Animals, Volume 14, Issue 5 (March-1 2024) – 163 articles

Cultured meat production relies on various cell types, including muscle stem cells (MuSCs), embryonic stem cell lines, induced pluripotent cell lines, and naturally immortalized cell lines. MuSCs possess superior muscle differentiation capabilities compared to the other three cell lines, making them key for cultured meat development. Therefore, to produce cultured meat using MuSCs, they must first be effectively separated from muscles. At present, the methods used to isolate MuSCs from muscles include (1) the pre-plating method, using the ability of cells to adhere differently, which is a biological characteristic of MuSCs; (2) the density gradient centrifugation method, using the intrinsic density difference of cells, which is a physical characteristic of MuSCs; and (3) fluorescence- and magnetic-activated cell sorting methods, using the surface marker protein on the cell surface of MuSCs, which is a molecular characteristic of MuSCs. Further efficient and valuable methods for separating MuSCs are expected to be required as the cell-based cultured meat industry develops. Thus, we take a closer look at the four methods currently in use and discuss future development directions in this review. Full article

This study aimed to investigate the interactions between corn particle size (PS) and conditioning temperature (CT) on the performance, carcass traits, intestinal morphology, and immune responses in broilers fed a corn-soybean meal-based diet. A total of 360 one-day-old male broiler chicks (Ross 308) were randomly allocated into six dietary treatments in a 2 × 3 factorial arrangement, consisting of two corn PS (finely ground with geometric mean diameter (GMD) of 357 µm (PS_F) vs. coarsely ground corn with GMD of 737 µm (PS_C), and three CT [unconditioned (CT_U), conditioned at 75 °C (CT₇₅) and 90 °C (CT₉₀)]. Birds were accommodated in 30 pens with five replicates and 12 chicks per each pen. There was no interaction between corn PS and CT on the growth performance and immune response of broilers at any growth phases. However, during the starter (0–10 days) period, the average daily weight gain (ADWG) and feed conversion ratio (FCR) of PS_F-fed birds were significantly improved compared to those fed PS_C (p < 0.05). During the starter (0–10 days) and grower (11–24 days) periods, increasing the conditioning temperature of corn increased the ADWG, while in the starter phase only the CT₇₅ caused a lower FCR (p < 0.05). Broilers fed PS_F corn showed the lowest FCR during the finisher (25–42 days) period compared to those fed PS_C (p < 0.05). Conditioning corn at 75 °C reduced FCR during the finisher (25–42 days) period compared to the birds fed CT_U and CT₉₀ corn (p < 0.05). In whole experimental periods (1–42 days), PS_F and CT₇₅ treatment increased the ADWG compared to the PS_C and CT_U (p < 0.05). The CT₇₅ treatment improved primary total anti-sheep red blood cell (SRBCs) titer (IgT) and IgM and secondary IgT and IgG responses compared to the other experimental groups (CT_U and CT₉₀) (p < 0.05). No significant PS × CT interaction was found on the Newcastle disease (ND) antibody titer of broiler chickens (p > 0.05). Feeding CT₇₅ corn reduced duodenum and jejunum relative lengths compared to the birds fed diets containing CT_U corn. Significant PS × CT interactions (p < 0.05) were observed for villus height, villus height to crypt depth, crypt depth, muscle thickness, and absorption surface area of the jejunum. The highest carcass yield was observed in the PS_F-CT₇₅ group (p < 0.05). In conclusion, the use of finely ground corn (PSF) conditioned at 75 °C (CT75) was beneficial to growth performance, development of the digestive tract, jejunum histomorphometry and the immune responses of broilers. Full article

Working equids are often used to exhaustion, living and dying in conditions below minimal welfare standards. Due to their poor welfare status, euthanasia should be considered in certain conditions. The study aimed to describe the population and the disease frequency of the working equids admitted at an equine clinic in Cairo (i.e., Egypt Equine Aid (EEA)) from 2019 to 2022 and identify possible associations between physical parameters at admission and the outcome. Records of 1360 equids admitted at EEA were reviewed. The majority of the admitted equids were horses (65.6%), followed by donkeys (33%), in particular stallions (68.7%), from 1 to 15 years old (74.8%). Hospitalisation was mainly due to wounds (28.9%), orthopaedic problems (27.4%), colic (8.5%), or infectious diseases (7.4%). The majority of the equids were discharged, but 5.1% died on their own, without human intervention, and 23% were euthanised. Text mining revealed the anamnesis’s most frequent words were ‘accident’, ‘lameness’, and ‘wound’. In addition, owners sometimes reported using inappropriate remedies (e.g., firing) before hospitalisation. Multivariable ordinal regression analysis performed between physical parameters and the outcome (ordered based on severity: discharged, euthanasia, and dead) revealed that sex (male vs. female: OR = 1.33; p < 0.05), colour of the mucous membrane (pathological vs. physiological: OR = 1.72; p < 0.01), and capillary refill time (pathological vs. physiological: OR = 1.42; p = 0.02) increased the likelihood of a non-survival outcome. In conclusion, early euthanasia should be considered for these equids, to minimise prolonged suffering. Moreover, owners’ education is recommended to guarantee minimal welfare standards to the working equids. Full article

This review paper provides an in-depth analysis of three critical metabolic diseases affecting dairy cattle such as subacute ruminal acidosis (SARA), ketosis, and hypocalcemia. SARA represents a disorder of ruminal fermentation that is characterized by extended periods of depressed ruminal pH below 5.5–5.6. In the long term, dairy herds experiencing SARA usually exhibit secondary signs of the disease, such as episodes of laminitis, weight loss and poor body condition despite adequate energy intake, and unexplained abscesses usually 3–6 months after an episode of SARA. Depressed milk-fat content is commonly used as a diagnostic tool for SARA. A normal milk-fat test in Holstein dairy cows is >4%, so a milk-fat test of <3% can indicate SARA. However, bulk tank testing of milk fat is inappropriate to diagnose SARA at the herd level, so when >4 cows out of 12 and <60 days in milk are suspected to have SARA it can be considered that the herd has a problem. The rapid or abrupt introduction of fresh cows to high-concentrate diets is the most common cause of SARA. Changes in ruminal bacterial populations when exposed to higher concentrate rations require at least about 3 weeks, and it is recommended that concentrate levels increase by no more than 400 g/day during this period to avoid SARA. Ketosis, a prevalent metabolic disorder in dairy cattle, is scrutinized with a focus on its etiological factors and the physiological changes leading to elevated ketone bodies. In total mix ration-fed herds, an increased risk of mastitis and reduced fertility are usually the first clinical signs of ketosis. All dairy cows in early lactation are at risk of ketosis, with most cases occurring in the first 2–4 weeks of lactation. Cows with a body condition score ≥3.75 on a 5-point scale at calving are at a greater risk of ketosis than those with lower body condition scores. The determination of serum or whole blood acetone, acetoacetate, beta-hydroxybutyrate (BHB) concentration, non-esterified fatty acids (NEFA), and liver biopsies is considered the best way to detect and monitor subclinical ketosis, while urine or milk cowside tests can also be used in on-farm monitoring programs. Concentrations >1.0 mmol/L or 1.4 mmol/L blood or serum BHB are considered diagnostic of subclinical ketosis. The standard threshold used for blood is 1.2 mmol/L, which corresponds to thresholds of 100 mcmol/L for milk and 15 mg/dL for urine. Oral administration of propylene glycol (250–400 g, every 24 h for 3–5 days) is the standard and most efficacious treatment, as well as additional therapy with bolus glucose treatment. Hypocalcemia is a disease of adult dairy cows in which acute hypocalcemia causes acute to peracute, afebrile, flaccid paralysis that occurs most commonly at or soon after parturition. Dairy cows are at considerable risk for hypocalcemia at the onset of lactation, when daily calcium excretion suddenly increases from about 10 g to 30 g per day. Cows with hypocalcemia have a more profound decrease in blood calcium concentration—typically below 5.5 mg/dL. The prevention of parturient paresis has been historically approached by feeding cows low-calcium diets during the dry period. Negative calcium balance triggers calcium mobilization before calving and better equips the cow to respond to the massive calcium needs at the onset of lactation. Calcium intake must be limited to <20 g per day for calcium restriction to be effective. The most practical and proven method for monitoring hypocalcemia is by feeding cows an acidogenic diet for ~3 weeks before calving. Throughout the review, emphasis is placed on the importance of early diagnosis and proactive management strategies to mitigate the impact of these metabolic diseases on dairy cattle health and productivity. The comprehensive nature of this paper aims to serve as a valuable resource for veterinarians, researchers, and dairy farmers seeking a deeper understanding of these prevalent metabolic disorders in dairy cattle. Full article

Insectivorous bats are generalist predators and can flexibly respond to fluctuations in the distribution and abundance of insect prey. To better understand the effects of bats on arthropod pests, the types of pests eaten by bats and the response of bats to insect prey need to be determined. In this study, we performed DNA metabarcoding to examine prey composition and pest diversity in the diets of four insectivorous species of bats (Hipposideros armiger, Taphozous melanopogon, Aselliscus stoliczkanus, and Miniopterus fuliginosus). We evaluated the correlation between bat activity and insect resources and assessed dietary niche similarity and niche breadth among species and factors that influence prey consumption in bats. We found that the diets of these bats included arthropods from 23 orders and 200 families, dominated by Lepidoptera, Coleoptera, and Diptera. The proportion of agricultural pests in the diet of each of the four species of bats exceeded 40% and comprised 713 agricultural pests, including those that caused severe economic losses. Bats responded to the availability of insects. For example, a higher abundance of insects, especially Lepidoptera, and a higher insect diversity led to an increase in the duration of bat activity. In areas with more abundant insects, the number of bat passes also increased. The dietary composition, diversity, and niches differed among species and were particularly significant between H. armiger and T. melanopogon; the dietary niche width was the greatest in A. stoliczkanus and the narrowest in H. armiger. The diet of bats was correlated with their morphological and echolocation traits. Larger bats preyed more on insects in the order Coleoptera, whereas the proportion of bats consuming insects in the order Lepidoptera increased as the body size decreased. Bats that emitted echolocation calls with a high peak frequency and duration preyed more on insects in the order Mantodea. Our results suggest that dietary niche differentiation promotes the coexistence of different bat species and increases the ability of bats to consume insect prey and agricultural pests. Our findings provide greater insights into the role of bats that prey on agricultural pests and highlight the importance of combining bat conservation with integrated pest management. Full article

Pampus is a widespread species of fish in the western Pacific and Indian Oceans that has significant commercial worth. Its evolutionary history and phylogenetics are still poorly understood, and details on its intraspecific taxonomy are debatable, despite some morphological and molecular research. Here, we analyzed this species using skeletal structure data as well as nuclear (S7 gene) and mitochondrial genetic information (COI, D-loop and mitogenomes). We found that the genetic distance between P. argenteus and P. echinogaster was much smaller than that between other Pampus species, and both maximum likelihood and Bayesian phylogenetic trees yielded almost identical tree topologies. An additional and adjacent M repeat was found in the downstream region of the IQM gene cluster of P. argenteus and P. echinogaster, and the trnL2 gene of P. minor was translocated. The genus Pampus experienced early rapid radiation during the Palaeocene with major lineages diversifying within a relatively narrow timescale. Additionally, three different methods were conducted to distinguish the genus Pampus species, proving that P. argenteus and P. echinogaster are the same species, and P. liuorum is speculated to be a valid species. Overall, our study provides new insights not only into the evolutionary history of Pampus but its intraspecific taxonomy as well. Full article

We have conducted a 10-year-long coprological study of the animals housed in two zoological institutions (ZooAquarium and Faunia, Madrid, Spain) to assess the parasite biodiversity, prevalence, and their relation with host class, diet, and enclosure type (soil type and level of isolation from wild fauna). A total of 4476 faecal samples from 132 mammal species and 951 samples from 86 avian species were examined. The results indicated that only 12.8% of avian species had parasites at least once during the study period, whereas 62.1% of mammal species tested positive. Predominantly, protists (Entamoeba, flagellates, and ciliates) and nematodes (mainly Trichuris) were identified in the findings. Carnivorous species were primarily infected by nematodes, while herbivorous and omnivorous species were mainly infected by protists. The number of infected herbivorous and omnivorous species was significantly greater than carnivorous species. Differences were observed based on soil type (artificial, natural, mixed) and isolation level (isolated/accessible), but these differences were not statistically significant. Several parasites (Entamoeba spp., Giardia spp., Balantidoides coli, Trichuris spp.) could potentially be transmitted between humans and some mammals and birds. Regular animal analyses and a personnel health program in the institutions would minimise transmission risks between zoo animals, wildlife, and humans. Full article

Metacarpophalangeal joint region pain is a common cause of lameness in racehorses. Radiological abnormalities in the sagittal ridge (SR) of the third metacarpal bone have been associated with joint effusion, lameness and reduced sales prices. The aims were to describe computed tomographic (CT) appearance of the SR in racehorses, and to document the progression of these findings over three assessments. Forty yearlings were enrolled at the first examination (time 0). Re-examinations were performed twice, approximately six months apart on 31 (time 1) and 23 (time 2) horses, respectively. Computed tomographic examinations of both metacarpophalangeal regions were performed with the horses in a standing position. Computed tomographic reconstructions were analysed subjectively and objectively. The mean Hounsfield Unit values (Hus) of eight radial segments and location, size and shape of hypoattenuating lesions were recorded. Mean Hus at time 1 were higher than at time 0. There was no difference between mean HU at times 1 and 2. The mean HU values of the dorsal half were higher in the right forelimbs and in fillies. Hypoattenuation was identified in 33/80 (41.3%) limbs at time 0, in 22/62 (35.5%) limbs at time 1 and in 14/46 (30.4%) limbs at time 2. All hypoattenuations were located in the dorsodistal aspect of the SR. The most common shapes were hypoattenuating lesions elongated proximodistally and those extending towards trabecular bone. An increase in attenuation of the SR occurred in the first six months of training. Hypoattenuating lesions could decrease in size and could resolve during early training. In this population, these lesions were not associated with lameness. Full article

Oxidative stress resulting from an imbalance between oxidants and antioxidants can cause damage to certain cellular components. Purple Napier grass, a semi-dwarf variety, is characterized by its purple leaves and contains anthocyanins, which provide it with antioxidant properties. This study examined the effects of feeding purple Napier grass (“Prince”) silage to lactating dairy goats on blood antioxidant activity, milk yield, and milk quality. Eighteen female Saanen crossbred goats, weighing 52.34 ± 2.86 kg and producing milk for 14 ± 2 days, were systematically divided into three groups based on their lactation period in the previous cycle as follows: early, mid, and late lactation. In a randomized complete block design (RCBD), treatments were randomly allocated to six animals in each block. The dairy goats were fed a total mixed ration (TMR) consisting of the three following treatments: control (100% Napier Pakchong 1 grass silage), 50% (a 50% replacement of the control with purple Napier grass silage), and 100% (100% purple Napier grass silage). The results show that goats who were fed a diet including 100% purple Napier grass silage showed higher levels of certain milk contents, especially with regard to lactose, when compared to those who were fed a control diet, as well as a diet with a 50% replacement of purple Napier grass silage. The somatic cell count (SCC) of these goats was reduced. In terms of antioxidant activity, dairy goats who were fed 100% purple Napier grass silage showed higher levels of enzymes in both plasma and milk, including glutathione s-transferase, total antioxidant capacity, superoxide dismutase, and 2,2-diphenyl-1-picrylhydrazyl radical, compared to the control group and the 50% replacement group. The plasma and milk of these goats showed lower levels of malondialdehyde. The dairy goats who were fed a 100% purple Napier grass silage diet showed higher concentrations of anthocyanins, including C3G, P3G, Peo3G, M3G, Cya, Pel, and total anthocyanins in milk, when compared to the control group and the 50% replacement group. The increased replacement of purple Napier grass silage led to significant differences in lactose levels, somatic cell count, glutathione S-transferase, total antioxidant capacity, superoxide dismutase, 2,2-diphenyl-1-picrylhydrazyl radical, and the composition of anthocyanins. This study provides evidence to support the use of purple Napier grass silage as a beneficial source of roughage for lactating dairy goats. Full article

This study aimed to investigate the appearance and frequencies of the Booroola polymorphism of the bone morphogenetic protein receptor 1b (BMPR1B) gene (FecB) and the Embrapa polymorphism of the growth differentiation factor 9 (GDF9) gene (FecG^E) in sheep in Thailand. A total of 454 crossbred sheep blood samples were collected from four provinces in Thailand during August 2022 to July 2023. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was used to identify the FecB and FecG^E genotypes. The history of ewe birth types was collected from the owners to analyze the association between fecundity (Fec) genotypes and the history of birth types. The genotypic frequencies of FecB for homozygous genotype (B/B), heterozygous genotype (+/B), and wildtype (+/+) were 0.22%, 1.54%, and 98.24%, respectively. Meanwhile, the genotypic frequencies of FecG^E for homozygous genotype (E/E), heterozygous genotype (+/E), and wildtype (+/+) were 0.00%, 2.42%, and 97.58%, respectively. Furthermore, three ewes exhibited both FecB and FecG^E genotypes. Fisher’s exact test revealed that possession of the FecB genotype was associated with multiple births (p < 0.01). Both FecB and FecG^E mutations were identified in crossbred sheep in Thailand. Sheep containing FecB allele could be alternative candidates to be selected to improve the prolificacy of crossbred sheep in Thailand. Full article

With the results of a survey presented in this paper, we provide insight into public attitudes towards dealing with wildlife. Based on 1569 data sets derived from participating stakeholders, we inquired about the individual experience the participants had made with wild animals, and asked about their personal engagement, attitude towards management, and emotions involved and tried to evaluate basic contextual knowledge. As a result, we discovered a positive effect showing that a strong opinion about dealing with wildlife is associated with increasing contextual knowledge. People that are experienced in and engaged in wildlife conservation expressed significantly stronger positive emotions in this context. We conclude that education is essential in dealing with wildlife responsibly and that positive emotions are a main trigger for such engagement. The results of the survey underline that a combination of contextual knowledge and a positive attitude towards wildlife leads to a higher awareness of possible conflicts between humans and wildlife. Furthermore, these criteria are crucial when developing strategies that strive for a sustainable coexistence. Full article

Reproductive biotechnologies can be used as a supporting tool, through gamete conservation and in vitro embryo production, in the preservation of invaluable and irreplaceable animal genetic resources. In the present study, immature mouflon cumulus–oocyte complexes (COCs) collected from ovariectomized female ovaries underwent short- or long-term conservation (24 h maintained in Earle’s/Hank’s (EH) medium or vitrification) under field conditions and afterwards transported to the laboratory where they were cultured for in vitro maturation (IVM) and assessed for oocyte meiotic competence and bioenergetic–oxidative status. Utilization of both storage techniques led to COC morphology preservation, as well as cumulus expansion and oocyte meiotic resumption after the IVM procedure. Quantitative bioenergetic–oxidative parameters were reduced in vitrified oocytes compared with EH ones. Immature COC storage needs to be optimized in both domesticated and non-domesticated sheep as a part of the strategy to avoid the loss of valuable genotypes of these animal species. Full article

N-alkanes offer a promising approach for assessing the nutritional contribution of external sources to the diets of laying hens in free-range production systems. However, traditional laboratory methods, involving extraction, purification and gas chromatographic analysis, are both economically burdensome and time-consuming. Near-infrared spectroscopy (NIRS) is emerging as a viable alternative, with varying degrees of accuracy depending on the chemical nature and concentration of the component of interest. In our research, we focus on the accuracy of NIRS in predicting the concentrations of n-alkanes (C25–C33) in excreta under simulated free-range conditions with two different diets: one containing a commercial feed with minimal n-alkane content and another containing 1% alfalfa on top of the commercial feed. Spectra processing and calibration were tailored for each n-alkane, with NIRS performance influenced by diet type. Notably, plant predictions using NIR-generated data were consistent with laboratory results, despite a slight tendency toward overestimation (3.40% using the NIRS-generated C25-C29-C33 combination versus 2.80% using laboratory analysis). This indicates the potential of NIRS as an efficient tool to assess n-alkanes in excreta of laying hens and, consequently, the nutritional contribution of the free-range environment, providing rapid and cost-effective results. Full article

Miranda’s donkey is an endangered, autochthone breed from Northern Portugal. Understanding the physiological and pathological conditions of Miranda’s donkey is crucial for the conservation of the breed. Our study aimed to establish reference intervals (RIs) for blood biochemistry parameters and to evaluate the influence of gender and age on these parameters. Blood samples from 75 clinically healthy animals were analyzed for 21 blood biochemistry parameters using Respons^® 920 and Start^® 4-Diagnostica-Stago. RIs were calculated according to the ASVCP guidelines, utilizing Reference Advisor software V. 2.1 and the statistical program SPSS version 29 to analyze the effects of gender and age. Significant gender-related differences (p < 0.05) were observed in cholesterol, chlorine, creatinine (CREA), glucose (GLU), sodium, and triglycerides (with higher values in females) and in aspartate aminotransferase, creatine phosphokinase (CK), gamma-glutamyl transferase, potassium, magnesium (Mg), and urea (with higher values in males). Age-related differences (p < 0.05) were noted for alkaline phosphatase, CK, fibrinogen, GLU, and phosphorus (higher in young animals) and for globulin, Mg, and total protein (higher in adults). The RIs described here are invaluable for assessing and monitoring the health status of individual animals and herds. Our study highlights the importance of considering gender and age in the interpretation of biochemical parameters, providing critical insights for the conservation and management of this endangered breed. Full article

Jumps racing is a form of Thoroughbred horse racing that involves hurdles and steeples and typically longer distances, and heavier weights compared with flat racing, which does not incorporate obstacles. In Australia, jumps racing is carried out only in Victoria, one of eight states and territories. The continuation of jumps racing is contentious due to the higher risk of fatalities, falls and injuries for horses, compared with flat racing. While measures have been introduced by the industry to improve the safety of riders and horses, the rates of fatalities, falls and injuries in horses participating in jumps races have not been collectively reported in Australia since the 2012 to 2014 race seasons. Although information on individual horse fatalities, falls and injuries is published by Racing Victoria in Stewards’ Reports, the data are not aggregated, and so cannot readily be used to assess trends or evaluate the efficacy of safety measures introduced by the industry. The aim of this study was to determine the fatality, fall and injury rates for horses participating in hurdle and steeplechase races in Victoria in the 2022 and 2023 Thoroughbred horse jumps racing seasons compared with horses participating in flat races at the same race meets. Data on horse fatalities, falls and injuries were extracted from the published Racing Victoria race results and Stewards’ Reports for the jumps races (n = 150) and corresponding flat races (n = 157) held at the 38 jumps race meets in Victoria in 2022 and 2023. Overall, horse fatalities, falls and injuries occurred at higher rates in jumps races compared with flat races during the study period. The rate of horse fatalities in jumps races was 3.3 per 1000 starts, with no fatalities in flat races. The rate of horse falls in hurdle races was 24 per 1000 starts and 41.6 per 1000 starts in steeplechase races, comparable with rates previously reported in the 2012 to 2014 seasons. There were no falls in flat races. Horse injuries occurred at a rate of 68.9 per 1000 starts in jumps races compared with 18.8 per 1000 starts in flat races. In hurdle and steeplechase races, veterinary clearance being required following horse injury was 5.4 times (OR 5.4, 95% CI 2.8–10.2) and 7.2 times (OR 7.2, 95% CI 3.3–15.6) more likely, respectively, compared with flat races. The risk of trauma was 4 times more likely in hurdle and steeplechase races (OR 4.8, 95% CI 1.7–13.3 and OR 4.1, 95% CI 1.2–13.4, respectively) and the risk of lameness was increased by 2.5 times in hurdles (OR 2.5, 95% CI 1.2–5.2) and 5.1 times in steeplechase races (OR 5.1, 95% CI 2.3–11.5), compared with flat races. These findings support concerns about the welfare of horses involved in jumps racing and of the need for further safety measures to reduce these risks. Full article

In vitro gamete derivation has been proposed as an interesting strategy for treatment of infertility, improvement of genetic traits, and conservation of endangered animals. Spermatogonial stem cells (SSCs) are primary candidates for in vitro gamete derivation; however, recently, mesenchymal stem cells (MSCs) have also been proposed as candidates for germ cell (GCs) differentiation mainly due to their transdifferentiating capacity. The objective of the present study was to compare the potential for GC differentiation of bovine peripheral blood-derived MSCs (PB-MSCs) and SSCs under the effect of conditioned medium (CM) derived from Sertoli cells (SCs/CM). Samples were collected every 7 days for 21 days and analyzed for pluripotent, GC, and MSC marker expression. The absence of OCT4 and the increased (p < 0.05) expression of NANOG seems to play a role in SSC differentiation, whereas the absence of NANOG and the increased expression (p < 0.05) of OCT4 may be required for PB-MSC differentiation into GCs. SSCs cultured with SCs/CM increased (p < 0.05) the expression of PIWIL2 and DAZL, while PB-MSCs cultured under the same condition only increased (p < 0.05) the expression of DAZL. Overall, the patterns of markers expression suggest that PB-MSCs and SSCs activate different signaling pathways after exposure to SCs/CM and during differentiation into GCs. Full article

In dogs, the spleen is a secondary lymphoid organ that can be affected by both neoplastic and non-neoplastic nodules. In general, few studies relate histopathological diagnosis to tumor size and the number of nodules in spleen biopsies. Some of these studies are inconclusive regarding the difference between neoplastic and non-neoplastic lesions and have small sample sizes or do not consider all splenic lesions. This study aimed to characterize splenic masses and determine risk factors for spleen tumors in dogs. A total of 507 histological reports corresponding to the diagnosis of splenic lesions in dogs from a private laboratory of animal pathology in the Metropolitan Region, Chile, were used. Data were analyzed by descriptive statistics and multiple logistic regression. The most frequent neoplastic and non-neoplastic diagnoses were hemangiosarcoma and hyperplasia, respectively. Most of the cases occurred in male (265 cases, 52.3%), senior (421 cases, 83%), and purebred individuals (342 cases, 67.5%). The most affected breeds were the Cocker Spaniel, German Shepherd, and Labrador Retriever. The most frequent lesion was a single nodule. The variables that exhibited a greater risk for the presentation of splenic neoplasia were male sex (odds ratio (OR) = 16.21; 95% confidence interval (CI) 1.741–150.879; p = 0.014), the presence of two or more splenic nodules (OR = 3.94; 95% CI 2.168–7.177; p < 0.001), an increase in nodule size greater than 2 cm (OR for quartiles 2, 3 and 4 of 2.2; 95% CI 1.036–4.941; p = 0.041, 2.9; 95% CI 1.331–6.576; p = 0.008, and 3.6; 95% CI 1.562–8.499; p = 0.003, respectively), and increasing age (OR = 1.23; 95% CI 1.048–1.436; p = 0.011). On the other hand, males exhibited a lower risk as age increases (OR = 0.76; 95% CI 0.615–0.928; p = 0.008). In conclusion, this study identified that males, multinodular presentation, nodule size, and age are risk factors for the occurrence of splenic neoplasia in dogs, knowledge that will contribute to the diagnostic management of dogs with spleen lesions. Full article

Sheep ectoparasites such as chewing lice, fleas and ticks are serious constraints to sheep productivity and are the cause of skin lesions in animals that decrease their market value. This study aims at investigating the ectoparasite fauna infesting small ruminants in the district of Sidi Bouzid (central Tunisia). A total of 1243 Barbarine and Queue Fine de l’Ouest (QFO) sheep were examined every two months for one year. Of the total animals examined, 74 were infested by at least 1 parasite group (5.95%). Three ectoparasite groups were identified as Psoroptes ovis (0.48%; 6/1243), ticks (5.3%; n = 66/1243) and one specimen of Ctenocephalides canis (0.08%; n = 1/1243). The most abundant tick among the 358 specimens was Rhipicephalus sanguineus sensu lato (n = 337; 94.1%), followed by Hyalomma impeltatum (n = 7/358; 1.9%), H. dromedarii (n = 7/358; 1.9%), H. excavatum (n = 5/358; 1.4%) and only two specimens of H. scupense (n = 2/358; 0.55%). The sheep herds showed low infestation prevalence by ectoparasite over the year, with a significant difference according to the seasons (p < 0.05). A higher infestation prevalence was recorded in March (14.36%). Barbarine sheep breed showed significantly higher infestation prevalence (16.8%) compared to QFO (0.8%) (p < 0.01). There were no differences in infestation prevalence according to sex of the animal or age groups. Knowledge of the ectoparasite population harboured by sheep, its activity dynamics and risk factors is required to develop effective ectoparasite control options. The low prevalence of ectoparasite infestation in sheep reported here may be due to possible genetic resistance or simply to successful hygiene measures implemented by farmers. Full article

Rabbits, pivotal in the EU as livestock, pets, and experimental animals, face bacterial infection challenges, prompting a quest for alternatives to curb antibiotic resistance. Bovine colostrum (BC), rich in immunoregulatory compounds, antimicrobial peptides, and growth factors, is explored for disease treatment and prevention. This study assesses BC diet supplementation effects on rabbit intestines, examining gene expression. Thirty female New Zealand White rabbits at weaning (35 days) were divided into three experimental groups: control (commercial feed), 2.5% BC, and 5% BC. The diets were administered until slaughtering (81 days). BC-upregulated genes in the jejunum included IL-8, TGF-β, and CTNN-β1 at 5% BC, while PLVAP at 2.5% BC. Antioxidant-related genes (SOD1, GSR) were downregulated in the cecum and colon with 2.5% BC. BC 5% promoted IL-8 in the jejunum, fostering inflammation and immune cell migration. It also induced genes regulating inflammatory responses (TGF-β) and gastrointestinal permeability (CTNN-β1). BC 5% enhanced antioxidant activity in the cecum and colon, but no significant impact on anti-myxo antibody production was observed. These results suggest that BC has significant effects on the rabbit gastrointestinal tract’s inflammatory and antioxidant response, but further research is required to fully understand its histological and physiological impact. Full article

(1) Background: The number of horses suffering from chronic respiratory diseases, resembling human asthma, is increasing but there is still a lack of reliable and accurate methods to detect these disorders. Numerous studies have found elevated plasma concentrations of one of the myokines, namely, myostatin (MSTN), in people suffering from severe asthma. MSTN normally inhibits myoblast proliferation and differentiation through autocrine or paracrine signals. Therefore, given the pathogenesis of asthma, we hypothesize that MSTN could be a useful biomarker of equine asthma. Thus, this study aimed to compare the concentration of MSTN in the blood plasma of fully healthy and asthmatic horses. (2) Methods: A total of 61 horses were clinically examined to confirm or exclude the occurrence of equine asthma, including bronchoalveolar lavage (BAL) fluid cytology performed on 49 horses. This study included three groups of horses, two of which were clinically healthy, and one of which was asthmatic. (3) Results: The mean circulatory MSTN concentration determined using the ELISA method in asthmatic horses was significantly higher than that in clinically healthy young Thoroughbred racehorses (p < 0.05), but it did not differ as compared to the group of healthy, adult leisure horses. (4) Conclusions: The obtained results did not unambiguously support our original hypothesis that MSTM may be a reliable marker for the early diagnosis of equine asthma. To the best of the authors’ knowledge, this is the first study to analyze the plasma MSTN concentration in equine asthma patients, and therefore further studies are needed to confirm our novel findings. Full article

Understanding dietary behavior during the individual development of marine predators and its temporal variations elucidates how species adapt to changes in marine resources. This is crucial for predicting marine predators’ habitat selection and the natural population’s responses to environmental changes. The authors conducted a comparative analysis of dietary shift strategies and trophic level variations in Greater lizardfish (Saurida tumbil) in the Beibu Gulf during two distinct periods (2010 and 2020) using stomach content and stable isotope analysis methods. Possible driving factors for these changes were also explored. Changes in the fishery community structure and the decline in the abundance of primary prey resources have led the S. tumbil population to diversify their prey species, utilize alternative resources, and expand their foraging space. However, the species’ foraging strategy, characterized by chasing and preying on schooling and pelagic prey, promoted stability in their feeding behavior across spatial and temporal scales. The main prey items remained demersal and pelagic fish species, followed by cephalopods and crustaceans. Similar to other generalist fish species, ontogenetic dietary shifts (ODSs) indicated a partial transition towards larger prey items. However, the timing and magnitude of the ODSs varied between the two periods, reflecting life-history variations and adaptive adjustments to environmental changes. In comparison to 2010, the population’s mean body length (BL) increased in 2020, and the proportion of the population feeding on pelagic–neritic prey significantly increased. However, the δ¹⁵N values were lower, indicating that the shift in the ecological niche of preferred prey from demersal to pelagic–neritic was the primary cause of the decrease in trophic levels. In the future, we will conduct further quantitative research integrating the spatiotemporal data of both predators and prey to clarify the relationships between marine predators’ feeding behavior, trophic levels, and changes in prey community structure. Full article

While general enrichment strategies for captive animals attempt to elicit variable and species-typical behaviors, approaches to cognitive enrichment have been disappointingly one-size-fits-all. In this commentary, we address the potential benefit of tailoring cognitive enrichment to the “cognitive niche” of the species, with a particular focus on a reasonably well-studied marine carnivore, the sea lion. Sea lions likely share some cognitive evolutionary pressures with primates, including complex social behavior. Their foraging ecology, however, like that of many terrestrial carnivores, is based on the rapid and behaviorally flexible pursuit of avoidant prey. Unlike terrestrial carnivores, sea lions carry out this pursuit in a truly fluid three-dimensional field, computing and executing sensorimotor transformations from any solid angle to any other. The cognitive demands of flexible prey pursuit are unlikely to be fully elicited by typical stationary puzzle box style foraging enrichment devices or screen-based interactive games. With this species, we recommend exploring more water-based movement activities generally, and complex pursuit challenges specifically. Full article

The high-mobility group AT-hook 2(HMGA2) gene has been widely studied in the context of cancer and animal growth. However, recently, several studies have uncovered its critical role in cell proliferation. A genome-wide association study (GWAS) further suggests that the HMGA2 gene is a candidate gene in fertility, indicating its connection not only to growth traits but also to reproduction, specifically ovarian traits. Thus, this study aimed to analyze the distribution of the HMGA2 gene in 54 bovine breeds worldwide, identify important short fragment variants (indels), and investigate the relationship between HMGA2 and ovarian development. The dataset included genotypic information from a bovine population of 634 individuals (n = 634). After genotyping and analyzing four selected loci, we found that one out of four loci, rs133750033 (P4-D_22-bp), was polymorphic. Our results also reveal that this indel of HMGA2 is significantly associated with certain ovarian traits (p < 0.05). Specifically, it has connection with ovarian length (p = 0.004) and ovarian height (p = 0.026) during diestrus. Additionally, we discovered a higher expression of the HMGA2 gene in Asian cattle breeds. In summary, this study suggests that HMGA2 has the potential to serve as an animal fertility testing marker gene. Moreover, these findings contribute to a more promising outlook for the bovine industry. Full article

Canine chronic biliary tree disease (CBTD) is a suspected risk factor for pancreatic injury. The aim of this study was to evaluate the frequency and features of pancreatic involvement in canine CBTD, and their relationship with hyperlipemia and its severity. CBTD was defined as the increase in at least two of ALP, GGT, total bilirubin, cholesterol, and a biliary tree abnormal abdominal ultrasound (graded mild to severe). Pancreatic ultrasound appearance was recorded and classified as acute/chronic. Dogs were divided into a PBD group (pancreatic and biliary disease) and BD group (only biliary tree disease). PBD group was subgraded into a “pancreatic injury” and “pancreatitis” group. Eighty-one dogs were retrospectively included: 56 in the PBD group and 25 in the BD group. Of the PBD group, 20 had pancreatitis (15 chronic and 5 dogs acute). US score was mild in 64 dogs and moderate in 17 dogs, and it was not associated with evidence of pancreopathy. Sixty-six dogs had hyperlipemia (mild = 27 dogs; moderate-to-severe = 39 dogs) and no association with pancreopathy was found. Pancreatic injury was more frequent than pancreatitis in CBTD dogs. Although both acute and chronic pancreatic injury may be present, chronic forms were more frequent. Pancreatic injury should be considered in CBTD patients due its possible clinical significance. Full article

The primary factor leading to elevated rates of diarrhea and decreased performance in piglets is immunological stress. The regulation of immune stress through the intestinal flora is a crucial mechanism to consider. In total, 30 weaned piglets were randomly allocated to five groups: the basal diet group (Control), basal diet + lipopolysaccharides group (LPS), basal diet + 250 μg/kg 6-Formylindolo [3,2-b] carbazole + LPS group (FICZ), basal diet + 3mg/kg Cardamonin + LPS group (LCDN), and basal diet + 6mg/kg Cardamonin + LPS group (HCDN/CDN). The results showed that compared with those of the LPS group, the expression of tight junction proteins (occludin; claudin-1) in the FICZ group was significantly increased, and the mRNA levels of IL-1β and TNF-α were significantly reduced (p < 0.05). HCDN treatment had a better effect on LPS-induced intestinal barrier damage in this group than it did in the LCDN group. HCDN treatment leads to a higher villus height (VH), a higher ratio of villi height to crypt depth (V/C), higher tight junction proteins (ZO-1; occludin), and higher short-chain fatty acids (SCFAs). In addition, correlation analyses showed that Succinivibrio was positively correlated with several SCFAs and negatively correlated with prostaglandin-related derivatives in the FICZ group and CDN group (p < 0.05). In summary, Cardamonin alleviates intestinal mucosal barrier damage and inflammatory responses by regulating the intestinal microbiota and its metabolism. Full article

Most of the previous studies on the genetic variability in Spanish “Berrenda” breeds have been carried out using DNA microsatellites. The present work aimed to estimate the genetic diversity, population structure, and potential genetic differences among individuals of both Berrenda breeds and groups based on the presence of the Robertsonian chromosomal translocation, rob (1;29). A total of 373 samples from animals belonging to the two breeds, including 169 cases diagnosed as rob (1;29)-positive, were genotyped using an SNP50K chip. The genetic diversity at the breed level did not show significant differences, but it was significantly lower in those subpopulations containing the rob (1;29). Runs of homozygosity identified a region of homozygosity on chromosome 6, where the KIT (KIT proto-oncogene, receptor tyrosine kinase) gene, which determines the typical spotted coat pattern in both breeds, is located. The four subpopulations considered showed minor genetic differences. The regions of the genome that most determined the differences between the breeds were observed on chromosomes 4, 6, 18, and 22. The presence of this Robertsonian translocation did not result in sub-structuring within each of the breeds considered. To improve the reproductive performance of Berrenda breeds, it would be necessary to implement strategies considering the involvement of potential breeding stock carrying rob (1;29). Full article

There is still limited information on the genomic structure and genetic diversity of African pigs. Genetic diversity studies can contribute significantly to the genetic improvement and conservation of African pigs. This study presents a genetic diversity analysis and population structure of pig breeds in Ghana, with a focus on the Ashanti Dwarf pig (ADP), an indigenous pig breed of Ghana. A total of 167 pigs sampled in Ghana and populations consisting of Ashanti Dwarf pigs (n = 106), exotics (mostly European pigs) (n = 11), crosses (between indigenous and exotic breeds) (n = 44), and unknown breeds (nondescript) (n = 6) were genotyped using Porcine SNP60K BeadChip. Moderate heterozygosity levels, ranging from 0.28 for Ashanti Dwarf pigs to 0.31 for exotic pigs (mostly European pigs), were observed. Principal component analysis of the pig populations within Ghana resulted in two distinct clusters of pigs: (i) Northern and (ii) Southern regional clusters. The PCA based on breed also resulted in four clusters: (i) ADPs; (ii) exotics (iii) crossbreeds between ADP and exotics; (iv) unknown breed types. The PCA demonstrated that the clustering was influenced by genetics, geographical location, production systems, and practices. ADMIXTURE-based analysis also showed that the populations within Ghana are admixed. FST analysis revealed SNPs associated with QTLs for traits such as disease resilience and growth among ADP populations within the different regional and ecological zones of Ghana. Full article

The fertility rate and litter size of female pigs are critically affected by the expression of estrus. The objective of this study was to elucidate the regulatory mechanisms of estrus expression by analyzing the differential expression of genes and long intergenic non-coding RNAs (lincRNA), as well as the utilization of alternative polyadenylation (APA) sites, in the vulva and vagina during the estrus and diestrus stages of Large White and indigenous Chinese Mi gilts. Our study revealed that the number of differentially expressed genes (DEG) in the vulva was less than that in the vagina, and the DEGs in the vulva were enriched in pathways such as “neural” pathways and steroid hormone responses, including the “Calcium signaling pathway” and “Oxytocin signaling pathway”. The DEGs in the vagina were enriched in the “Metabolic pathways” and “VEGF signaling pathway”. Furthermore, 27 and 21 differentially expressed lincRNAs (DEL), whose target genes were enriched in the “Endocrine resistance” pathway, were identified in the vulva and vagina, respectively. Additionally, we observed that 63 and 618 transcripts of the 3′-untranslated region (3′-UTR) were lengthened during estrus in the vulva and vagina, respectively. Interestingly, the genes undergoing APA events in the vulva exhibited species-specific enrichment in neural or steroid-related pathways, whereas those in the vagina were enriched in apoptosis or autophagy-related pathways. Further bioinformatic analysis of these lengthened 3′-UTRs revealed the presence of multiple miRNAs binding sites and cytoplasmic polyadenylation element (CPE) regulatory aspects. In particular, we identified more than 10 CPEs in the validated lengthened 3′-UTRs of the NFIX, PCNX4, CEP162 and ABHD2 genes using RT-qPCR. These findings demonstrated the involvement of APA and lincRNAs in the regulation of estrus expression in female pigs, providing new insights into the molecular mechanisms underlying estrus expression in pigs. Full article

Marmosets are routinely used in biomedical research, therefore there is an increasing need for updated reference intervals calculated using a large sample size, correct statistics, and considering different variables. Hematological and biochemical values from 472 healthy common marmosets sedated with alphaxalone were collected over a ten-year period (2013–2023). The variables assumed to have influenced the blood-based parameters were compared, i.e., sex, age, housing condition, pregnancy, and contraceptive use. Reference intervals were calculated based on observed percentiles without parametric assumptions, and with parametric assumptions following Box–Cox transformation. Juvenile marmosets showed increased ALP, phosphate, WBC, lymphocyte count, and basophil count and decreased levels of GGT and Fe compared to adults. Marmosets housed strictly indoors showed increased ALT and GGT levels and decreased levels of total bilirubin and neutrophil count compared to marmosets housed with outdoor access. Pregnant marmosets showed increased ALP, total bilirubin, neutrophil count, monocyte count, and basophil count, and decreased levels of AST, ALT, cholesterol, Fe, and lymphocyte count compared to non-pregnant marmosets. Etonogestrel contracepted marmosets showed decreased P-LCR compared to females who were not contracepted. Updated reference intervals will aid researchers and veterinarians in identifying physiological and pathological changes, as well as improve the reproducibility of research in this species. Full article