3D pose estimation to detect posture transition in free-stall housed dairy cows.

Free stall comfort is reflected in various indicators, including the ability for dairy cattle to display unhindered posture transition movements in the cubicles. To ensure farm animal welfare, it is instrumental for the farm management to be able to continuously monitor occurrences of abnormal motions. Advances in computer vision have enabled accurate kinematic measurements in several fields such as human, equine and bovine biomechanics. An important step upstream to measuring displacement during posture transitions is to determine that the behavior is accurately detected. In this study, we propose a framework for detecting lying to standing posture transitions from 3D pose estimation data. A multi-view computer vision system recorded posture transitions between Dec. 2021 and Apr. 2022 in a Swedish stall housing 183 individual cows. The output data consisted of the 3D coordinates of specific anatomical landmarks. Sensitivity of posture transition detection was 88.2% while precision reached 99.5%. Analyzing those transition movements, breakpoints detected the timestamp of onset of the rising motion, which was compared with that annotated by observers. Agreement between observers, measured by intra-class correlation, was 0.85 between 3 human observers and 0.81 when adding the automated detection. The intra-observer mean absolute difference in annotated timestamps ranged from 0.4s to 0.7s. The mean absolute difference between each observer and the automated detection ranged from 1.0s to 1.3s. There was a significant difference in annotated timestamp between all observer pairs but not between the observers and the automated detection, leading to the conclusion that the automated detection does not introduce a distinct bias. We conclude that the model is able to accurately detect the phenomenon of interest and that it is equatable to an observer.

The effect of weaning age on animal performance in lambs exposed to naturally acquired nematode infections.

The effects of mixed gastrointestinal nematode (GIN) infections on animal growth and post-weaning activity patterns were investigated in grazing intact ram lambs when naturally exposed to two different infection levels and weaned at different ages. Ewes and their twin-born lambs were turned-out to graze in two permanent pasture enclosures naturally contaminated with GIN the previous year. Ewes and lambs in the low parasite exposure group (LP) were drenched before turn-out and at weaning, respectively, with 0.2 mg ivermectin per kg body weight, whereas those in the high parasite exposure group (HP) were left untreated. Two weaning ages were applied, early weaning (EW) (10 weeks) and late weaning (LW) (14 weeks), respectively. The lambs were then allocated to one out of four groups based on parasite exposure level and weaning age (EW-HP, n = 12; LW-HP, n = 11; EW-LP, n = 13; LW-LP, n = 13). Body weight gain (BWG) and faecal egg counts (FEC) were monitored, in all groups, from the day of early weaning and every four weeks, for 10 weeks. In addition, nematode composition was determined using droplet digital PCR. Activity patterns measured as Motion Index (MI; the absolute value of the 3D acceleration) and lying time were monitored continuously from the day of weaning until four weeks post-weaning using IceQube® sensors. Statistical analyses were performed in RStudio, using mixed models with repeated measures. BWG was 11% lower in EW-HP compared with EW-LP (P = 0.0079) and 12% lower compared with LW-HP (P = 0.018), respectively. In contrast, no difference in BWG was observed between LW-HP and LW-LP (P = 0.97). The average EPG was higher in EW-HP compared with EW-LP (P < 0.001), as well as in EW-HP compared with LW-HP (P = 0.021), and LW-HP compared with LW-LP (P = 0.0022). The molecular investigation showed that animals in LW-HP had a higher proportion of Haemonchus contortus compared with EW-HP. MI was 19% lower in EW-HP compared with EW-LP (P = 0.0004). Daily lying time was 15% shorter in EW-HP compared with EW-LP (P = 0.0070). In contrast, no difference in MI (P = 0.13) and lying time (P = 0.99) between LW-HP and LW-LP was observed. The results suggest that a delayed weaning age may reduce the adverse effects of GIN infection on BWG. Contrarily, an earlier weaning age may reduce the risk of H. contortus infection in lambs. Moreover, the results demonstrates a potential use of automated behaviour recordings as a diagnostic tool for the detection of nematode infections in sheep.

Intestinal ischemia measured by intraluminal microdialysis.

OBJECTIVE: To evaluate the possibility of detecting intestinal ischemia by intraluminal microdialysis and comparing the ileum and colon. METHODS: The studies were performed on male Sprague-Dawley rats. In the first part of the study, microdialysis catheters were placed in the sigmoid part of the colon and in the subcutaneous adipose tissue. In the second part of the study, microdialysis catheters were placed in the lumen of the ileum and the colon. The infrarenal aorta was clamped proximal to the cranial mesenteric artery. Microdialysate levels of glucose, lactate, pyruvate and glycerol were measured. Intestinal specimens were removed at the end of the ischemic period for microscopic evaluation. RESULTS: Intraluminal microdialysis could detect early signs of ischemic injury in the ileum, as well as in the colon, with a marked increase of lactate, lactate/pyruvate ratio and glycerol. The increased levels of intraluminal glycerol showed a positive correlation to prolonged ischemia and to higher degrees of intestinal damage. CONCLUSION: Intraluminal measurement of glycerol is a good marker for intestinal ischemia. Intraluminal microdialysis in the colon is easily accessible through the rectum, and may prove to be a valuable clinical tool for diagnosing intestinal ischemia.

Assessment of three DNA extraction kits for the absolute quantification of strongyle nematode eggs in faecal samples.

BACKGROUND: Haemonchus contortus is one of the most pathogenic gastrointestinal nematodes of small ruminants. The current diagnostic approach for the detection of this species relies on coproscopic methods, which both have low sensitivity and are time consuming. Methods employing detection through DNA amplification, such as droplet digital polymerase chain reaction (ddPCR), offer an advantageous approach to the diagnosis of H. contortus. However, DNA extraction protocols need to be constantly updated for the optimal retrieval of diagnostically usable template. Here, we describe the evaluation of three genomic DNA extraction kits for the detection and quantification of H. contortus ITS2 amplicon DNA from faecal samples, using droplet digital PCR. RESULTS: DNA samples, extracted from faecal material with the Nucleospin DNA Stool kit, produced the highest amounts of ITS2 amplicon copies and had the lowest coefficient of variation across different dilutions and sample types (fresh or frozen) out of the tested kits (Nucleospin DNA Stool, E.Z.N.A.® Stool DNA Kit and QIAamp Fast DNA Stool Mini Kit). Furthermore, the protocol of this kit has the fewest number of steps and the price of DNA extraction per sample is reasonable (2.77 €). CONCLUSIONS: The Nucleospin DNA Stool kit is an attractive option for the detection and quantification of H. contortus DNA in faecal samples of small ruminants in a diagnostic setting.

Percutaneous hernia repair in children: Safe to introduce.

BACKGROUND AND OBJECTIVE: In 2014 we introduced percutaneous internal ring suture as an alternative to open surgery for the treatment of inguinal hernia. This study aims to evaluate the introduction of the procedure at our institution. METHODS: In total, 100 consecutive patients operated with percutaneous internal ring suture were compared with 100 consecutive patients operated with open surgery. The patients were operated from August 2014 until November 2017. Patient demographics, clinical history, operative time, time in theater, and postoperative complications were extracted from charts. RESULTS: The mean operative time for percutaneous internal ring suture was 26.54 min and for open surgery 39.94 min, P < 0.0001. The total mean operative theater time for percutaneous internal ring suture was 108.95 min and for open surgery 118.4 min, P = 0.0343. During follow-up, two percutaneous internal ring sutures were operated for recurrent hernia. In the open surgery-group, three patients were operated for recurrent hernia, three for secondary testicular retention, and three for metachronous contralateral hernia. CONCLUSIONS: Even when established as a new technique, the percutaneous internal ring suture procedure is safe and results in shorter operative time and shorter theater time compared to open surgery.

Nematode parasitism affects lying time and overall activity patterns in lambs following pasture exposure around weaning.

We investigated the effects of gastrointestinal nematode (GIN) challenge on activity in first season grazing lambs naturally exposed to two different levels of multispecies GIN infections. Ewes and their twin-born lambs were turned-out to graze in two permanent pasture enclosures naturally contaminated with GIN the previous year, thereby exposing them to overwintering strongyle larvae. Animals in the low parasite exposure group (LP) were dewormed monthly with 0.2 mg ivermectin (Ivomec® vet, oral suspension) per kg body weight, whereas those in high parasite exposure group (HP) were left untreated. At weaning, lambs were allocated to one out of four groups based on weight and sex (HPE, n = 15; HPR, n = 15; LPE, n = 14; LPR, n = 14), in four nearby non-contaminated ley enclosures of similar size. Activity patterns were monitored from day -7, i.e. 7 days pre-weaning, until day 49, i.e. 49 days post-weaning, by fitting all lambs with IceQube sensors (IceRobotics). Body weight was monitored weekly from day -21, whereas faecal samples were investigated at days -21, 7, 35 and 49 for nematode faecal egg counts (EPG) using McMaster-technology and a validated Droplet Digital PCR protocol to determine nematode composition. All statistical analyses were performed in R studio, using mixed models with repeated measures. In the data analyses, weekly recordings was treated as a period, generating a total of eight periods. Average daily lying time had a significant interaction between parasite exposure and period (P = 0.0013), with animals in HP having a 101 ± 31 min shorter daily lying time compared to LP. Motion Index (MI; absolute value of the 3-D acceleration) had a significant interaction between parasite exposure and period (P = 0.0001) with lambs in group HP having a lower average daily MI compared with LP. Both body weight gain and EPG levels were significantly different (P<0.0001) between HP and LP groups during the course of the study. The molecular investigation showed that animals were predominantly infected with Teladorsagia spp., combined with low proportions of Haemonchus spp. In conclusion, this study shows that lying time and Motion Index of lambs around weaning was affected by moderate nematode infections. This indicates that there is a potential use of automated behaviour recordings as a diagnostic tool for detection of nematode parasites in lambs even at moderate infection levels.

Animal Welfare Implications of Digital Tools for Monitoring and Management of Cattle and Sheep on Pasture.

The opportunities for natural animal behaviours in pastures imply animal welfare benefits. Nevertheless, monitoring the animals can be challenging. The use of sensors, cameras, positioning equipment and unmanned aerial vehicles in large pastures has the potential to improve animal welfare surveillance. Directly or indirectly, sensors measure environmental factors together with the behaviour and physiological state of the animal, and deviations can trigger alarms for, e.g., disease, heat stress and imminent calving. Electronic positioning includes Radio Frequency Identification (RFID) for the recording of animals at fixed points. Positioning units (GPS) mounted on collars can determine animal movements over large areas, determine their habitat and, somewhat, health and welfare. In combination with other sensors, such units can give information that helps to evaluate the welfare of free-ranging animals. Drones equipped with cameras can also locate and count the animals, as well as herd them. Digitally defined virtual fences can keep animals within a predefined area without the use of physical barriers, relying on acoustic signals and weak electric shocks. Due to individual variations in learning ability, some individuals may be exposed to numerous electric shocks, which might compromise their welfare. More research and development are required, especially regarding the use of drones and virtual fences.

Subclinical nematode parasitism affects activity and rumination patterns in first-season grazing cattle.

Sickness behaviour has been suggested as an applicable indicator for monitoring disease. Deviating feeding behaviour and activity can provide information about animals' health and welfare status. Recent advances in sensor technology enable monitoring of such behaviours and could potentially be utilized as an indicator of gastrointestinal nematode (GIN) infections. This study investigated activity and rumination patterns in first-season grazing steers exposed to subclinical infection levels of the GIN Ostertagia ostertagi and Cooperia oncophora. At turnout, animals were allocated to one of four experimental groups and were faced with "high" (H1, n = 15; H2, n = 17) or "low" (L1, n = 17; L2, n = 11) levels of parasite exposure by grazing in similar enclosures contaminated with overwintering third stage (L3) GIN larvae. Animals in H1 and H2 (HP) received a 1:1 mix of approximately 10,000 O. ostertagi and C. oncophora L3 at turnout; whereas the animals in L1 and L2 (LP) were treated monthly with ivermectin. Activity and rumination patterns were monitored by fitting animals with leg- (IceQube) and neck-mounted (Heatime) sensors. BW was recorded every fortnight, whereas faecal and blood samples were examined every four weeks for nematode faecal egg count and serum pepsinogen concentrations (SPCs). There was an interaction effect of exposure level and period (P < 0.0001) on average lying daily time across the entire grazing time. A higher mean daily lying time (P = 0.0037) was found in HP compared with LP during the first 40 days on pasture. There was also interaction effects of treatment and day since turnout on rumination time (P < 0.0001) and rumination change (P = 0.0008). Also mean daily steps (P < 0.0001) and mean daily motion index (P < 0.0001) were markedly higher in HP during days 62-69, coinciding with peaking SPC in HP. Strongyle eggs were observed both in HP and LP from 31 days after turnout. Eggs per gram (EPG) differed between parasite exposure levels (P < 0.0001), with mean EPG remaining low in LP throughout the experiment. Similarly, an increase in SPC was observed (P < 0.0001), but only in HP where it peaked at day 56. In contrast, no difference in BW gain (BWG) (P = 0.78) between HP and LP was observed. In conclusion, this study shows that behavioural measurements monitored with sensors were affected even at low infection levels not affecting BWG. These combined results demonstrate the potential of automated behavioural recordings as a tool for detection of subclinical parasitism.

Selective flukicide treatment of non-lactating cows and the corresponding production impact of Fasciola hepatica in dairy herds in Sweden.

A control strategy against Fasciola hepatica infection based on selective treatment of non-lactating animals was evaluated in four Swedish dairy herds. The study was conducted over the course of two consecutive seasons in moderately to highly F. hepatica infected herds with robotic milking, where heifers and dry cows received an oral drench with albendazole (10 mg/kg) during three visits in January, February and March in both 2017 and 2018. This resulted in an anthelmintic coverage between 38 % and 58 % of the animals. Furthermore, on each visit, the infection status of all dewormed animals along with 15 randomly selected milking cows were monitored by detection of F. hepatica coproantigens. Individual milk samples were also collected quarterly from the whole herds for measurements of individual antibody levels against the parasite using milk ELISA. In addition, individual data on milk yield and quality were collected on a monthly basis between 2016 and 2018. To further study the impact of the infection on milk production, truly F. hepatica positive and negative cows in the first lactation were identified based on the results from coproantigen and milk ELISA assays. Total F. hepatica coproantigen prevalence in the herds varied between 28 % and 85 % in the first year, and between 27 % and 68 % in the second year of the study. We found that two years of treatments resulted in a significant decrease of coproantigen-positivity especially on the two most heavily infected farms. These results were confirmed by a similar drop in within-herd prevalences obtained by milk ELISA results. The infection had a significant negative impact on milk yields in untreated F. hepatica positive cows. No consistent long-term effect was observed at the herd level probably due to the influx of animals infected before puberty and/or adult animals that were re-infected at dry-off. This is the first study of the effects of F. hepatica infection on milk yield and quality in dairy herds in Sweden.

Effects of nematode parasitism on activity patterns in first-season grazing cattle.

We investigated the effects of gastrointestinal nematode (GIN) challenge on activity patterns in first season grazing (FSG) steers exposed to two different levels of Ostertagia ostertagi and Cooperia oncophora. At turn-out, experimental animals were allocated to one of two treatment groups grazing in different enclosures each with 32 animals. The first group (High) received 5000 third stage (L3) O. ostertagi (50%) and C. onchophora (50%) larvae; whereas the second group (Low) were dewormed monthly with 0.5 mg ivermectin (Noromectin®, Pour-on) per kg bodyweight. Activity patterns were monitored by fitting some animals in each group (High, n = 10; Low, n = 8) with leg mounted sensors (IceTag® 3D-accelerometers) during three two-week periods. In animals fitted with sensors body weight gain (BWG) was recorded every fortnight, whereas faecal and blood samples were collected every four weeks for nematode faecal egg count (FEC) and serum pepsinogen concentrations (SPC). Differences between the periods in daily (P = 0.046) and diurnal (P = 0.0502) activities were recorded between groups during the course of the study. A significant (P = 0.038) increase in the number of lying bouts was recorded in group High during the second period (days 74-86), which was correlated (r = 0.55, P = 0.018) to an increase in SPC ≈ 85 days after turn-out. BWG was reduced (P = 0.037) in group High compared to group Low, deviating from day 45. Strongyle nematode eggs were observed in both groups 29 days after turn-out, however the mean EPG remained low in group Low throughout the experiment. An increase in SPC was observed (P < 0.0038) in group High with levels peaking on day 58. In conclusion, our data supports that changes in activity patterns monitored with sensors could contribute to the identification of animals challenged with GIN, but also improve our understanding in the potential welfare impairments caused by such infections.

Effects of nematode parasitism on activity patterns in first-season grazing cattle.

We investigated the effects of gastrointestinal nematode (GIN) challenge on activity patterns in first season grazing (FSG) steers exposed to two different levels of Ostertagia ostertagi and Cooperia oncophora. At turn-out, experimental animals were allocated to one of two treatment groups grazing in different enclosures each with 32 animals. The first group (High) received 5000 third stage (L3) O. ostertagi (50%) and C. onchophora (50%) larvae; whereas the second group (Low) were dewormed monthly with 0.5 mg ivermectin (Noromectin®, Pour-on) per kg bodyweight. Activity patterns were monitored by fitting some animals in each group (High, n = 10; Low, n = 8) with leg mounted sensors (IceTag® 3D-accelerometers) during three two-week periods. In animals fitted with sensors body weight gain (BWG) was recorded every fortnight, whereas faecal and blood samples were collected every four weeks for nematode faecal egg count (FEC) and serum pepsinogen concentrations (SPC). Differences between the periods in daily (P = 0.046) and diurnal (P = 0.0502) activities were recorded between groups during the course of the study. A significant (P = 0.038) increase in the number of lying bouts was recorded in group High during the second period (days 74-86), which was correlated (r = 0.55, P = 0.018) to an increase in SPC ≈ 85 days after turn-out. BWG was reduced (P = 0.037) in group High compared to group Low, deviating from day 45. Strongyle nematode eggs were observed in both groups 29 days after turn-out, however the mean EPG remained low in group Low throughout the experiment. An increase in SPC was observed (P < 0.0038) in group High with levels peaking on day 58. In conclusion, our data supports that changes in activity patterns monitored with sensors could contribute to the identification of animals challenged with GIN, but also improve our understanding in the potential welfare impairments caused by such infections.

Genes regulating tight junctions and cell adhesion are altered in early experimental necrotizing enterocolitis.

BACKGROUND/PURPOSE: Necrotizing enterocolitis (NEC) represents one of the gravest complications in preterm infants and carries significant morbidity and mortality. Increased intestinal permeability may play an important role in the pathogenesis of NEC. In this study we investigated the genes regulating structural proteins such as tight junctions (TJ) and cell adhesion in a neonatal rat model of early NEC. METHODS: The studies were performed on Sprague-Dawley rat pups. Experimental NEC was induced using hypoxia/re-oxygenation treatment on day 1 after birth. Intestinal specimens from the ileum were obtained, mRNA was purified, and the transcriptome was analyzed using microarray. RESULTS: We found several TJ genes such as claudins 1, 8, 14, 15, and gap junction protein to be affected. Alterations in genes involved in the inflammatory response was confirmed, along with several genes regulating proteins used as biomarkers for NEC. CONCLUSION: This study indicates that tight junctions and cell adhesion may play a critical role in the pathogenesis of early experimental NEC. Better understanding of the pathogenesis of NEC may lead to novel strategies for the prevention and treatment of NEC.

Intraluminal intestinal microdialysis detects markers of hypoxia and cell damage in experimental necrotizing enterocolitis.

BACKGROUND/PURPOSE: Necrotizing enterocolitis (NEC) represents one of the gravest complications in premature infants and carries significant morbidity and mortality. There is a great need for improved diagnostic methods to reduce the severity and incidence of NEC. The aim of the study was to investigate if intraluminal microdialysis can detect intestinal ischemia in newborn rats with induced experimental NEC. METHODS: The studies were performed on 1-day-old Sprague-Dawley rat pups. Experimental NEC was induced using hypoxia/reoxygenation treatment. Microdialysis catheters were rectally inserted and placed in the rectosigmoid part of the colon. Microdialysate levels of glucose, lactate, pyruvate, and glycerol were measured. Intestinal specimens were collected at the end of the experiments for microscopic evaluation. RESULTS: Intraluminal microdialysis revealed signs of intestinal hypoxia and cellular damage, with a marked increase of lactate and glycerol. Microscopic evaluation confirmed intestinal damage in the NEC group. CONCLUSION: Intraluminal microdialysis can detect intestinal hypoxic stress and mucosal cell membrane decay in a rat model of NEC. Intestinal intraluminal microdialysis is easily accessible through the rectum and may be a useful noninvasive complement to other methods in the assessment of NEC.