Animals, Volume 14, Issue 7 (April-1 2024) – 151 articles

Laboratory animal facility managers must ensure that animal experiments can be carried out under optimal scientific conditions, that all legal requirements are met, and that animal welfare is maximized. Animal experimentation is stressful not only for the animals involved but also for the people who maintain these animals or carry out the experiments. Many of those involved find themselves in a constant conflict between scientific necessity, care, and harm. Under the term Culture of Care, procedures have been developed to reduce the burden of animal experimentation on the animals and the staff involved. The focus here is on what laboratory animal facility managers can do to improve the welfare of laboratory animals and the people working with them. Exemplary measures are the improvement of the housing conditions of laboratory animals, the introduction of uniform handling measures, clear and transparent structures via a quality management system, implementation of a no-blame culture of error (e.g., via Critical Incident Reporting System in Laboratory Animal Science [CIRS-LAS]), and open and respectful communication with all parties involved in animal experimentation, including the public and representatives of the authorities (public webpage, open house policy). The 6 Rs must be considered at all times: replacement, reduction, refinement, respect, responsibility, and reproducibility. We are writing this article from the perspective of laboratory animal facility managers in Germany. Full article

This research aimed to study the rostral part of the head of the six-banded armadillo, applying advanced imaging techniques such as CT. Furthermore, by combining the images obtained through this technique with anatomical cross-sections, an adequate description of the structures that constitute the rostral part of the head of this species is presented. This anatomical information could provide a valuable diagnostic tool for the clinical evaluation of different disorders in the six-banded armadillo’s nasal cavity and paranasal sinuses. Full article

Biosecurity practices aim to reduce the frequency of disease outbreaks in a farm, region, or country and play a pivotal role in fortifying the country’s pork industry against emerging threats, particularly foreign animal diseases (FADs). This article addresses the current biosecurity landscape of the US swine industry by summarizing the biosecurity practices reported by the producers through the United States Swine Health Improvement Plan (US SHIP) enrollment surveys, and it provides a general assessment of practices implemented. US SHIP is a voluntary, collaborative effort between industry, state, and federal entities regarding health certification programs for the swine industry. With 12,195 sites surveyed across 31 states, the study provides a comprehensive snapshot of current biosecurity practices. Key findings include variability by site types that have completed Secure Pork Supply plans, variability in outdoor access and presence of perimeter fencing, and diverse farm entry protocols for visitors. The data also reflect the industry’s response to the threat of FADs, exemplified by the implementation of the US SHIP in 2020. As the US SHIP program advances, these insights will guide industry stakeholders in refining biosecurity practices, fostering endemic re-emerging and FAD preparedness, and ensuring the sustainability of the swine industry in the face of evolving challenges. Full article

Domestic chicken farming has been promoted and spread in several Italian municipalities and worldwide as an aid to the self-consumption of domestically produced food. This study investigated the levels of four toxic elements (As, Cd, Hg, and Pb) in eggs from an ethical laying hen farm, comparing the element concentrations with those possibly present in supermarket eggs. A total of 201 eggs, 141 from the farm and produced by different hen genotypes, and 60 from the supermarket, were collected. The levels of the toxic elements were evaluated in the yolk, albumen, and eggshells of all eggs. The results show that the supermarket eggs’ yolk and albumen were more contaminated with lead, compared to the rural eggs. Contrarily, the mean content of arsenic was higher in the albumen and eggshells of the rural eggs, compared to the supermarket eggs. The cadmium content was below the LOQ (0.005 mg/kg) in all samples. The mercury content was below or around the LOQ in all rural eggs. Overall, the supermarket egg albumens were significantly more contaminated than the rural ones. No significant differences were found in quality parameters for both types of eggs. The toxic element values that were detected were in line with other studies in the literature. However, despite the concentrations found not representing a risk to the consumers’ health, the results of this study raise a potential food safety issue, and it would be desirable to set specific MRLs for eggs for consumers’ protection. Full article

Animal personality, which describes inter-individual differences and intra-individual consistency in behaviors across time and contexts, has been widely observed and has significance for both ecology and evolution. Morphological modifications, particularly during early life stages, may highly influence animal behavior in adulthood; thus, exploring this relationship can elucidate personality development throughout ontogeny. In this study, we reared juvenile crayfish (Procambarus clarkii) with different degrees of cheliped mutilation and explored their personality patterns, including exploration and aggression, when they reached sexual maturity. Male crayfish showed repeatability in exploration, and both sexes showed repeatability in aggression. We observed no significant correlation between the two behavioral traits, indicating the absence of behavioral syndromes. Moreover, exploration did not differ according to the type of mutilation, but crayfish with more intact chelipeds were more aggressive, and males were more aggressive than females. These results indicate that cheliped mutilation may modify the average levels of personality traits associated with competition or self-defense. Our study provides insights into how morphological modifications may shape animal personalities in adulthood. Full article

This comprehensive study on the Andalusian Black cattle breed reveals a substantial population decline, with the average herd size decreasing significantly from 305.54 to 88.28 animals per herd. This decline is primarily attributed to agricultural changes and the introduction of foreign meat-focused breeds. The male-to-female ratio shift is noteworthy, with more cows than bulls, impacting selection intensity for both genders. Inbreeding levels, though relatively low historically (5.94%) and currently (7.23%), raise concerns as 37.08% historically and 48.82% currently of the animals exhibit inbreeding. Positive assortative mating is evident, reflected by the increasing non-random mating coefficient (α). Key ancestors play a crucial role in shaping genetic diversity, with one ancestor significantly influencing the current genetic pool and the top 10 ancestors contributing substantially. Breed maintains a conservation index of 2.75, indicating relatively high genetic diversity. Recent conservation efforts have led to an increase in registered animals. The Cañadas Reales, historical transhumance routes, may have contributed to genetic connections among provinces. Challenges include the historical bottleneck, demographic changes, and potential impacts from reproductive practices. The Andalusian Black breed’s conservation necessitates ongoing efforts in genealogical registration, targeted breeding programs, and collaborative initiatives to address the observed demographic shifts and ensure sustainable genetic diversity. Full article

Colostrum contains macro- and micronutrients necessary to meet the nutritional and energy requirements of the neonatal calf, bioactive components that intervene in several physiological aspects, and cells and microorganisms that modulate the calf’s immune system and gut microbiome. Colostrum is sometimes mistaken as transition milk, which, although more nutritive than whole milk, has a distinct biochemical composition. Furthermore, most research about colostrum quality and colostrum management focuses on the transfer of maternal IgG to the newborn calf. The remaining components of colostrum and transition milk have not received the same attention, despite their importance to the newborn animal. In this narrative review, a large body of literature on the components of bovine colostrum was reviewed. The variability of these components was summarized, emphasizing specific components that warrant deeper exploration. In addition, the effects of each component present in colostrum and transition milk on several key physiological aspects of the newborn calf are discussed. Full article

Biowaste from slaughterhouses can be recovered to benefit food security and reduce contamination potential. More than 3 billion heads of livestock are consumed worldwide, which will increase by 17% by 2028, generating more biowaste, increasing infectious agents, and causing economic losses due to circular economy principles not being applied. This work evaluated the nutritional quality of four types of biowaste from bovine slaughter which were transformed into a meal for guinea pigs (rumen content (RCM), ears (EaM), blood (BM), and cheeks (CM)) according to their chemical composition, digestible components, energy contribution, and voluntary consumption. For the animal model, adult male guinea pigs were arranged in metabolic cages for feces collection without urinary contamination. Nine guinea pigs were used in each digestibility test. First, a direct digestibility test was conducted using a meal of barley as a reference diet (RD), the indigestibility coefficient of which allowed for the estimation of the digestibility of biowaste meals through indirect calculations; for this, diets composed of 80% of the RD and 20% of the corresponding biowaste meals were evaluated. The difference method was suitable for determining the digestibility of beef biowaste using the indigestibility coefficients of the reference diet to calculate the digestibility of ingredients which could not be offered as 100% of the meal but were incorporated as 20%. The digestible protein and metabolizable energy contents of RCM, EaM, BM, and CM were 10.2% and 2853 kcal/kg, 44.5% and 3325 kcal/kg, 70.7% and 2583 kcal/kg, and 80.8% and 3386 kcal/kg, respectively. The CM and BM feeds had the highest contributions of digestible protein due to their higher nitrogen content, and the CM and EaM feeds had the highest ME contents due to their higher fat contents. The biowaste meal consumption in descending order was CM > RCM > EaM > BM, which were consumed without problems. These results are indicative that these components can be part of guinea pigs’ diets, and it is recommended to continue studies into guinea pig growth and fattening diets with different levels of these biowaste meals. Full article

While paratuberculosis control has been studied for over a century, knowledge gaps still exist regarding the uptake and efficacy of control programmes. This narrative review aims to summarise studies on control programmes presented at the IDF ParaTB Fora in 2021 and 2022 and the International Colloquium on Paratuberculosis in 2022. Studies were grouped by topic as follows: successful control, field studies, education and extension, voluntary and compulsory control programmes, and surveillance. Various Map control programmes resulted in a decreasing animal and herd level Map prevalence. Long-term stakeholder commitment, stable funding, involvement of herd veterinarians and incentives for farmers to participate were shown to be pivotal for long-term success. Control measures focused on vertical and calf-to-calf transmission may improve Map control in infected herds. Easy-to-capture visualisation of surveillance test results to inform participants on the progress of Map control in their herds was developed. The probability of freedom from disease and estimated within-herd prevalence were identified as good candidates for categorisation of herds to support low-risk trade of cattle. Results of the surveillance schemes may inform genetic selection for resistance to Map infection. In conclusion, successful paratuberculosis control is feasible at both the herd and country level provided that crucial prerequisites are met. Full article

The Chinese goral (Naemorhedus griseus) is identified as a vulnerable species on the Red List of China’s Biodiversity and listed as a national second-class key protected wild animal in China. It is a representative flagship ungulate in Beijing. Its distribution range is fragmented and small populations are separated by dense infrastructures and tourism sites. Understanding its population status provides a foundation to plan effective conservation strategies. In this study, a population viability analysis was conducted with VORTEX (10.5.6.0) on a Chinese goral population in Beijing Yunmengshan Nature Reserve with the data collected by camera trapping and parameters referenced from other goral populations. The results show that this population will initially increase in the next 20 years and then decrease with a 32% probability of extinction risk. Supplementation with four adults, two females and two males, every 10 years would help minimize the extinction risk of this population. The results highlight the vital limiting factors for Chinese goral, including the initial population size, sex ratio at birth and mortality of infants (especially female infants). To improve the protection efficiency, detailed population parameters should be further acquired through continuous monitoring of this population. A thorough large-scale study should be carried out on other segregated goral populations in Beijing to facilitate the recovery of this endangered species. Full article

Ovulation and artificial insemination rates have been observed to decrease in sheep and cows when exposed to dietary phytoestrogens at concentrations greater than 25 mg/kg DM. A grazing trial was undertaken to investigate the effects of coumestrol and other key phytoestrogens on the superovulatory response, embryo numbers and quality in beef cows grazing legume pastures. A 7-week controlled grazing trial was conducted with legume and ryegrass pasture treatments, with cows exposed to legumes at two timed treatments, 4 and 7 weeks. Twenty Angus cows were subjected to a conventional estrus synchronization and superovulation protocol. Embryos were recovered via conventional uterine body flushing 7 days post artificial insemination (AI). Numerous phytoestrogens were identified in both pasture and plasma samples, including coumestrol and formononetin. Concentrations of phytoestrogens in the pasture ranged from 0.001 to 47.5 mg/kg DM and 0 to 2.6 ng/mL in plasma. Approximately 50% of cows produced viable embryos 7 days post AI. A significant interaction between the effect of treatment groups on the embryo stage was observed (p < 0.05). The results suggest that concentrations of >25 mg/kg DM of phytoestrogens less than 20 days preceding AI may negatively affect oocyte developmental competence, reduce progesterone production and thus contribute to early embryonic loss. Full article

The Mongolian racerunner, Eremias argus, is a small lizard endemic to Northeast Asia that can serve as an excellent model for investigating how geography and past climate change have jointly influenced the evolution of biodiversity in this region. To elucidate the processes underlying its diversification and demography, we reconstructed the range-wide phylogeographic pattern and evolutionary trajectory, using phylogenetic, population genetic, landscape genetic, Bayesian phylogeographic reconstruction and ecological niche modeling approaches. Phylogenetic analyses of the mtDNA cyt b gene revealed eight lineages that were unbounded by geographic region. The genetic structure of E. argus was mainly determined by geographic distance. Divergence dating indicated that E. argus and E. brenchleyi diverged during the Mid-Pliocene Warm Period. E. argus was estimated to have coalesced at~0.4351 Ma (Marine Isotope Stage 19). Bayesian phylogeographic diffusion analysis revealed out-of-Inner Mongolia and rapid colonization events from the end of the Last Interglacial to the Last Glacial Maximum, which is consistent with the expanded suitable range of the Last Glacial Maximum. Pre-Last Glacial Maximum growth of population is presented for most lineages of E. argus. The Glacial Maximum contraction model and the previous multiple glacial refugia hypotheses are rejected. This may be due to an increase in the amount of climatically favorable habitats in Northeast Asia. Furthermore, E. argus barbouri most likely represents an invalid taxon. The present study is the first to report a range-wide phylogeography of reptiles over such a large region in Northeast Asia. Our results make a significant contribution towards understanding the biogeography of the entire Northeast Asia. Full article

Positive reinforcement and training for health optimization are pivotal for successful studies with monkeys. Potential food inclination is important for studies on crab-eating macaques in laboratory environments, but evaluations remain scarce. We explored crab-eating macaques’ potential food inclination to establish a reward system for future behavioral assessments. Twelve male and three female monkeys underwent a food inclination assessment in which they were offered four food categories—fruits, vegetables, proteins, and nuts. The monkeys exhibited a higher inclination for plant-based foods, particularly fruits and vegetables, over animal-based proteins like chicken and tuna (p < 0.0001), with a notable inclination for nuts (eaten/provided = 100%). Additionally, the consistency of potential food inclination after repeated offerings was investigated, revealing a time-dependent increase in inclination for protein items. Food consumption ratios correlated positively with caloric intake (r = 0.59, p = 0.02), implying that individuals with a regular high caloric intake and increased body weight are more likely to accept food during positive reinforcement training. Our findings suggest fruits, vegetables, protein-rich foods, and nuts can help with health optimization. However, animal-based protein-rich foods initially had a low preference, which may increase over time. Our study can provide guidelines for positive reinforcement training and health optimization. Full article

This paper aims to identify Poria cocos polysaccharides (PCPs) as a potential feed additive used for swine production; thus, we explored the effects of different dietary inclusion levels of PCP on growth performance, immunity, and cecal microflora composition in weaned piglets. For this, a total of 120 28-day-old Duroc × Landrace × Yorkshire weaned piglets (8.51 ± 0.19 kg; 28 ± 1 days of age) were randomly allocated to five groups that were fed a basal diet supplemented with 0, 0.025%, 0.05%, 0.1%, and 0.2% PCP, respectively, for 42 days. The results indicated that the average daily gain (ADG) and gain/feed ratio were higher in the PCP treatment groups than in the control group, with a linear effect. The serum concentrations of IgG, IgA, IL-2, IFN-γ, the number of CD4⁺ T cells, and the CD4⁺-to-CD8⁺ T-cell ratio (CD4⁺/CD8⁺) were increased, while the levels of IL-6 and TNF-α were decreased in the PCP supplementation groups compared with those in the control group. Furthermore, the cytokine mRNA expression levels exhibited a similar trend in the spleen. PCP supplementation also reduced the abundance of Escherichia coli and Salmonella and enhanced that of Lactobacilli and Bifidobacteria in the cecum. In summary, dietary PCP inclusion exerted positive effects on the growth performance, immunity, and cecal microbiota of piglets and showed potential for use as a feed additive for improving the health of weaned piglets, with 0.1% being the optimal dosage. Full article

Subchondral cystic lesions (SCL) in the medial femoral condyle are a usual finding in Thoroughbred survey and auction repository radiographs. Several treatments with different outcomes have been studied over the years to improve soundness and racing prognosis. Our objective was to report the racing prognosis in Thoroughbred yearlings intended for racing that were diagnosed with SCL in the medial femoral condyle and were treated using four current and different techniques: intralesional injection of corticosteroids, SCL debridement through the joint with a drill bit, translesional cortical screw placement, and absorbable hydroxyapatite implant placement. Data from 182 Thoroughbred yearlings treated for SCL in the medial femoral condyle were collected from 2014 to 2020. Limb affected, age at surgery, sex, and radiographic measurements of the SCL were recorded. Auction price and racing performance were collected for treated horses and compared to 154 maternal siblings free of medial femoral condyle SCL. Analyses were conducted to assess if racing prognosis was affected by SCL size, to detect differences in auction price and selected flat racing outcome parameters between cases and controls, and to compare racing prognosis between the studied treatments. Mares and lesions located in the right stifle were significantly overrepresented. The auction price of treated horses was significantly lower than that of their siblings. Horses treated for SCL had significantly lower chances to start in a race than controls (59% vs. 74% respectively). Wider SCL negatively affected the chances to start at least in one race, and negatively affected the earnings made in the 2-year-olds’ racing year. Horses with SCL treated using a bioabsorbable implant had a significantly higher median in starts as 3-year-olds (seven starts) than horses that had the SCL debrided with a drill bit (three starts). In conclusion, Thoroughbred yearlings treated for a medial femoral condyle SCL had lower auction prices and decreased ability to start a race compared to siblings’ wider cysts had worse prognosis to start a race and might affect earnings as 2-year-olds; and horses treated with bioabsorbable composite implant placement had more starts as 3-year-olds than with other techniques. Full article

Cognitive enrichment is becoming more prevalent in professional marine mammal facilities. Research with dolphins has suggested that such enrichment provides more welfare benefits than enrichment that does not incorporate cognitive challenge. However, there is little research supporting the use of cognitive enrichment as a means to improve the welfare of sea lions. Recently, a novel form of technological cognitive enrichment, the Enclosure Video Enrichment (EVE) system, was introduced to a population of California sea lions at the US Navy’s Marine Mammal Program as a means to enhance welfare. Two of the initial focal animals introduced to EVE were selected based on their health history and the possible benefits of cognitive enrichment in improving health measures. To evaluate this, information regarding the animals’ consumption of their offered diet was compared to other animals in the population of similar age and the absence of a similar health history. Subsequently, the total diet consumption of the targeted animals was evaluated in the two years prior to the introduction to EVE and compared to the total diet consumption during the initial 2 years of regular EVE sessions. There was a significant decrease in the number of days in which the sea lions did not consume their entire offered diet in the two years after implementing regular EVE sessions, an increase in participation and performance of voluntary husbandry behaviors, and a reduction in the number of days animals were clinically ill. This study provides evidence of cognitive enrichment as a management tool to improve animal health and welfare as well as performance in training sessions. Full article

Small ruminant lentiviruses are a group of viruses infecting goat and sheep worldwide. These viruses exhibit an extraordinary degree of genetic and antigenic variability that severely influence in vivo and in vitro features, as well as diagnostic test results. Small ruminant farming is the most important animal farming business in Greece, with a high impact on the Greek primary economy. Although SRLV infection and its impact on animal production are well established in the country, little is known about the circulating SRLV strains and their prevalence. The aim of this study was to characterize SRLVs circulating in Greece with a combined serological and molecular approach, using the bulk milk matrix collected from 60 farms in different municipalities. This study allowed us to estimate a seroprevalence of around 52% at the herd level. The B1, B2 and A3 subtypes and a novel A viral cluster were identified. Moreover, the amplicon sequencing method allowed us to identify more than one viral subtype in a sample. These results again confirm the high variability of these viruses and highlight the importance of the constant monitoring of viral evolution, in particular in antigens of diagnostic interest. Full article

The corpus luteum is a temporary endocrine gland formed in the ovary after ovulation, and it plays a critical role in animal reproductive processes. Tumors rely on the development of an adequate blood supply to ensure the delivery of nutrients and oxygen and the removal of waste products. While angiogenesis occurs in various physiological and pathological contexts, the corpus luteum and tumors share similarities in terms of the signaling pathways that promote angiogenesis. In the corpus luteum and tumors, apoptosis plays a crucial role in controlling cell numbers and ensuring proper tissue development and function. Interestingly, there are similarities between the apoptotic-regulated signaling pathways involved in apoptosis in the corpus luteum and tumors. However, the regulation of apoptosis in both can differ due to their distinct physiological and pathological characteristics. Thus, we reviewed the biological events of the corpus luteum and tumors in similar microenvironments of angiogenesis and apoptosis. Full article

The contamination of aquatic ecosystems by the heavy metal copper (Cu) is an important environmental issue and poses significant risks to the physiological functions of aquatic organisms. Macrobrachium rosenbergii is one of the most important freshwater-cultured prawns in the world. The hepatopancreas of crustaceans is a key organ for immune defense, heavy metal accumulation, and detoxification, playing a pivotal role in toxicological research. However, research on the molecular response of the hepatopancreas in M. rosenbergii to Cu exposure is still lacking. In this study, the transcriptomic response in the hepatopancreas of M. rosenbergii was studied after Cu exposure for 3 and 48 h. Compared with the control group, 11,164 (7288 up-regulated and 3876 down-regulated genes) and 10,937 (6630 up-regulated and 4307 down-regulated genes) differentially expressed genes (DEGs) were identified after 3 and 48 h exposure, respectively. Most of these DEGs were up-regulated, implying that gene expressions were largely induced by Cu. Functional enrichment analysis of these DEGs revealed that immunity, copper homeostasis, detoxification, DNA damage repair, and apoptosis were differentially regulated by Cu. Seven genes involved in immunity, detoxification, and metabolism were selected for validation by qRT-PCR, and the results confirmed the reliability of RNA-Seq. All these findings suggest that M. rosenbergii attempts to resist the toxicity of Cu by up-regulating the expression of genes related to immunity, metabolism, and detoxification. However, with the excessive accumulation of reactive oxygen species (ROS), the antioxidant enzyme system was destroyed. As a result, DNA damage repair and the cellular stress response were inhibited, thereby exacerbating cell damage. In order to maintain the normal function of the hepatopancreas, M. rosenbergii removes damaged cells by activating the apoptosis mechanism. Our study not only facilitates an understanding of the molecular response mechanisms of M. rosenbergii underlying Cu toxicity effects but also helps us to identify potential biomarkers associated with the stress response in other crustaceans. Full article

The growth of the broiler industry has been accompanied with concerns over the environmental and social impacts on intensive production systems, as well as the welfare of the animals themselves. As a result, since the 2000s, there has been growing interest in alternative production systems that improve animal welfare and sustainability. In this context, it is important to prioritize the welfare of broilers in commercial production systems and to use reliable welfare indicators to provide consumers with information about the welfare of the animals they consume. Resource-based measures (RBM) are based on assessing the resources available to the birds in terms of their housing, environment, and management practices, such as stocking density, litter quality, lighting and air quality, etc. Outcome-based measures (OBM), also known as animal-based measures, focus on assessing the actual welfare outcomes for the birds, such as footpad dermatitis, hock burn, contamination or damage to feathers, gait score, mortality, etc. These OBM-based measures are one of the more direct indicators of welfare and can help identify any welfare issues. The present review highlighted the factors that affect animal welfare indicators focused on OBMs which can be used in the commercial broiler farms. Full article

The porcine bulbourethral glands produce a gel-type secretion. Although the role of these contributions to reproductive success remains murky, the bulbourethral glands are major accessory sex glands in this species. Isometric growth in the early neonatal interval is followed by allometric growth in the late juvenile interval (6 to 11 weeks of age), while circulating endogenous steroids are low. The rate of allometric growth increases during the peripuberal interval (16 to 20 weeks of age) when systemic testosterone is relatively high. Gene expression for androgen receptor (AR) and for the steroid 5 alpha-reductase 2 (SRD5A2) enzyme that synthesizes the more potent androgen dihydrotestosterone from its precursor was evaluated by qPCR analyses of bulbourethral gland tissue. Tissues were collected from control boars (2 weeks to 40 weeks of age) and from littermates of these boars treated with letrozole to suppress endogenous estrogen synthesis. Gene expression for these two key proteins in androgen signaling was quite low during the initial allometric growth in the late juvenile and prepuberal intervals, suggesting that this initial growth was not primarily stimulated by androgens. These observations are consistent with a more direct estrogen-mediated inhibition of growth via GPER previously proposed, with the sensitivity extending into the late juvenile interval when estrogens as well as androgens are normally relatively low. Full article

Due to the fencing of the Przewalski’s gazelle (Procapra przewalskii), the microcytic anemia incidence rate continues to increase. The primary pathological symptoms include emaciation, anemia, pica, inappetence, and dyskinesia. To investigate the cause of microcytic anemia ailment in the Przewalski’s gazelle, the Upper Buha River Area with an excessive incidence was chosen as the experimental pasture, and the Bird Island Area without microcytic anemia disease was chosen as the control field. Then, the mineral contents in the soil, forage, blood, and liver, as well as the blood routine parameters and biochemical indexes were measured. The findings showed that the experimental pasture had much lower Se content in the soil and forage than the control field (p < 0.01), while the impacted pasture had significantly higher S content in the forage. The damaged gazelles had considerably lower Se and Cu contents and higher S content in the blood and liver than the healthy gazelles (p < 0.01). The presences of Hb, HCT, MCV, and MCH were significantly decreased compared to those in healthy gazelles (p < 0.01). The experimental group had a significantly lower level of GSH-Px activity in their serums compared to the control group (p < 0.01). In the treatment experiment, ten gazelles from the affected pasture were orally administered CuSO₄, 6 g/animal once every 10 days for two consecutive times, and all gazelles were successfully cured. Therefore, it is possible that low Se content in the soil induced an increase in the absorption of S content by forage, leading to the deficiency of secondary Cu in the Przewalski’s gazelles, resulting in microcytic anemia. Full article

Machine learning algorithms have been applied to various animal husbandry and veterinary-related problems; however, its use in Johne’s disease diagnosis and control is still in its infancy. The following proof-of-concept study explores the application of tree-based (decision trees and random forest) algorithms to analyze repeat milk testing data from 1197 Canadian dairy cows and the algorithms’ ability to predict future Johne’s test results. The random forest models using milk component testing results alongside past Johne’s results demonstrated a good predictive performance for a future Johne’s ELISA result with a dichotomous outcome (positive vs. negative). The final random forest model yielded a kappa of 0.626, a roc AUC of 0.915, a sensitivity of 72%, and a specificity of 98%. The positive predictive and negative predictive values were 0.81 and 0.97, respectively. The decision tree models provided an interpretable alternative to the random forest algorithms with a slight decrease in model sensitivity. The results of this research suggest a promising avenue for future targeted Johne’s testing schemes. Further research is needed to validate these techniques in real-world settings and explore their incorporation in prevention and control programs. Full article

Movements in cattle after captive bolt stunning cause problems in the slaughter process and lead to uncertainties in assessing stunning effectiveness. The objective of this study was to categorize and quantify these movements and determine animal- and process-related impact factors, as well as connections to stunning effectiveness and shooting position. In total 2911 cows, heifers, and bulls (dairy, beef, and crossbreeds) were examined (mean age 3.02 years). Movements from landing until at least four minutes after sticking were recorded by action cams (Apeman^® A100). Nine movement categories were defined (“kicking hind limb”, “twitching”, “bending and stretching hind limb”, “lifting and bending forelimb”, “body arching laterally”, “body arching ventrally”, and “arching backwards”). According to the movement severity, a score was assigned to each category. The scores were summed, either for certain process intervals, e.g., LANDING (ejection from the stunning box), HOISTING, or STICKING, or for the total time between LANDING and end of the FOURTH MINUTE OF BLEEDING (sum score). Statistical analysis (ANOVA) was performed on the scores. Only 6.6% of cattle showed no movement. Most movements occurred during STICKING and FIRST MINUTE OF BLEEDING, occurring rarely up to 8 min after sticking. While cows moved most at LANDING, bulls and heifers moved more if all process intervals were considered. The sum score was highest in German Angus, Charolais, and Limousin and lowest in Brown Swiss and Simmental. The score at LANDING was highest in German Angus and Black Holstein. The use of pneumatic stunners and an increase in bolt-exit length significantly reduced movements. No impact of stunning effectiveness on movements was found, but only 19 cattle showed reduced effectiveness. Full article

Guinea pigs (Cavia porcellus) are used for interactive activities in zoos; therefore, it is important to investigate their welfare. This study aimed to evaluate the validity of measuring the guinea pigs’ body temperature of guinea pigs through the ear canal and investigate the relationship among changes in the expression of negative behavior, changes in body temperature, and changes in salivary cortisol concentration, and examine the effects of different interactive activities. In the normal interactive activities performed at the site, the decreased body temperature of pigs was observed over time. In contrast, increased body temperature was observed in excessive interactive activities, which are not recommended. Among the negative behaviors, “Head turning” and “Locomotion” increased significantly in excessive interactions compared to normal interactions, but “Head tossing” decreased significantly over time in both types of interactions. “Freezing” was observed only in excessive interactions. Salivary cortisol concentration increased significantly for all activities. Investigating the relationship between the individual expression of negative behavior and changes in body temperature and changes cortisol level made it possible to uncover the potential for inferring an animal’s physiological state. Combining ear temperature monitoring and behavioral observation during zoo interaction activities is recommended as an ethical and scientifically supported practice. Full article

In the pig farming industry, it is recommended to avoid groups when treating individuals to reduce adverse reactions in the group. However, can this eliminate the adverse effects effectively? Piglets were assigned to the Rewarding Group (RG), the Punishing Group (PG), and the Paired Control Group (PCG). There were six replicates in each group, with two paired piglets per replicate. One piglet of the RG and PG was randomly selected as the Treated pig (TP), treated with food rewards or electric shock, and the other as the Naive pig (NP). The NPs in the RG and PG were unaware of the treatment process, and piglets in the PCG were not treated. The behavior and heart rate changes of all piglets were recorded. Compared to the RG, the NPs in the PG showed longer proximity but less contact behavior, and the TPs in the PG showed more freezing behavior. The percentage change in heart rate of the NPs was synchronized with the TPs. This shows that after sensory avoidance, the untreated pigs could also feel the emotions of their peers and their emotional state was affected by their peers, and the negative emotions in the pigs lasted longer than the positive emotions. The avoidance process does not prevent the transfer of negative emotions to peers via emotional contagion from the stimulated pig. Full article

Although direct behavioural observations are widely used, they are time-consuming, prone to error, require knowledge of the observed species, and depend on intra/inter-observer consistency. As a result, they pose challenges to the reliability and repeatability of studies. Automated video analysis is becoming popular for behavioural observations. Sleep is a biological metric that has the potential to become a reliable broad-spectrum metric that can indicate the quality of life and understanding sleep patterns can contribute to identifying and addressing potential welfare concerns, such as stress, discomfort, or health issues, thus promoting the overall welfare of animals; however, due to the laborious process of quantifying sleep patterns, it has been overlooked in animal welfare research. This study presents a system comparing convolutional neural networks (CNNs) with direct behavioural observation methods for the same data to detect and quantify dogs’ sleeping patterns. A total of 13,688 videos were used to develop and train the model to quantify sleep duration and sleep fragmentation in dogs. To evaluate its similarity to the direct behavioural observations made by a single human observer, 6000 previously unseen frames were used. The system successfully classified 5430 frames, scoring a similarity rate of 89% when compared to the manually recorded observations. There was no significant difference in the percentage of time observed between the system and the human observer (p > 0.05). However, a significant difference was found in total sleep time recorded, where the automated system captured more hours than the observer (p < 0.05). This highlights the potential of using a CNN-based system to study animal welfare and behaviour research. Full article

As a reactive species, the red deer is sensitive to both negative exogenous and endogenous stimuli. An intensive hunting period may have a particularly negative impact on game animals. The aim of this study was to determine the plasma cortisol level and biochemical parameters in 25 wild red deer (Cervus elaphus) harvested during stalking hunts in correlation with the sex and age of the animals. The mean cortisol concentrations in the stags and hinds analyzed in this study were similar (20.2 and 21.5 ng/mL, respectively). Higher HDL cholesterol values were found in the blood of the hinds than in stags (p < 0.05). Similarly, the mean levels of LDL cholesterol, lactate dehydrogenase, and alanine aminotransferase were higher by 21%, 16%, and 42%, respectively, in the blood of the hinds. In contrast, the levels of alkaline phosphatase, bilirubin, and aspartate aminotransferase were higher in the stags (by 30%, 49%, and 36%, respectively). There was a negative correlation of the cortisol concentration with urea and bilirubin and a positive correlation between cortisol and aspartate aminotransferase in the stags (p < 0.05). In turn, a negative correlation was found between the cortisol and urea levels in the hinds (p < 0.05). In summary, the stress caused by stalking hunts and the characteristic behavior of red deer during the mating season had an impact on chosen biochemical parameters. The increased concentration of cortisol resulted in a decrease in the carcass mass, which may lead to the deterioration of the physical condition of animals on hunting grounds. Full article

Here, we document the stranding of a striped dolphin Stenella coeruleoalba (Meyen, 1833) (Mammalia: Delphinidae), which was found dead in Maltese waters in July 2020. The stranded dolphin exhibited a severe infestation of the mesoparasitic copepod, Pennella balaenoptera Koren and Danielssen, 1877 (Copepoda: Pennelidae). Parasites of this genus represent the largest known mesoparasites to infest cetaceans. Under normal circumstances, cetaceans may have a few P. balaenoptera individuals attached to them, but cetaceans with compromised health are more prone to heavy infestations. The identification of the parasite was accomplished through morphological and genetic analyses. This incident highlights the significance of monitoring mesoparasitic infestations, offering valuable insights into the health of cetacean populations and emphasizing the potential implications for conservation efforts in the region. Full article