The present study assessed if salivary crystallization pattern (ferning pattern formed as a result of the higher levels of salt content in the dried sample) could be used for estrus detection and for diagnosis of pregnancy/non-pregnancy in dairy cows. Saliva and blood samples were collected from non-pregnant cycling cows (Sahiwal breed; n = 20) on alternate days from the day of estrus till next estrus. Then, all the cows were inseminated and saliva and blood sampling were continued further for a period of 22 d post-insemination. Pregnancy diagnosis was carried out on day 45 post-insemination and eight cows were found to be pregnant. The salivary crystallization pattern and estradiol:progesterone ratio during estrous cycle and during pregnancy were compared among these cows. Six types of salivary crystallization patterns were discerned; distinct patterns such as branch-like, fern-like, fir-like and combinations of these. Fern-like pattern was observed in all the cows on the day of estrus (first measurement day) and furthermore, all of the cows that subsequently became pregnant had fern-like salivary crystallization pattern at the time of insemination. Saliva of all the pregnant cows showed branch-fir type of crystallization pattern on day 16 post-breeding while only 50% of non-pregnant cows showed this pattern on day 16 of estrous cycle. The appearance of fern-like pattern was positively and significantly related to estradiol:progesterone ratio (r = 0.86; P < 0.001). The findings were validated on a separate group of cycling cows (n = 32). We can conclude that salivary crystallization pattern might serve as a non-invasive and cost effective and easy-to-use cow-side tool for estrus detection and early pregnancy/non-pregnancy diagnosis in cows upon validation on a larger sample size.

Refinement techniques are being increasingly employed in all fields of animal research to try to ensure that the highest standards of welfare are upheld. This review concerns one of the main emerging techniques for the assessment of welfare itself, namely the non-invasive measurement of glucocorticoids (GCs) as indicators of stress. The paper is divided into three sections. The first discusses the relationship between GCs and stress. The second section considers whether factors other than stress are linked to rises in GCs, eg exercise, oestrus cycle and diet. The final part examines the reliability of the non-invasive techniques that measure GCs from samples of saliva and faeces. Although it is important to take into account some caveats associated with the methodologies employed, it is concluded, nevertheless, that these techniques can give accurate and reliable information regarding the welfare status of an individual or group of animals without the procedures themselves causing any kind of distress to the subjects.

The ‘coping hypothesis’ of stereotypic behaviour — that stereotypies are performed as a means of helping the animal to cope with its environment by reducing stress — was tested using two adult female margays (Leopardus wiedii), an endangered neotropical small cat species. Within-individual and between-individual comparisons of the duration of stereotypic pacing and glucocorticoid concentration, measured non-invasively in faeces, indicated that stereotypic pacing did not help the two margays to cope with their captive environment by reducing their physiological stress level. However, hiding appeared to serve as a coping function in the two margays, not by immediately reducing the faecal glucocorticoid concentration, but rather as a long-term effect.

The effects of non-invasive, non-convulsive electrical neuromodulation (NINCEN) on depression, anxiety and sleep disturbance are inconsistent in different studies. Previous meta-analyses on transcranial direct current stimulation (tDCS) and cerebral electrotherapy stimulation (CES) suggested that these methods are effective on depression. However, not all types of NINECN were included; results on anxiety and sleep disturbance were lacking and the influence of different populations and treatment parameters was not completely analyzed. We searched PubMed, Embase, PsycInfo, PsycArticles and CINAHL before March 2021 and included published randomized clinical trials of all types of NINCEN for symptoms of depression, anxiety and sleep in clinical and non-clinical populations. Data were pooled using a random-effects model. The main outcome was change in the severity of depressive symptoms after NINCEN treatment. A total of 58 studies on NINCEN were included in the meta-analysis. Active tDCS showed a significant effect on depressive symptoms (Hedges' g = 0.544), anxiety (Hedges' g = 0.667) and response rate (odds ratio = 1.9594) compared to sham control. CES also had a significant effect on depression (Hedges' g = 0.654) and anxiety (Hedges' g = 0.711). For all types of NINCEN, active stimulation was significantly effective on depression, anxiety, sleep efficiency, sleep latency, total sleep time, etc. Our results showed that tDCS has significant effects on both depression and anxiety and that these effects are robust for different populations and treatment parameters. The rational expectation of the tDCS effect is ‘response’ rather than ‘remission’. CES also is effective for depression and anxiety, especially in patients with disorders of low severity.

Infertility is an important reproductive health problem, and male infertility is especially important in more than half of infertility cases. Due to the importance of genetic factors in this condition, analysis of semen alone is not enough to recognize men with idiopathic infertility. A molecular non-invasive investigation is necessary to gain valuable information. Currently, microRNAs (miRNAs) are being used as non-invasive diagnostic biomarkers. miRNAs, single-stranded non-coding RNA molecules, act as post-transcriptional gene silencing regulators either by inhibition or repression of translation. Changes in the regulation of miRNAs have been investigated in several different types of male infertility, therefore the biological role of miRNA and gene targets has been defined. The purpose of this study was to review recent research on the altered expression of miRNA in semen, sperm, and testicular biopsy samples in infertile males with different types of unexplained infertility. Changes in miRNA regulation were investigated using microarray and the miRNA levels were confirmed by real-time qRT-PCR. This review explains why creating a non-invasive diagnostic method for male infertility is necessary and how changes in miRNA expression can be used as new diagnostic biomarkers in patients with differing spermatogenic and histopathologic injury.

Current techniques for measuring the dry matter intake (DMI) of grazing lactating beef cows are invasive, time consuming and expensive making them impractical for use on commercial farms. This study was undertaken to explore the potential to develop and validate a model to predict DMI of grazing lactating beef cows, which could be applied in a commercial farm setting, using non-invasive animal measurements. The calibration dataset used to develop the model was comprised of 94 measurements recorded on 106 beef or beef–dairy crossbred cows (maternal origin). The potential of body measurements, linear type scoring, grazing behaviour and thermal imaging to predict DMI in combination with known biologically plausible adjustment variables and energy sinks was investigated. Multivariable regression models were constructed for each independent variable using SAS PROC REG and contained milk yield, BW, parity, calving day and maternal origin (dairy or beef). Of the 94 variables tested, 32 showed an association with DMI (P < 0.25) upon multivariable analysis. These variables were incorporated into a backwards linear regression model using SAS PROC REG. Variables were retained in this model if P < 0.05. Five variables; width at pins, full body depth, ruminating mastications, central ligament and rump width score, were retained in the model in addition to milk yield, BW, parity, calving day and maternal origin. The inclusion of these variables in the model increased the predictability of DMI by 0.23 (R2 = 0.68) when compared to a model containing milk yield, BW, parity, calving day and maternal origin only. This model was applied to data recorded on an independent dataset; a herd of 60 lactating beef cows two years after the calibration study. The R2 for the validation was 0.59. Estimates of DMI are required for measuring feed efficiency. While acknowledging challenges in applicability, the findings suggest a model such as that developed in this study may be used as a tool to more easily and less invasively estimate DMI on large populations of commercial beef cows, and therefore measure feed efficiency.

In this study, a video-based infrared camera (IRC) was investigated as a tool to monitor the body temperature of calves. Body surface temperatures were measured contactless using videos from an IRC fixed at a certain location in the calf feeder. The body surface temperatures were analysed retrospectively at three larger areas: the head area (in front of the forehead), the body area (behind forehead) and the area of the entire animal. The rectal temperature served as a reference temperature and was measured with a digital thermometer at the corresponding time point. A total of nine calves (Holstein-Friesians, 8 to 35 weeks old) were examined. The average maximum temperatures of the area of the entire animal (mean±SD: 37.66±0.90°C) and the head area (37.64±0.86°C) were always higher than that of the body area (36.75±1.06°C). The temperatures of the head area and of the entire animal were very similar. However, the maximum temperatures as measured using IRC increased with an increase in calf rectal temperature. The maximum temperatures of each video picture for the entire visible body area of the calves appeared to be sufficient to measure the superficial body temperature. The advantage of the video-based IRC over conventional IR single-picture cameras is that more than one picture per animal can be analysed in a short period of time. This technique provides more data for analysis. Thus, this system shows potential as an indicator for continuous temperature measurements in calves.

Monitoring rare and elusive species for effective management and conservation is particularly challenging and often demands the development of specialized techniques. Scat surveys have been applied to monitor a variety of rare species but relatively little attention has been given to the development of appropriate sampling designs. To determine if scat surveys could be applied to compare the distribution of species across three habitats of a fragmented region in the Brazilian Amazon, the removal of human (n = 27) and jaguar (n = 27) scat samples in forest, riparian corridor and pasture habitats was recorded for 24 hours. Dung beetles were responsible for removing the majority of samples (71%) and a generalized linear mixed effect model revealed significant influence of habitat and scat type on removal probability, with forest and riparian corridors having higher removal compared with samples in pasture habitats. Although non-invasive scat surveys can potentially address fundamental broad-scale conservation and management questions, our results demonstrate that scat surveys in the tropics must account for differences in scat removal rates between habitats and target species before conclusions can be drawn regarding patterns of habitat use.