) data at the upstream and downstream of the ditch were collected for one pre-BMP (June-October, 2005) and three post-BMP (June-October, 2006-2008) periods. During the pre-BMP period, downstream total P (TP) load was 20% (67.0 kg) higher than the upstream, indicating the cattle crossing to be a source of P. Downstream loads of TP in 2006 and 2008 (post-BMP periods) became 26% (14.7 kg) and 11% (85.9 kg) lower than the upstream loads, respectively indicating that the BMP reduced the P loads. The site was a sink for N for all periods except the 2007. Unusual dry conditions during 2007 resulted in the addition of P and N at the BMP site, probably due to the release of P and N from soil and plants. Average of three post-BMP period load showed a 10% reduction of TP loads at the downstream (251.8 kg) compared to the upstream (281.0 kg) location. To consider potential P contributions from the soil and plant, two scenarios, conservative and liberal, were considered to estimate P load reductions due to the BMP. For the conservative scenario, P contribution from soil and plant was considered, while for liberal it was not. Reductions in P loads for conservative and liberal scenarios were 0.35 and 0.44 kg/day, respectively. Phosphorus removal cost for the conservative scenario was $12.61/kg of P, which is considerably less than the cost of other P reduction strategies in the basin. Overall, results show that the BMP can reduce P concentration and loads from ranches without causing adverse impact on cattle production.     

A bis(chlorido)-bridged linear-chain Cu(II) compound with 7-azaindole as an axial ligand; synthesis, structure, hydrogen bonding and magnetism

A novel linear bis(chlorido)-bridged Cu(II) compound with the ligand 7-azaindole (abbreviated as Haza) have been synthesized and fully characterized by spectroscopy, EPR and X-ray structure analysis. The geometry around the Cu(II) ion in the compound [Cu-μ-Cl₂(Haza)₂]_n, is distorted octahedral with the basal plane formed by two chloride anions with a Cu-Cl distance of 2.286(1) Å and two nitrogen atoms of two trans-chelating Haza ligands with a Cu-N distance of 2.042(4) Å. The apical positions is formed by two chloride anions of a neighbouring unit at a distance of 3.143(2) Å. The Cu-Cl-Cu angle is 90.09(5)°, while the Cu-Cu distance is 3.8890(8) Å.The EPR powder spectrum appears as rhombic, displaying some extra features which were assigned to the differences in orientation of chains in the lattice, as their intensity appears to decrease on powdering. The magnetic susceptibility measurements, recorded from 5 to 300 K, agree with a very weak antiferromagnetic interaction in a chain with J = −2.6 cm⁻¹.     

Long-term stability of Avoidance Distance tests for on-farm assessment of dairy cow relationship to humans in alpine traditional husbandry systems

The present study investigates the long-term stability of two behaviour tests used to evaluate the human-animal relationship as well as gather information to refine on-farm welfare assessment protocols in alpine husbandry systems. Two tests, Avoidance Distance (AD) and Avoidance Distance at the Feeding rack (ADF) were performed on 50 lactating cows, bred in a loose housing system, following a protocol validated for dairy cows in the Welfare Quality^® project. AD was performed on each animal at three different times: at the end of the indoor housing period just before the grazing period (AD_bg); during the outdoor summer grazing period in mountain ranges (AD_dg); and at the beginning of the indoor housing period, just after the grazing period (AD_ag). ADF was performed on the same cows only during the housing period, at the bottom of the valley, before (ADF_bg) and after (ADF_ag) the grazing period. Values of AD_ag (113.93 ± 21.21 cm) were significantly higher (P < 0.05) than those observed in AD_bg (71.07 ± 14.63 cm) and in AD_dg (77.68 ± 18.11 cm). No statistical significance was pointed out between AD_bg and AD_dg. A similar trend was recorded also for ADF, which was significantly higher after the grazing period (ADF_bg vs ADF_ag: 25.00 ± 6.80 vs 47.50 ± 8.72 cm; P < 0.05). The summer grazing period seems to affect the reaction of cows towards humans, and AD and ADF are not consistent throughout the year. Certification of on-farm welfare in alpine husbandry systems, therefore requires performing these tests at the end of the indoor period, to gain better insight of the human-animal relationship for the environment in which cows live most of the year.