Animals, Volume 14, Issue 8 (April-2 2024) – 117 articles

The development of realistic dummies for training the distal phalanx amputation (DPA) technique in mouse pups is a promising alternative to reduce and replace animals in training for research and teaching. To test this, we obtained micro-CT data from postnatal day-five mouse pups, meticulously segmented them, and converted them into a 3D mesh format suitable for 3D printing. Once the dummy was printed, it was evaluated during actual training courses in two different groups: in the first group, users received no dummies to train the DPA, and in the second group, users were trained with three dummies. To assess the effectiveness of the dummy, we conducted a survey followed by an expert veterinarian evaluation. Our results showed that DPA is a complex procedure, and it is commonly poorly performed. When implementing the dummies, users who were not provided with dummies to practice only had an 8.3% success rate in DPA, while users provided with three dummies had a 45.5% success rate, respectively. Despite additional research being needed, our dummy offered improved practical training by providing a safe and effective alternative in line with ethical considerations while demonstrating the feasibility of using 3D printing technology to promote the 3Rs in experimental research. Full article

This examined the effects of Lonicera japonica extract (LJE) with different chlorogenic acid (CGA) contents on lactation performance, antioxidant status and immune function and rumen fermentation in heat-stressed high-yielding dairy cows. In total, 45 healthy Chinese Holstein high-yielding dairy cows, all with similar milk yield, parity, and days in milk were randomly allocated to 3 groups: (1) the control group (CON) without LJE; (2) the LJE-10% CGA group, receiving 35 g/(d·head) of LJE-10% CGA, and (3) the LJE-20% CGA group, receiving 17.5 g/(d·head) of LJE-20% CGA. The results showed that the addition of LJE significantly reduced RT, and enhanced DMI, milk yield, milk composition, and improved rumen fermentation in high-yielding dairy cows experiencing heat stress. Through the analysis of the serum biochemical, antioxidant, and immune indicators, we observed a reduction in CREA levels and increased antioxidant and immune function. In this study, while maintaining consistent CGA content, the effects of addition from both types of LJE are similar. In conclusion, the addition of LJE at a level of 4.1 g CGA/(d·head) effectively relieved heat stress and improved the lactation performance of dairy cows, with CGA serving as the effective ingredient responsible for its anti-heat stress properties. Full article

The objective of this dairy farm study was to investigate the preweaning health, performance, immunity, and enteric pathogen shedding in calves supplemented with colostrum during five days after birth compared to calves not supplemented with colostrum. The colostrum supplementation was the previously frozen colostrum added to the milk replacer from day 2–5 given to 39 calves, and 36 Control calves received a milk-derived supplement. There was no significant difference in preweaning weight gain between the treatment groups. Serum samples collected on days 2, 7, 14, and 21 indicated that total and antigen-specific IgG levels against rotavirus, coronavirus, and E. coli F5 were not significantly different between the treatment groups. Fecal samples taken on days 7, 14, and 21 were all negative for coronavirus and E. coli F5, whereas there were low levels of Cryptosporidia and a trend for low levels of rotavirus on day 14 in colostrum-supplemented compared to Control calves. Respiratory clinical signs, depressed attitude and body temperature tended to be reduced in colostrum-supplemented compared to Control calves. This study shows that, even in calves with good colostrum status and high plane of nutrition, there can be benefits of post-closure colostrum supplementation including reduced Cryptosporidia and rotavirus shedding and reduced respiratory disease. Full article

Mammary gland tumors represent the most frequently diagnosed malignant neoplasm in intact female dogs, and surgical removal represents the current gold standard treatment. To promote wound healing and prevent possible bacterial contamination, perioperative antimicrobials are commonly used in clinical practice, even though there are no publications establishing guidelines for the use of such drugs in canine mastectomy. The aim of the present study was to evaluate the ameliorative effect of fluorescent light energy on the quality of the healing process after mastectomy surgery in female dogs, in the absence of perioperative antimicrobial administration. Nine female dogs received a multiple-gland mastectomy due to gland tumors and received FLE application immediately after surgery and then five days after. The surgical incisions were evaluated by a blind investigator over time using the Modified Hollander Cosmesis and Modified Draize Wound Healing Score systems. Statistical analysis revealed a significant ameliorative effect of FLE in the control of step-off borders, contour irregularities, and excessive distortion. In addition, erythema, edema, and serous discharge were lower for those wounds managed with FLE. These results underscore the advantageous impact of FLE on the healing of post-mastectomy wounds in female dogs, offering the dual benefits of reducing potential infection risks and lessening the home care burden for pet owners. Full article

The study objectives were to understand the practices and perceptions of sheep farmers and district veterinarians (DVs) towards sheep health management and, the impact of disease at the farm level in addition to the availability, accessibility, and use of veterinary services. Data were collected using question-based surveys, distributed online and in-person to sheep farmers (45 respondents) and DVs (25 respondents). Most farmers were male, ≥51 years old, who placed a high priority on the health and welfare of their sheep. For disease prevention most farmers vaccinated their sheep (91%) and 86.7% had a farm biosecurity plan, although its components and their application varied, e.g., the isolation of new or sick sheep. Fencing costs were most frequently identified (70.5% respondents) as a financial concern for sheep farmers. Their most common sources of information about disease control and prevention were DVs (66.7%), private veterinarians (60.0%), the internet (42.9%), rural suppliers (35.7%), and farmers/neighbours (28.6%). Fifty-eight percent of farmers reported a long distance from veterinarian services. Farmers preferred to receive information via email (77.8%), whereas 56% of DVs preferred to share information via phone call. This discrepancy presents an opportunity to align these mechanisms more closely for effective dissemination of information and increased producer engagement. Full article

Household buffalo dairy farming is gaining popularity nowadays in Bangladesh because of the outstanding food value of buffalo milk as well as the lower production cost of buffalo compared to cattle. An initiative has recently been taken for the genetic improvement of indigenous dairy buffaloes. The present study was carried out to determine the influence of some environmental factors like age, parity, season of calving, calving interval, dry period on the lactation yield, and lactation curve of indigenous dairy buffaloes of Bangladesh. A total of 384 indigenous dairy buffaloes from the 3rd and 4th parity of seven herds under two different agroecological zones covering four seasons were selected and ear tagged for individual buffalo milk recording. A milk yield of 300 days (MY300d) was calculated following the International Committee for Animal Recording (ICAR) and the data were evaluated using the generalized linear model (GLM). In production traits, the mean of calculated lactation period (CLP), calculated lactation yield (CLY), and milk yield of 300 days (MY300d) of the overall population were 267.28 days, 749.36 kg, and 766.92 kg, respectively, whereas calving interval (CI) and dry period (DP) as reproductive traits were 453.06 days and 185.78 days, respectively. The season of calving, age of buffalo cows, population or herd, agroecological zone, calving interval, and dry period had significant effects on production traits (p < 0.05 to p < 0.001). The season of calving, level of milk production of 300 days, population, and agroecological zone significantly affected the reproduction traits (p < 0.01 to p < 0.001). Parity was found to be non-significant for both types of traits. The average peak yield of test day (TD) milk production was highest at TD4 (4.47 kg, 98th day of lactation). The average MY300d of milk production was the highest in the Lalpur buffalo population (1076.13 kg) and the lowest in the buffalo population of Bhola (592.44 kg). The correlations between milk production traits (CLP, CLY, and MY-300d) and reproduction traits (CI and DP) were highly significant (p < 0.01 to p < 0.001). Positive and high correlation was found within milk traits and reproduction traits, but correlation was negative between milk traits and reproduction traits. Therefore, these non-genetic factors should be considered in the future for any genetic improvement program for indigenous dairy buffaloes in Bangladesh. Full article

Equine Gastric Ulcers Syndrome (EGUS) is a worldwide disease present in equids of different breeds, activity levels, and age groups. It is divided into two different illnesses: Equine Squamous Gastric Disease (ESGD) affecting the squamous mucosa and Equine Glandular Gastric Disease (EGGD) affecting the glandular mucosa. The historical horserace is a traditional competition that is common in Italy. They can be divided into two different types: speed races and jousting tournaments. Anglo-Arabians and Thoroughbreds are used for the two competitions with training and management systems similar to those used in races performed on a racetrack. The aim of this study was to determine the prevalence of ESGD and EGGD in horses used for historical horseracing and evaluate the differences in the presence of the diseases in animals of the two different breeds used for the two types of competition. A cohort of 73 horses was enrolled in this study, which were stabled in 10 training facilities and performed two jousting tournaments and one speed race. An ESGD at least of grade 2 was found in 88% of horses with all degrees of severity seen; EGGD was diagnosed in 45% of animals. In this cohort of horses, the presence and severity of ESGD and EGGD are similar to that in reports in racehorses performing on racetracks. Anglo-Arabians used for speed races are more affected by ESGD and EGGD, which is probably because they are involved in a higher number of races and travel more during the year compared to Thoroughbreds used for jousting competitions. Full article

Cashmere goats are valuable genetic resources which are famous worldwide for their high-quality fiber. Runs of homozygosity (ROHs) have been identified as an efficient tool to assess inbreeding level and identify related genes under selection. However, there is limited research on ROHs in cashmere goats. Therefore, we investigated the ROH pattern, assessed genomic inbreeding levels and examined the candidate genes associated with the cashmere trait using whole-genome resequencing data from 123 goats. Herein, the Inner Mongolia cashmere goat presented the lowest inbreeding coefficient of 0.0263. In total, we identified 57,224 ROHs. Seventy-four ROH islands containing 50 genes were detected. Certain identified genes were related to meat, fiber and milk production (FGF1, PTPRM, RERE, GRID2, RARA); fertility (BIRC6, ECE2, CDH23, PAK1); disease or cold resistance and adaptability (PDCD1LG2, SVIL, PRDM16, RFX4, SH3BP2); and body size and growth (TMEM63C, SYN3, SDC1, STRBP, SMG6). 135 consensus ROHs were identified, and we found candidate genes (FGF5, DVL3, NRAS, KIT) were associated with fiber length or color. These findings enhance our comprehension of inbreeding levels in cashmere goats and the genetic foundations of traits influenced by selective breeding. This research contributes significantly to the future breeding, reservation and use of cashmere goats and other goat breeds. Full article

The aim of the present trial was to evaluate the growth performance of buffalo calves fed on diets characterized by different forage/concentrate ratios, with or without Saccharomyces cerevisiae supplementation (CBS 493.94, Yea-Sacc^®). Twenty-four male buffalo calves (mean age of 145.1 ± 16.1 days; mean weight of 108.0 ± 18.7 kg) were assigned randomly to 4 groups, homogeneous in age, that were fed four different diets: diet 1, F:C ratio 50:50; diet 2, F:C ratio 30:70; diet 3, F:C ratio 50:50 + Yea-Sacc^®; and diet 4, F:C ratio 30:70 + Yea-Sacc^®. Buffalo calves were individually weighted before the start of the experiment and the data were used as a covariate, being taken monthly until the end of the trial. Dry matter intake (DMI), daily weight gain (DWG) and feed conversion ratio (FCR) were calculated. The differences in diets composition significantly (p < 0.01) affected all these parameters. In particular, the animals fed diet 1 and diet 3 showed higher values of DWG (0.91 and 0.88 g/d vs. 0.68 and 0.66 for group 2 and 4) and DMI (5.8 and 5.3 kg/d, respectively) compared to the other groups (4.3 and 4.4 kg/d for group 2 and 4), as well as a higher final body weight (370.5 and 334.1 kg for group 1 and 3 vs. 272.8 and 273.1 kg of group 2 and 4, respectively). Indeed, the supplementation with Yea-Sacc^® at the dosage of 1 × 10^E8 did not affect buffaloes’ growth performance. Full article

Equine melanomas are a common neoplasm in gray horses. However, scientific knowledge about their progression over time is quite scarce. Some owners and veterinarians still believe that early intervention is not necessary, stating that tumors evolve very slowly and intervention could worsen the animal’s condition. This work aims to identify clinical and histological differences that may exist between equine melanomas with different excision intervals (time between tumor detection and surgical excision). A total of 42 tumors (13 benign and 29 malignant) from 34 horses were included in this study. There was a statistically significant association between excision interval and tumor size (p = 0.038), with tumors excised later being significantly larger than the ones excised sooner. The excision interval was also statistically associated with the number of tumors (p = 0.011), since the horses that carried a tumor for longer seemed to be prone to have multiple tumors. Furthermore, there was an association between excision interval and malignancy (p = 0.035), with tumor excised later being fives times more likely to be malignant. This study provides evidence of delayed excision’s effect on the progression of equine melanomas. Additionally, it reinforces the importance of the early excision of these tumors. Full article

Growth-retarded yaks are of a high proportion on the Tibetan plateau and reduce the economic income of farmers. Our previous studies discovered a maldevelopment in the ruminal epithelium of growth-retarded yaks, but the molecular mechanisms are still unclear. This study aimed to reveal how the proteomic profile in the ruminal epithelium contributed to the growth retardation of yaks. The proteome of the ruminal epithelium was detected using a high-resolution mass spectrometer. There were 52 proteins significantly differently expressed between the ruminal epithelium of growth-retarded yaks and growth-normal yaks, with 32 downregulated and 20 upregulated in growth-retarded yaks. Functional analysis showed the differently expressed proteins involved in the synthesis and degradation of ketone bodies (p = 0.012), propanoate metabolism (p = 0.018), pyruvate metabolism (p = 0.020), and mineral absorption (p = 0.024). The protein expressions of SLC26A3 and FTH1, enriched in the mineral absorption, were significantly downregulated in growth-retarded yaks. The key enzymes ACAT2 and HMGCS2 enriched in ketone bodies synthesis and key enzyme PCCA enriched in propanoate metabolism had lower protein expressions in the ruminal epithelium of growth-retarded yaks. The ATP concentration and relative mitochondrial DNA copy number in the ruminal epithelium of growth-normal yaks were dramatically higher than those of growth-retarded yaks (p < 0.05). The activities of citrate synthase (CS), the α-ketoglutarate dehydrogenase complex (α-KGDHC), isocitrate dehydrogenase (ICD) in the tricarboxylic acid cycle (TCA), and the mitochondrial respiratory chain complex (MRCC) were significantly decreased in ruminal epithelium of growth-retarded yaks compared to growth-normal yaks (p < 0.05). The mRNA expressions of COQ9, COX4, and LDHA, which are the encoding genes in MRCC I, IV and anaerobic respiration, were also significantly decreased in the ruminal epithelium of growth-retarded yaks (p < 0.05). Correlation analysis revealed that the average daily gain (ADG) was significantly positively correlated to the relative mitochondrial DNA copy number (p < 0.01, r = 0.772) and ATP concentration (p < 0.01, r = 0.728) in the ruminal epithelium, respectively. The ruminal weight was positively correlated to the relative mitochondrial DNA copy number (p < 0.05, r = 0.631) and ATP concentration in ruminal epithelium (p < 0.01, r = 0.957), respectively. The ruminal papillae had a significant positive correlation with ATP concentration in ruminal epithelium (p < 0.01, r = 0.770). These results suggested that growth-retarded yaks had a lower VFA metabolism, ketone bodies synthesis, ion absorption, and ATP synthesis in the ruminal epithelium; it also indicated that the growth retardation of yaks is related to the obstruction of cellular ATP synthesis in rumen epithelial cells. Full article

The Flinders Technology Associates (FTA) card, a cotton-based cellulose membrane impregnated with a chaotropic agent, effectively inactivates infectious microorganisms, lyses cellular material, and fixes nucleic acid. The aim of this study is to assess the stability and detection limit of various RNA viruses, especially the avian influenza virus (AIV), Newcastle disease virus (NDV), and African horse sickness virus (AHSV), on the FTA card, which could significantly impact virus storage and transport practices. To achieve this, each virus dilution was inoculated onto an FTA card and stored at room temperature in plastic bags for durations ranging from 1 week to 6 months. Following storage, the target genome was detected using conventional reverse transcription polymerase chain reaction. The present study demonstrated that the detection limit of AIV ranged from 1.17 to 6.17 EID₅₀ values over durations ranging from 1 week to 5 months, while for NDV, it ranged from 2.83 to 5.83 ELD₅₀ over the same duration. Additionally, the detection limit of AHSV was determined as 4.01 PFU for both 1 and 2 weeks, respectively. Based on the demonstrated effectiveness, stability, and safety implications observed in the study, FTA cards are recommended for virus storage and transport, thus facilitating the molecular detection and identification of RNA viral pathogens. Full article

The production of small ruminant autochthonous breeds in the Centre region of Portugal is practiced in a semi-extensive husbandry system, exposing animals to parasitic infections. The main objective of this study was to estimate the prevalence of lungworm infection and identify risk factors. Fecal samples of 203 goats and 208 sheep from 30 herds were collected per rectum and subjected to the modified Baermann test. The overall prevalence of infection was 57.7%, significantly higher in goats (95.6%) than in sheep (20.7%) (p < 0.001). According to the binary logistic regression model, sheep dewormed with albendazole, mebendazole plus closantel, or ivermectin plus clorsulon presented a risk of Protostrongylidae infection 29.702, 7.426, or 8.720 times higher, respectively, than those dewormed with eprinomectin. Additionally, the presence of gastrointestinal parasites was investigated in 307 fecal samples using Mini-FLOTAC^®. The overall prevalence of infection was 86.3%, also significantly higher in goats (93.2%) than in sheep (79.9%) (p < 0.001). Strongyle-type eggs were the most frequently identified, both in sheep (69.8%) and goats (87.8%), followed by Eimeria oocysts (40.3% in sheep and 68.9% in goats). Considering the high prevalence and the burden of lungworm parasitic infection, it is urgent to determine its economic impact and the repercussions in animal health in the Centre region of Portugal to establish appropriate therapeutic guidelines. Full article

Understanding the spatial variation and formation mechanism of biological diversity is a hot topic in ecological studies. Comparing with α diversity, β diversity is more accurate in reflecting community dynamics. During the past decades, β diversity studies usually focused on plants, mammals, and birds. Studies of amphibian β diversity in montane ecosystems, in particular, tadpoles, are still rare. In this study, Mount Emei, located in southwestern China, was selected as the study area. We explored the tadpole β diversity in 18 streams, based on a two-year survey (2018–2019). Our results indicated a high total β diversity in tadpole assemblages, which was determined by both turnover and nestedness processes, and the dominant component was turnover. Both the total β diversity and turnover component were significantly and positively correlated with geographical, elevational, and environmental distances, but no significant relationship was detected between these and the nestedness component. Moreover, the independent contributions of river width, current velocity, and chlorophyll α were larger than that of geographical and elevational distance. Overall, tadpole β diversity was determined by both spatial and environmental factors, while the contribution of environmental factors was larger. Future studies can focus on functional and phylogenetic structures, to better understand the tadpole assembly process. Full article

Berberine is an alkaloid used to treat diabetes. This experiment aimed to investigate the effects of berberine supplementation in high-carbohydrate diets on the growth performance, glucose metabolism, bile acid synthesis, liver transcriptome, and intestinal flora of Nile tilapia. The six dietary groups were the C group with 29% carbohydrate, the H group with 44% carbohydrate, and the HB1-HB4 groups supplemented with 25, 50, 75, and 100 mg/kg of berberine in group H. The results of the 8-week trial showed that compared to group C, the abundance of Bacteroidetes was increased in group HB2 (p < 0.05). The cholesterol-7α-hydroxylase (CYP7A1) and sterol-27-hydroxylase (CYP27A1) activities were decreased and the expression of FXR was increased in group HB4 (p < 0.05). The pyruvate carboxylase (PC) and phosphoenolpyruvate carboxykinase (PEPCK) activities was decreased in group HB4 (p < 0.05). The liver transcriptome suggests that berberine affects carbohydrate metabolic pathways and primary bile acid synthesis pathways. In summary, berberine affects the glucose metabolism in tilapia by altering the intestinal flora structure, enriching differentially expressed genes (DEGs) in the bile acid pathway to stimulate bile acid production so that it promotes glycolysis and inhibits gluconeogenesis. Therefore, 100 mg/kg of berberine supplementation in high-carbohydrate diets is beneficial to tilapia. Full article

The purpose of this study was to characterize in detail the reproductive strategy, course of embryogenesis, and development of larvae in three species of fishes of the genus Cichlasoma: the green terror (Andinoacara rivulatus), the red discus (Symphysodon discus), and the jaguar cichlid (Parachromis managuensis). Eggs for the study were obtained from five pairs of each species (300 eggs from each female) and incubated at 26 °C. The developing eggs were observed under a microscope (Carl Zeiss Stereo Discovery. V12 and Nikon 2000SE software (NIS-Elements F 4.30.01 64-bit) from fertilization to larval hatching until complete yolk-sac resorption. The largest average number of eggs per female was found in the jaguar cichlid ($\overline{\mathrm{x}}$ = 2991 eggs), a smaller average number of eggs was shown in the green terror ($\overline{\mathrm{x}}$ = 922 eggs), and the red discus showed the smallest average number of eggs ($\overline{\mathrm{x}}$ = 300 eggs). There were significant differences in the sizes of the eggs of the studied species: jaguar cichlid eggs were the smallest (1.060 ± 0.05 mm³), red discus eggs were larger (1.070 ± 0.07 mm³), and green terror eggs were the largest (1.365 ± 0.16 mm³). The embryogenesis time in the red discus was 2132 °H (82 Hpf), in the green terror it was 2158 °H (83 Hpf), and the longest in the jaguar cichlid was 2470 °H (87 Hpf). At the end of embryogenesis, the average size of the larvae after hatching was measured (red discus $\overline{\mathrm{x}}$ = 4.346 mm, green terror $\overline{\mathrm{x}}$ = 5.203 mm, and jaguar cichlid $\overline{\mathrm{x}}$ = 5.301 mm) and the time of yolk-sac resorption from the moment of hatching to the transition from endogenous to exogenous feeding was determined (jaguar cichlid 5 days, green terror 6 days, and red discus 3 days). The results of this study may contribute to the development of reproductive biotechnology for the studied fishes that could be used in aquaculture and, thus, help protect them in their natural habitats. Full article

There are limited studies on the factors affecting the success of ram epididymal spermatozoa (REPS) cryopreservation. On this note, the current study assessed the influence of three commercial soy lecithin-based semen extenders, AndroMed^® (AND), BioXcell^® (BIO), and OviXcell^® (OVI), and two concentrations (400 × 10⁶ vs. 200 × 10⁶ spermatozoa/mL) on the pre-freeze and post-thaw quality of REPS. The REPS were retrieved from nine adult rams’ testes and diluted with each of the three extenders to both concentrations. Straws were frozen manually. Standard motility (SMP) and kinematic parameters (KPs) were assessed via a CASA, while spermatozoa viability, morphology, and acrosomal integrity were assessed via the Kovács–Foote staining technique. The concentration did not significantly affect the pre-freeze and post-thaw SMP and KPs of REPS. BIO and OVI had significantly higher pre-freeze and post-thaw BCFs, post-thaw VAP, and the percentage of all intact heads than AND. In contrast, AND had a significantly lower percentage of REPS with tail defects than BIO and OVI. The 400 × 10⁶ spermatozoa/mL concentration resulted in a significantly higher percentage of all intact heads than the 200 × 10⁶ spermatozoa/mL concentration. Freezing significantly increased tail defects and decreased the percentage of REPS with distal cytoplasmic droplets. The cryopreservation of REPS at the 400 × 10⁶ spermatozoa/mL concentration is recommended. All three extenders must be optimized to preserve the viability, membrane integrity, and better normal morphology of REPS; the reason for increased tail abnormality after the freezing/thawing process needs to be studied. Full article

Spaghetti meat (SM) is a newly identified muscle abnormality that significantly affects modern broiler chickens, consequently exerting a substantial economic impact on the poultry industry worldwide. However, investigations into the meat quality and the underlying causative factors of SM in broilers remain limited. Therefore, this study was undertaken to systematically evaluate meat quality and muscle fiber characteristics of SM-affected meat. To elucidate the disparities between SM-affected and normal (NO) muscles in broiler chickens reared under identical conditions, we selected 18 SM-affected breast tissues and 18 NO breast tissues from 200 broiler chickens raised according to commercial standards under the same conditions for our study. The results showed that compared with the NO group, the muscle surface of the SM group lost integrity, similar to strip and paste. The brightness and yellowness values were significantly higher than those of the NO group. On the contrary, the shear force and protein were significantly lower in the SM group. Microscopic examination revealed that the muscle fibers in the SM group were lysed, necrotic, and separated from each other, with a large number of neutrophils diffusely distributed on the sarcolemma and endometrium. Thirty-five significantly different metabolites were observed in the breast muscles between both groups. Among them, the top differential metabolites—14,15-DiHETrE, isotretinoin, L-malic acid, and acetylcysteine—were mainly enriched in lipid metabolism and inflammatory pathways, including linoleic acid, arachidonic acid, phenylalanine, and histidine metabolism. Overall, these findings not only offer new insights into the meat quality and fiber traits of SM but also contribute to the understanding of potential mechanisms and nutritional regulators for SM myopathy. Full article

This experiment aimed to evaluate the effects of supplementing tributyrin (TB) on the meat quality characteristics of foreshank muscle of weaned lambs. A total of 30 healthy weaned Small-Tailed Han female lambs with body weights ranging from 23.4 to 31.6 kg were selected and randomly divided into five groups, and each group consisted of 6 lambs. The control group was fed a basic total mixed ration, while other groups were fed the same ration supplemented with 0.5, 1.0, 2.0, and 4.0 g/kg TB, respectively. The experiment lasted 75 d, including 15 d of adaptation. Foreshank muscle obtained at the same position from each lamb was used for chemical analysis and sensory evaluation. The results showed that supplementing TB increased the muscle contents of ether extract (p = 0.029), calcium (p = 0.030), phosphorus (p = 0.007), and intermuscular fat length (p = 0.022). Besides, TB increased the muscle pH (p = 0.001) and redness (p < 0.001) but reduced the lightness (p < 0.001), drip loss (p = 0.029), cooking loss (p < 0.001), shear force (p = 0.001), hardness (p < 0.001), cohesiveness (p < 0.001), springiness (p < 0.001), gumminess (p < 0.001), and chewiness (p < 0.001). In addition, TB increased the muscle content of inosine-5′-phosphate (p = 0.004). Most importantly, TB increased the muscle contents of essential amino acids (p < 0.001). Furthermore, TB increased the saturated fatty acids level in the muscle (p < 0.001) while decreasing the unsaturated fatty acids content (p < 0.001). In conclusion, supplementing TB could influence the meat quality of foreshank muscle of weaned lambs by modifying the amino acid and fatty acid levels. Full article

For wild animals, being in captivity in wildlife centers can cause considerable stress. Therefore, it is necessary to establish and validate non-invasive tools to measure chronic stress during rehabilitation. Eight Common Buzzards which lived in permanent husbandry were placed individually into prepared aviaries and their feces were collected before, during and after a stress event for biological validation over a period of seven days. The extracted fecal glucocorticoid metabolites (fGCMs) were analyzed with three different enzyme immune assays (EIA) to find the most suitable one. Additionally, we aimed to investigate the stability of fGCM levels after defecation because further metabolization by bacterial enzymes can lead to changed results. The Cortisone-EIA performed best in males and females and showed that the stress event led to an fGCM increase of 629% (557% in females and 702% in males) in relation to basal values. We found no significant differences between the sexes, but observed significant differences between different times of day. FGCM concentration significantly changed after eight hours at room temperature. Our study successfully validated the non-invasive measurement of fGCM as a stress indicator in Common Buzzards and could therefore lay the foundation for future studies providing new insights for animal welfare research in Buzzards. Full article

Mulberry leaves, a common traditional Chinese medicine, represent a potential nutritional strategy to improve the fat profile, also known as the lipo-nutrition, of pork. However, the effects of mulberry leaves on pork lipo-nutrition and the microorganisms and metabolites in the porcine gut remain unclear. In this study, multi-omics analysis was employed in a Yuxi black pig animal model to explore the possible regulatory mechanism of mulberry leaves on pork quality. Sixty Yuxi black pigs were divided into two groups: the control group (n = 15) was fed a standard diet, and the experimental group (n = 45) was fed a diet supplemented with 8% mulberry leaves. Experiments were performed in three replicates (n = 15 per replicate); the two diets were ensured to be nutritionally balanced, and the feeding period was 120 days. The results showed that pigs receiving the diet supplemented with mulberry leaves had significantly reduced backfat thickness (p < 0.05) and increased intramuscular fat (IMF) content (p < 0.05) compared with pigs receiving the standard diet. Lipidomics analysis showed that mulberry leaves improved the lipid profile composition and increased the proportion of triglycerides (TGs). Interestingly, the IMF content was positively correlated with acyl C18:2 and negatively correlated with C18:1 of differential TGs. In addition, the cecal microbiological analysis showed that mulberry leaves could increase the abundance of bacteria such as UCG-005, Muribaculaceae_norank, Prevotellaceae_NK3B31_group, and Limosilactobacillus. Simultaneously, the relative levels of L-tyrosine-ethyl ester, oleic acid methyl ester, 21-deoxycortisol, N-acetyldihydrosphingosine, and mulberrin were increased. Furthermore, we found that mulberry leaf supplementation significantly increased the mRNA expression of lipoprotein lipase, fatty acid-binding protein 4, and peroxisome proliferators-activated receptor γ in muscle (p < 0.01). Mulberry leaf supplementation significantly increased the mRNA expression of diacylglycerol acyltransferase 1 (p < 0.05) while significantly decreasing the expression of acetyl CoA carboxylase in backfat (p < 0.05). Furthermore, mulberry leaf supplementation significantly upregulated the mRNA expression of hormone-sensitive triglyceride lipase and peroxisome proliferator-activated receptor α (p < 0.05) in backfat. In addition, mulberry leaf supplementation led to increased serum leptin and adiponectin (p < 0.01). Collectively, this omic profile is consistent with an increased ratio of IMF to backfat in the pig model. Full article

Mineral metabolism, in particular Ca, and to a lesser extent phosphorus (P) and magnesium (Mg), is altered with the onset of lactation because of extensive irreversible loss to synthesize colostrum and milk. The transient reduction in the concentration of Ca in blood, particularly when it lasts days, increases the risk of mineral-related disorders such as hypocalcemia and, to a lesser extent, hypophosphatemia. Although the incidence of clinical hypocalcemia can be reduced by prepartum dietary interventions, subclinical hypocalcemia remains prevalent, affecting up to 60% of the dairy cows in the first 3 d postpartum. More importantly, strong associations exist between hypocalcemia and increased susceptibility to other peripartum diseases and impaired reproductive performance. Mechanistic experiments have demonstrated the role of Ca on innate immune response in dairy cows, which presumably predisposes them to other diseases. Hypocalcemia is not related to inadequate Ca intake as prepartum diets marginal to deficient in Ca reduce the risk of the disease. Therefore, the understanding of how Ca homeostasis is regulated, in particular how calciotropic hormones such as parathyroid hormone and 1,25-dihydroxyvitamin D₃, affect blood Ca concentrations, gastrointestinal Ca absorption, bone remodeling, and renal excretion of Ca become critical to develop novel strategies to prevent mineral imbalances either by nutritional or pharmacological interventions. A common method to reduce the risk of hypocalcemia is the manipulation of the prepartum dietary cation-anion difference. Feeding acidogenic diets not only improves Ca homeostasis and reduces hypocalcemia, but also reduces the risk of uterine diseases and improves productive performance. Feeding diets that induce a negative Ca balance in the last weeks of gestation also reduce the risk of clinical hypocalcemia, and recent work shows that the incorporation of mineral sequestering agents, presumably by reducing the absorption of P and Ca prepartum, increases blood Ca at calving, although benefits to production and health remain to be shown. Alternative strategies to minimize subclinical hypocalcemia with the use of vitamin D metabolites either fed prepartum or as a pharmacological agent administered immediately after calving have shown promising results in reducing hypocalcemia and altering immune cell function, which might prove efficacious to prevent diseases in early lactation. This review summarizes the current understanding of Ca homeostasis around parturition, the limited knowledge of the exact mechanisms for gastrointestinal Ca absorption in bovine, the implications of hypocalcemia on the health of dairy cows, and discusses the methods to minimize the risk of hypocalcemia and their impacts on productive performance and health in dairy cows. Full article

The present meta-analysis aimed to determine the underlying effects of different saponins extracted from different sources on the production performance, milk yield, digestibility, rumen fermentation, blood metabolites, and nitrogen utilization of ruminants. A total of 26 papers comprising 66 in vivo studies (148 data points of dietary treatments) were evaluated in the present study. The databases were statistically analyzed using the mixed model procedure of SAS, where experiments considered random effects and tannin-related factors were treated as fixed effects. Statistical procedures were then continued in comparing different sources of saponin extract through Mixed Model analysis, where experiments were also random factors and sources of saponin extract were fixed factors. The evidence revealed in the present meta-analysis that saponin supplementation of up to 40 g/kg DM appears to have no detrimental impact on feed intake across ruminant types, suggesting that it does not significantly affect diet palatability. However, the results indicated that there are species-specific responses to saponin supplementation, particularly in relation to palatability and nutrient absorption efficiency, with larger ruminants being better able to tolerate the bitterness induced by saponin extracts. Furthermore, the study found that saponin extracts can influence nutrient digestibility and rumen fermentation dynamics, with different effects observed in large and small ruminants. While some saponin extracts can enhance average daily weight gain and milk yield, others can have adverse effects, highlighting the importance of considering both saponin sources and animal physiological condition when developing nutritional strategies. Additionally, optimization of ruminant production by utilizing saponin extracts is necessary to avoid negative health implications, such as increased blood creatinine levels. Different saponin extracts utilization in ruminant nutrition and environmental management, have a distinct understanding associated to their various bioactive properties. However, among the saponin sources, saponin extracted from Quilaja saponaria is more likely to improve large ruminant production performance while maintaining ruminant health and metabolism, but negatively affect small ruminants. Further research is needed to unravel the intricate effects of different saponin sources on ruminant health and productivity, emphasizing the importance of tailored dietary strategies that consider the unique physiological and metabolic characteristics of the target livestock. Full article

The study was conducted within a well-managed beef cattle operation in northeastern Mexico. Each nest was weighed and dissected to obtain the plant and animal material used to build the nests. The number of materials present per nest and relative frequency were determined. Twenty-one building materials were used. Over the years, Muhlenbergia torreyi represented 85.5% of the total biomass of the nests, and Aristida longiseta, Bouteloua gracilis, Brickellia canescens, Purshia mexicana and Cirsium ehrenbergii constituted 2.45, 2.80, 2.44, 1.34 and 1.11% of the total biomass, respectively. The above-mentioned grasses represented 95.62% of the total biomass. Material of animal origin was horse and cow hair, which represented 0.84 and 0.58% of the total biomass, respectively. It was concluded that, at the study site, Muhlenbergia torreyi was a key nest-building material for the Worthen sparrow nest. Full article

Mammary cancer is a frequent disease in female dogs, where a high proportion of cases correspond to malignant tumors that may exhibit drug resistance. Within the mammary tumor microenvironment, there is a cell subpopulation called cancer stem cells (CSCs), which are capable of forming spheres in vitro and resisting anti-tumor treatments, partly explaining the recurrence of some tumors. Previously, it has been described that spheres derived from canine mammary carcinoma cells CF41.Mg and REM 134 exhibit stemness characteristics. Melatonin has shown anti-tumor effects on mammary tumor cells; however, its effects have been poorly evaluated in canine mammary CSCs. This study aimed to analyze the effect of melatonin on the chemoresistance exhibited by stem-like neoplastic cells derived from canine mammary carcinoma to cytotoxic drugs such as doxorubicin and mitoxantrone. CF41.Mg and REM 134 cells were cultured in high-glucose DMEM supplemented with fetal bovine serum and L-glutamine. The spheres were cultured in ultra-low attachment plates in DMEM/F12 medium without fetal bovine serum and with different growth factors. The CD44⁺/CD24^−/low phenotype was analyzed by flow cytometry. The viability of sphere-derived cells (MTS reduction) was studied in the presence of melatonin (0.1 or 1 mM), doxorubicin, mitoxantrone, and luzindole. In addition, the gene (RT-qPCR) of the multidrug resistance bombs MDR1 and ABCG2 were analyzed in the presence of melatonin. Both cell types expressed the MT1 gene, which encodes the melatonin receptor MT1. Melatonin 1 mM does not modify the CD44⁺/CD24^−/low phenotype; however, the hormone reduced viability (p < 0.0001) only in CF41.Mg spheres, without inducing an additive effect when co-incubated with cytotoxic drugs. These effects were independent of the binding of the hormone to its receptor MT1, since, by pharmacologically inhibiting them, the effect of melatonin was not blocked. In CF41.Mg spheres, the relative gene expression of ABCG2 and MDR1 was decreased in response to the hormone (p < 0.001). These results indicate that melatonin negatively modulates the cell survival of spheres derived from CF41.Mg cells, in a way that is independent of its MT1 receptor. These effects did not counteract the resistance to doxorubicin and mitoxantrone, even though the hormone negatively regulates the gene expression of MDR1 and ABCG2. Full article

The gastrointestinal tract plays crucial roles in the digestion and absorption of nutrients, as well as in maintenance of a functional barrier. The development and maturation of the intestine is important for piglets to maintain optimal growth and health. Polyamines are necessary for the proliferation and growth of enterocytes, which play a key role in differentiation, migration, remodeling and integrity of the intestinal mucosa after injury. This review elaborates the development of the structure and function of the intestine of piglets during embryonic, suckling and weaning periods, the utilization and metabolism of polyamines in the intestine, as well as the role of polyamines in intestinal development and mucosal repair. The nutritional intervention to improve intestinal development and functions by modulating polyamine metabolism in piglets is also put forward. These results may help to promote the adaption to weaning in pigs and provide useful information for the development and health of piglets. Full article

A pig inventory is a crucial component of achieving precise and large-scale farming. In complex pigsty environments, due to pigs’ stress reactions and frequent obstructions, it is challenging to count them accurately and automatically. This difficulty contrasts with most current deep learning studies, which rely on overhead views or static images for counting. This research proposes a video-based dynamic counting method, combining YOLOv7 with DeepSORT. By utilizing the YOLOv7 network structure and optimizing the second and third 3 × 3 convolution operations in the head network ELAN-W with PConv, the model reduces the computational demand and improves the inference speed without sacrificing accuracy. To ensure that the network acquires accurate position perception information at oblique angles and extracts rich semantic information, we introduce the coordinate attention (CA) mechanism before the three re-referentialization paths (REPConv) in the head network, enhancing robustness in complex scenarios. Experimental results show that, compared to the original model, the improved model increases the mAP by 3.24, 0.05, and 1.00 percentage points for oblique, overhead, and all pig counting datasets, respectively, while reducing the computational cost by 3.6 GFLOPS. The enhanced YOLOv7 outperforms YOLOv5, YOLOv4, YOLOv3, Faster RCNN, and SSD in target detection with mAP improvements of 2.07, 5.20, 2.16, 7.05, and 19.73 percentage points, respectively. In dynamic counting experiments, the improved YOLOv7 combined with DeepSORT was tested on videos with total pig counts of 144, 201, 285, and 295, yielding errors of -3, -3, -4, and -26, respectively, with an average accuracy of 96.58% and an FPS of 22. This demonstrates the model’s capability of performing the real-time counting of pigs in various scenes, providing valuable data and references for automated pig counting research. Full article

In response to the high breakage rate of pigeon eggs and the significant labor costs associated with egg-producing pigeon farming, this study proposes an improved YOLOv8-PG (real versus fake pigeon egg detection) model based on YOLOv8n. Specifically, the Bottleneck in the C2f module of the YOLOv8n backbone network and neck network are replaced with Fasternet-EMA Block and Fasternet Block, respectively. The Fasternet Block is designed based on PConv (Partial Convolution) to reduce model parameter count and computational load efficiently. Furthermore, the incorporation of the EMA (Efficient Multi-scale Attention) mechanism helps mitigate interference from complex environments on pigeon-egg feature-extraction capabilities. Additionally, Dysample, an ultra-lightweight and effective upsampler, is introduced into the neck network to further enhance performance with lower computational overhead. Finally, the EXPMA (exponential moving average) concept is employed to optimize the SlideLoss and propose the EMASlideLoss classification loss function, addressing the issue of imbalanced data samples and enhancing the model’s robustness. The experimental results showed that the F1-score, mAP50-95, and mAP75 of YOLOv8-PG increased by 0.76%, 1.56%, and 4.45%, respectively, compared with the baseline YOLOv8n model. Moreover, the model’s parameter count and computational load are reduced by 24.69% and 22.89%, respectively. Compared to detection models such as Faster R-CNN, YOLOv5s, YOLOv7, and YOLOv8s, YOLOv8-PG exhibits superior performance. Additionally, the reduction in parameter count and computational load contributes to lowering the model deployment costs and facilitates its implementation on mobile robotic platforms. Full article

Few studies analyze the morphology and anatomy of the bat skull, and most of them are incomplete. Some of the difficulties stem from the fact that, in the representatives of the order Chiroptera, the interosseous sutures disappear by fusing together before active flight begins, which takes place over only a few months. This study presents a detailed morphological and anatomical description of the skull of a juvenile specimen of Myotis myotis (Borkhausen, 1797). Juvenile skulls are difficult to preserve and often incomplete. Previously inconsistent terminology related to bones, sutures, and other cranial structures was unified, which will provide insight on the distribution of each structure in both juvenile and adult specimens to be investigated. The description fill in the gaps in knowledge about the cranial structures of Myotis myotis and the representatives of the family Vespertilionidae. This will allow for precise descriptions of the skulls of bats. Full article

Tumor cells can induce important cellular and molecular modifications in the tissue or host where they grow. The idea that the host and tumor interact with each other has led to the concept of a tumor microenvironment, composed of immune cells, stromal cells, blood vessels, and extracellular matrix, representing a unique environment participating and, in some cases, promoting cancer progression. The study of the tumor immune microenvironment, particularly focusing on the role of tumor-infiltrating lymphocytes (TILs), is highly relevant in oncology due to the prognostic and therapeutic significance of TILs in various tumors and their identification as targets for therapeutic intervention. Canine splenic hemangiosarcoma (HSA) is a common tumor; however, its immune microenvironment remains poorly understood. This retrospective study aimed to characterize the histological and immunohistochemical features of 56 cases of canine splenic HSA, focusing particularly on tumor-infiltrating lymphocytes (TILs). We assessed the correlations between the lymphocytic response, the macroscopic and histological characteristics of the tumor, and the survival data. Our study demonstrated that FoxP3 distribution was associated with tumor-related death and survival, while the CD20 count was associated with metastasis. This study provides an in-depth characterization of the tumor immune microenvironment in canine splenic HSA and describes potential prognostic factors. Full article