Animals, Volume 13, Issue 14 (July-2 2023) – 155 articles

Isotopic analysis of carbon and nitrogen in faeces is a reliable methodology for studying ecology in wildlife. Here, we tested this technique to detect variations in carbon and nitrogen isotopic ratios (δ¹³C and δ¹⁵N) in two different intrasexual competition scenarios of male Iberian red deer (Cervus elaphus hispanicus) using faeces of individuals collected during hunting actions in South-eastern Spain. The carbon isotopic ratio (δ¹³C) was not found to be significant, likely due to similar diet composition in all individuals. However, the nitrogen isotopic ratio (δ¹⁵N) was found to be lower in populations where sexual competition between males during the rut was higher compared to low-competition populations. Therefore, this study suggests a different use of proteins by an individual male red deer depending on the sexually competitive context in which he lives. Although further research is needed, these results show the potential of isotopic analysis as a tool for studying individual and populational variations in the level of intrasexual competition, with implications in evolutionary ecology and population management. Full article

The effect of individual and combined supplementation of FA and GPM on physiological variables, productive performance, and carcass characteristics of finishing pigs under heat stress conditions were investigated. Forty Yorkshire × Duroc pigs (80.23 kg) were individually housed and randomly distributed into 4 groups under a 2 × 2 factorial arrangement (n = 10): Control (basal diet, BD); FA, BD + 25 mg FA; GPM, BD with 2.5% GPM; and MIX, BD with 25 mg FA and 2.5% GPM. Additives were supplemented for 31 days. The inclusion of FA or GPM did not modify rectal temperature and respiratory rate. There was an effect of the interaction on FI, which increased when only GPM was supplemented, with respect to Control and MIX (p < 0.05). Average daily gain (ADG) and feed conversion (FC) were not affected by treatments (p > 0.05). The inclusion of FA improved hot and cold carcass weight, while the addition of GPM decreased the marbling (p < 0.05) and tended to increase loin area (p < 0.10). GPM increased liver weight (p < 0.05). The addition of GPM and FA can improve some carcass characteristics under heat stress conditions. It is necessary to continue investigating different levels of inclusion of GPM and FA in finishing pigs’ diets. Full article

Chemotherapy and cyclooxygenase inhibitors (COXi) are primary treatments for canine urethral transitional cell carcinoma (uTCC), a tumor known for its aggressiveness and poor prognosis. This retrospective study aimed to evaluate the clinico-pathological characteristics, treatment modalities, and prognostic factors of 35 dogs with confirmed uTCC that received chemotherapy and COXi. Upon admission, urethral obstruction (UO) and urinary tract infection (UTI) were observed in seven (20%) dogs each. Gemcitabine (n = 20; 57.1%) and vinblastine (n = 10; 28.6%) were commonly used as first-line therapies, with four dogs also receiving radiation therapy. Based on RECIST, one (2.9%) dog achieved complete remission, nine (25.7%) partial remission, 20 (57.14%) showed stable disease, and five (14.3%) progressed. Among dogs with UO, six (85.7%) showed resolution or improvement after the first chemotherapy dose. The median time to local progression was 171 days (range: 107–235), and the median survival time was 333 days (range: 158–508). Dogs with UO upon admission had a higher risk of local progression, while both UO and UTI were associated with an increased risk of overall disease progression and tumor-related death. Additionally, gemcitabine significantly improved metastatic control. This study identified UO and UTI as negative prognostic factors, highlighting the importance of a multimodal approach in managing uTCC. Full article

Analysis of the mtDNA tRNAleu-COII locus is a widely used tool to establish belonging to a particular evolutionary lineage of Apis mellifera L. (lineages A, M, C, O, and Y). In Russia, most of the area was once inhabited by Apis mellifera mellifera from the M evolutionary lineage, but the introduction of bee subspecies from the southern regions of Russia (A. m. caucasica, A. m. carnica) and from abroad (A. m. carnica, A. m. ligustica) led to fragmentation of their native range. In this study, the results of assessing the haplotype number for the tRNAleu-COII locus of mtDNA in Russian Apis mellifera populations were presented. We analyzed 269 colonies from 19 regions of Russia. As a result, two evolutionary lineages were identified: the East European lineage C (26.4%) and the Northwestern European lineage M (73.6%). A total of 29 haplotypes were identified, 8 of them were already reported, and 21 were found to be novel. From the C lineage, haplotypes C1, C2, C2c, C2j, and C3 were predominant. All M lineage samples from Russia belong to the M17 and M4’ haplogroups but have only minor variations in the form of nucleotide substitutions. An analysis of publications devoted to the tRNAleu-COII locus haplotypes, as well as an analysis of the available tRNAleu-COII sequences in GenBank, showed that there is still a problem with the haplotype nomenclature. Full article

The rabbit is a popular animal model for human biomechanical research involving surgery on the hind limb. Mortality is higher in rabbits when undergoing general anesthesia compared to dogs and cats. Moreover, due to their nature as prey animals, rabbits have a tendency to hide signs of pain, making it challenging to detect discomfort at an early stage. Incorporating regional anesthesia into an anesthetic protocol can greatly reduce the requirements for systemic anesthetic and analgesic drugs, thereby minimizing associated side effects. In other species, a block of the sciatic (ScN) and the femoral nerves (FN) is usually applied in patients undergoing hind limb surgery. In phase 1 of this study, the ScN and the FN have been localized and an appropriate approach has been evaluated under sonographic guidance. In phase 2, a mixture of new methylene blue and lidocaine have been administered to the ScN and the FN in 10 cadavers (20 hind limbs). Staining of the nerves was evaluated by dissection. Ultrasonographically, the ScN appeared as a binocular structure surrounded by a hyperechoic rim. The FN appeared as a hypoechoic structure in the dorsal part of the iliopsoas muscle (IPM), becoming hyperechoic/honey-comb-like in the ventral part. Both nerves could be successfully stained in all animals over a median length of 2.3 cm which was considered effective. This technique allows feasible and accurate access to block the ScN and the FN and may lead to successful analgesia in rabbits undergoing hind limb surgery. Full article

The effectiveness of rabbit-sperm cryopreservation is still below average compared to other domestic species. After the sperm cryopreservation process, post-thawing parameters like motility and membrane integrity are significantly compromised. The use of new extender constituents is an approach that can be used to improve the effectiveness of cryopreservation. Accordingly, we used honey (1.25, 2.5, 5, and 10%), coenzyme Q10 (100 and 200 μM), and β-carotene/α-tocopherol (500 μM/620 μM and 250 μM/310 μM) as candidate components for rabbit-sperm extenders during cryopreservation. Ejaculates from commercial adult rabbit bucks (n = 5) were cryopreserved using conventional freezing. Several post-thawing sperm parameters were assessed, including total motility, membrane integrity, viability, nuclear membrane integrity, acrosome reaction, and mitochondrial membrane potential and activation. Additionally, we performed hormonal analyses of the seminal plasma. Moreover, we analyzed the post-thawing levels of a molecular marker of sperm quality, proAKAP4, which was used in rabbits for the first time. Our findings showed that the 2.5% honey supplementation increased the post-thawing sperm motility (13.75 ± 3.75%) compared to the greater concentrations employed. However, the post-thawing motility was negatively affected by the coenzyme Q10 (0%, in both groups) but was not affected by the β-carotene/α-tocopherol supplementation (22 ± 18.15%, and 11.67 ± 10.17%). In conclusion, the cryopreservation protocols of this study did not help to maintain the sperm parameters after thawing. Further studies are required to identify novel protocols to mitigate the damage caused to rabbit sperm during cryopreservation. Full article

Farmers are continually looking for new, reliable, objective, and non-invasive methods for evaluating the conditions of ewes. Live weight (LW) and body condition score (BCS) are used by farmers as a basis to determine the condition of the animal. Body composition is an important aspect of monitoring animal condition. The body composition is the amount of fat, muscle, and bone; knowing the amount of each is important because the information can be used for better strategic management interventions. Experiments were conducted to establish the relationship between body composition and body parameters at key life stages (weaning and pre-mating), using measurements automatically determined by an image processing application for 88 Coopworth ewes. Computerized tomography technology was used to determine the body composition. Multivariate linear regression (MLR), artificial neural network (ANN), and regression tree (RT) statistical analysis methods were used to develop a relationship between the body parameters and the body composition. A subset of data was used to validate the predicted model. The results showed a correlation between fat, muscle, and bone determined by CT and the fat, muscle, and bone weight estimated by the live weight and body parameters calculated using the image processing application, with r² values of 0.90 for fat, 0.72 for muscle, and 0.50 for bone using ANN. From these results, farmers can utilize these measurements to enhance nutritional and management practices. Full article

Dogs’ ability to cooperate with humans is widely acknowledged, but the factors influencing their spontaneous cooperative tendencies are largely unknown. We investigated whether breed function, training experience, and owner-reported social motivation level contribute to spontaneous dog–owner cooperation. Family dogs (N = 100) of three breed groups (non-working dogs, cooperative/independent working breeds) with various training experiences were tested in an ‘out–of–reach’ task with their owners as their partners, who never directly asked for help during the test. We measured dogs’ behaviour along three main components of successful cooperation: paying attention, understanding the problem, and willingness to cooperate. Breed groups had no significant effect on dogs’ behaviour. No factor was associated with the behavioural variables related to not understanding the task. Dogs with high training levels and high social motivation showed more attention-related behaviours and were more likely to help the owner (training level and social motivation were not correlated with each other). Our results highlight the importance of training experience and social motivation in dogs’ attentiveness and spontaneous cooperativity. This also points to the need for careful sample balancing and experimental procedures that do not rely on specific trained skills. Full article

Thromboelastography (TEG) is a viscoelastic technique that allows the examination of both cellular and plasma protein clotting factors. Thromboelastography helps to investigate the underlying coagulopathy and to monitor therapeutic modalities. Although viscoelastic techniques have been used in human and veterinary medicine, reference ranges in pet rabbits are missing. The objective of this study is to establish the reference-range values of TEG parameters in healthy pet rabbits. 24 healthy pet rabbits of different breeds were included: 16 crossbreeds, four Californians, two lops, one lionhead, and one angora. Four rabbits were less than one year old and 20 were older than one year. Twelve rabbits were neutered females, 10 neutered males, and two were intact females. Health status was assessed through a physical examination, a complete blood work, and a coagulation profile. A TEG 5000 Thromboelastograph Hemostasis System was used with kaolin-activated citrated whole blood. All samples were analysed 30 min postextraction. The TEG reference ranges were reaction time (R) 1.4–6.9 min; clot formation time (K) 0.8–2.2 min; α angle 65.8–82.2 degrees; maximal amplitude (MA) 53.7–73.5 mm; measure of clot strength/firmness (G-value) 5796.6–13,885.9 dyn/cm²; and percentage of clot lysis in 30 min (LY30%) 0–41.5%. This study provides the reference ranges of TEG in pet rabbits. Full article

General anesthesia for ovariectomy in dogs is based on a balanced anesthesia protocol such as using analgesics along with an inhalant agent. While opioids such as fentanyl and methadone are commonly used for their analgesic potency, other drugs can also have analgesic effects. Maropitant, an antiemetic for dogs and cats, has also been shown to exert analgesic effects, especially on visceral pain. The aim of this study was to compare the cardiorespiratory effects and analgesic properties of maropitant and methadone combined with desflurane in dogs undergoing ovariectomy. Two groups of 20 healthy mixed-breeds bitches undergoing elective ovariectomy received intravenous either maropitant at antiemetic dose of 1 mg kg⁻¹ or methadone at the dose of 0.3 mg kg⁻¹. Cardiorespiratory variables were collected before premedication, 10 min after sedation and during surgery. Recovery quality and postoperative pain were evaluated 15, 30, 60, 120, 240 and 360 min postoperatively. Results showed that maropitant produced analgesia and reduced the requirement of desflurane in amounts similar to those determined by methadone (5.39 ± 0.20% and 4.91 ± 0.26%, respectively) without significant difference, while maintaining heart rate, arterial blood pressure, respiratory rate and carbon dioxide end-tidal partial pressure even at a more satisfactory level. Therefore, maropitant may be recommended as an analgesic drug for abdominal surgery not only in healthy dogs but also in those with reduced cardiorespiratory compensatory capacities or at risk of hypotension, especially when combined with a sedative such as dexmedetomidine. Full article

Stunning by carbon dioxide (CO₂) inhalation is controversial because it is associated with vigorous movements and behaviours which may or may not be conscious reactions. Furthermore, it is unknown whether some behaviours might indicate the transition into unconsciousness. Our study objective was to investigate the loss of consciousness during CO₂ stunning by linking physiological variables (in particular pH, PaO₂ and PaCO₂) to the onset of observed behaviours. A total of 11 cross-bred pigs were studied. A tracheostomy tube, venous and arterial cannulae were placed under sevoflurane anaesthesia. After recovery from this, and a “wash out” period of at least 30 min, arterial blood samples were taken (and baseline values established) before 90–95% CO₂ in medical air was administered through the tracheostomy tube. Subsequent behaviours were video-recorded and key physiological variables were evaluated using an anaesthetic monitor and the frequent sampling of arterial blood (albeit with inconsistent inter-sample intervals). After the study, behaviours were classified in an ethogram. At the onset of behaviours categorised as “vigorous movement extremities”, “opisthotonos” and “agonal gasping” pH values (range) were: 6.74–7.34; 6.66–6.96 and 6.65–6.87, while PaCO₂ (kPa) was 4.6–42.2, 24.4–51.4 and 29.1–47.6. Based upon these values, we conclude that the pigs were probably unconscious at the onset of “opisthotonos” and “agonal gasping”, but some were probably conscious at the onset of “vigorous movements”. Full article

Footbaths containing disinfectants are used on dairy farms to reduce the spread of digital dermatitis; however, they commonly become contaminated with manure. This trial investigated the physical properties and microbial composition of dairy cow manure from two production systems and examined whether the source of manure impacted the efficacy of footbathing disinfectants. Manure was collected from eighteen dairy cows, nine housed and fed grass silage (HOUSED) and nine at pasture (PASTURE). The pH and dry matter content was determined, total DNA was extracted and the region v3-v4 of the 16s rRNA gene sequenced. The efficacy of formalin and two trial products (TP1: peracetic acid and hydrogen peroxide; TP2: chlorocresol and triamine) was evaluated when mixed with manure from the two production systems. Production system differences were found in manure dry matter content, bacterial microbiome and the efficacy of both trial footbathing products but not formalin. The properties of manure affected the results of laboratory testing and therefore have the potential to influence footbathing disinfectant efficacy when footbaths are contaminated with manure. Further research into the impact of organic contaminants on the efficacy of disinfectants could facilitate the development of improved testing programmes and disinfectant products. Full article

Facial phenotypes are significant in communication with conspecifics among social primates. Less is understood about the impact of such markers in heterospecific encounters. Through behavioral and physical phenotype analyses of domesticated dogs living in human households, this study aims to evaluate the potential impact of superficial facial markings on dogs’ production of human-directed facial expressions. That is, this study explores how facial markings, such as eyebrows, patches, and widow’s peaks, are related to expressivity toward humans. We used the Dog Facial Action Coding System (DogFACS) as an objective measure of expressivity, and we developed an original schematic for a standardized coding of facial patterns and coloration on a sample of more than 100 male and female dogs (N = 103), aged from 6 months to 12 years, representing eight breed groups. The present study found a statistically significant, though weak, correlation between expression rate and facial complexity, with dogs with plainer faces tending to be more expressive (r = −0.326, p ≤ 0.001). Interestingly, for adult dogs, human companions characterized dogs’ rates of facial expressivity with more accuracy for dogs with plainer faces. Especially relevant to interspecies communication and cooperation, within-subject analyses revealed that dogs’ muscle movements were distributed more evenly across their facial regions in a highly social test condition compared to conditions in which they received ambiguous cues from their owners. On the whole, this study provides an original evaluation of how facial features may impact communication in human–dog interactions. Full article

This study examined the anomalous variations of the ventral process of C6 in modern E. ferus caballus. The aim was to provide an incremental grading protocol measuring the absence of the caudal ventral tubercle (CVT) in this ventral process. The findings revealed the most prevalent absent CVT (aCVT) was left unilateral (n = 35), with bilateral (n = 29) and right unilateral (n = 12). Grading was determined in equal increments of absence 1/4, 2/4, 3/4, with 4/4 representing a complete aCVT in 56/76, with a significance of p = 0.0013. This also applied to bilateral specimens. In those C6 osseous specimens displaying a 4/4 grade aCVT, 41/56 had a partial absence of the caudal aspect of the cranial ventral tubercle (CrVT). Here, grading absent CrVTs (aCrVT) followed similarly to aCVTs, though 4/4 was not observed. The significance between 4/4 grade aCVTs and the presentation of an aCrVT was left p = 0.00001 and right p = 0.00018. In bilateral specimens, C6 morphologically resembled C5, implying a homeotic transformation that limited the attachment sites for the cranial and thoracal longus colli muscle. This potentially diminishes function and caudal cervical stability. Therefore, it is recommended that further studies examine the morphological extent of this equine complex vertebral malformation (ECVM) as well as its interrelationships and genetic code/blueprint. Full article

The goat meat preferred by consumers in Spain comes from suckling goat kids, slaughtered at a live weight of 8–10 kg. However, consumption of this meat is very uncommon, so it is necessary to show its value. To achieve this, we planned to investigate consumers’ perceptions about goat kid meat and to study whether their perceptions are related to their sensory appraisal of the meat, measured by the mean of the consumers’ home tests. The experiment was conducted with 30 volunteer families (from two cities with different consumption patterns), who were surveyed regarding their sociodemographic parameters, purchasing and eating habits, and the importance of the meat’s attributes. As a result, four clusters were obtained, which were labeled “negative,” “idealistic,” “positive,” and “concerned about fat content”. The parameters of the animal production system were valued differently by the clusters. Meat tenderness, taste, and amount of fat were considered very decisive factors by most respondents. When the goat kid meat was valued, tenderness was considered more important than taste among older people (“negative” cluster), whereas there was not so much difference between the appraisal of all parameters for the other three clusters. We conclude that it is necessary to improve the information received by consumers about goat production systems and meat quality parameters. There is certainly potential for creating new markets, especially targeted toward young consumers and considering specific strategies for the different groups of consumers, depending on the region and habits of consumption. Full article

There are about 10.1 million domestic cats in Italy, but information on cats’ litter management and house-soiling prevalence is scant. This study described cats’ and cat owners’ profiles, litter management practices, and whether cats show house-soiling, also comparing between professionals (i.e., breeders) and amateurs (i.e., pet owners). A cross-sectional online survey sought respondents’ housing, family, and cat details, as well as other pet details, litter details, and whether the cats showed house-soiling. Data for a total of 3106 cats were obtained. Italian cats lived mainly in apartments, along with other cats or dogs. Italians owned mostly adult European breed cats, to whom they provided covered litter boxes filled with clumping substrates, scooped daily, and completely replaced weekly. Litter cleaning was more frequent when cats were owned for financial purposes (i.e., breeders) rather than for companionship, but more space was provided for pets than for breeding cats. The recalled prevalence of elimination problems (16.7%) was lower compared to other studies, with cats mainly eliminating urine (54.6%) on objects in squatting posture (35.2%). Overall, this research increased our understanding of cat litter management in Italy. These findings could fill a gap in the knowledge regarding litter management and house-soiling incidences in Italy. Further studies to investigate possible risk factors for house-soiling are needed. Full article

In livestock research, there has been a growing interest in the impact of melatonin on both health and disease conditions. The hypothesis of the present study was that melatonin treatment prenatally could support the immune competence and growth of experimentally infected lambs. This is the first study that aimed to investigate the impact of melatonin administration throughout pregnancy on immunity and oocyst excretion of pre-partum ewes and their offspring after experimental infection with Eimeria species. Thirty pregnant ewes were allocated into five equal groups, ΚΜ, ΚC, CM, CC, and NC, and gave birth to 47 lambs. Ewes of the KM and KC groups were orally challenged with a cocktail of Eimeria-sporulated oocysts (mainly consisting of Eimeria ovinoidalis), on day 120 of pregnancy, as well as all the lambs at the age of 5–9 days apart from those born from the NC group (environmental control). Fecal samples were collected from all ewes before infection and at parturition and from all lambs 14 times (S0–S13), before infection and during the following 8 weeks, for counting oocysts per gram of feces (OPG). Immunoglobulin (IgG) and cytokine (IL-1β, IL-6, IL-10, IFN-γ) levels were determined in ewes’ plasma collected before infection and at parturition, in lambs’ plasma at 24 and 72 h after their birth, and in colostrum samples at parturition and 72 h later. Body weight of lambs was recorded five times from birth until the age of 60 days. Accordingly, the leucogram was evaluated in blood samples collected six times within the same period. On average, IgG concentration was higher (p < 0.05) in the blood of KM-ewes compared to KC and CC groups and in colostrum of KM-ewes compared to other groups (p < 0.001). KM-lambs had greater IgG titer and IFN-γ level than the other groups (p < 0.05). The IL-10/ IFN-γ ratio in KM-ewes was lower than the CC group (p = 0.06). Overall, the growth rate of lambs did not differ among groups (p > 0.05). Total oocysts’ excretion in KM- and CM-lambs was reduced by 94.9% (p = 0.05) and 92.6% (p = 0.025), respectively, compared to KC-lambs, following the 3-week period after challenge, when E. ovinoidalis predominated in all groups. The dominant type of leucocytes was monocytes in all experimentally infected lambs, but not in NC-lambs, while overall lymphocytes were lower in KC-lambs than in NC-lambs (p < 0.05). Considering that almost all young indoor-reared lambs are exposed to coccidia species during their early life, melatonin treatment prenatally could suggest an alternative management tool in alleviating infection pressure. Full article

This study aimed to evaluate the effects of dietary Chlorella vulgaris (CV) on the distribution of immune cells, intestinal morphology, intestinal barrier function, antioxidant markers, and the cecal microbiome in 10-day-old broiler chickens. A total of 120 day-old Ross 308 male broiler chicks were assigned to two dietary treatments using a randomized complete block design, with body weight as the blocking factor. Birds fed a diet containing CV showed an increase in CD4⁺ T cells (p < 0.05) compared to those fed the control diet. The relative mRNA expression of intestinal epithelial barrier function-related markers (occludin and avian β-defensin 5) was elevated (p < 0.05) in the CV-supplemented group compared to the control group. The alpha diversity indices (Chao1 and observed features) of the cecal microbiome in 10-day-old birds increased (p < 0.05), indicating higher richness within the cecal bacterial community. In the microbiome analysis, enriched genera abundance of Clostridium ASF356 and Coriobacteriaceae CHKCI002 was observed in birds fed the diet containing CV compared to those fed the control diet. Taken together, dietary CV supplementation might alter intestinal barrier function, immunity, and microbiomes in 10-day-old broiler chickens. Full article

The Kivircik is an indigenous sheep breed from Turkey, and it has superior meat quality compared to other indigenous breeds. Therefore, farmers prioritize Kivircik lamb fattening instead of milk production. Here, we aimed to determine the best nonlinear growth model, i.e., Gompertz, Logistic, Von Bertalanffy, and Brody, to describe the growth curve of Kivircik lambs. The body weight data from birth until 150 days of age belonging to 612 lambs were used as the material of this study. The best fitting model was selected by considering the adjusted coefficient of determination (R²_adj), residual mean square, and Akaike’s (AIC) and Bayesian information criteria (BIC). Even though the Brody model had a better statistical fit, considering its biological interpretation, the Gompertz model was identified as an appropriate model for describing Kivircik lamb growth. Male lambs, twin lambs, and lambs born in winter had higher mature live weights (44.2 kg, 71.2 kg, and 38.5 kg, respectively) and rate of weight gain (2.1, 2.6, and 2.0, respectively). However, our subgroups revealed a similar rate of maturity (0.01). Growth models are important tools for deciding the optimal slaughter age and they provide valuable information on the management practices of both sexes, birth types, and birth seasons. These results can be applied to breeding programs for early selection, enabling intervention strategies when needed. Full article

This study evaluated the effects of supplementing different doses of a multienzyme (KZP) consisting of carbohydrases and a protease on growth performance, duodenal pH and morphology, and carcass traits in broilers fed diets with increasing reductions in energy. One thousand two hundred one-day-old broiler chicks were allocated to five dietary treatments with eight replicates of 30 birds each: a positive control diet formulated to meet Arbor Acres’ nutritional requirements (PC); a negative control diet reformulated to 80 kcal/kg less than the apparent metabolizable energy (AME) of the PC (NC1); a negative control diet reformulated to 120 kcal/kg less than the AME of the PC (NC2); an NC1 diet supplemented with 300 g/t of KZP (NC1 + KZP300); and an NC2 supplemented with 500 g/t of KZP (NC2 + KZP500). Growth performance was measured throughout the study. At 35 days, 10 birds per treatment were randomly selected and euthanized for a carcass trait evaluation, and samples of the duodenum were collected for morphological examination and pH level determination. The final average body weight and feed conversion ratio were better (p < 0.05) for the broilers in the NC1 + KZP300 group compared to those in NC1, NC2 and NC2 + KZP500 groups and were similar to those of the PC birds (p > 0.05). Birds from the NC1 + KZP500 group showed a better (p < 0.05) final body weight and feed efficiency compared to the NC1 and NC2 groups. The villus height was greater (p < 0.05) for the PC and NC1 + KZP300 groups compared to the rest of the treatments. The crypt depth was longer (p < 0.05) for the NC1 and NC2 groups compared to the NC1 + KZP300 group. The supplementation of KZP to both the NC1 and NC2 diets reduced (p < 0.05) the abdominal fat %. This study demonstrates that supplementing energy-reduced diets with KZP improved performance in broiler chickens. Full article

We dissected carcasses of eight mature females, both parous and non-parous specimens, to study the macroscopic anatomy of the female reproductive system in the sugar glider. The genital system includes double organs, namely the right and left ones, which are completely separated. It includes two ovaries, two oviducts, two uteri and a vaginal complex. The uteri are fusiform-shaped and lack horns. The vaginal complex includes two lateral vaginae and a median vagina, also called the ‘birth canal’. The cranial end of both lateral vaginae partially fuses, forming an expansion named the vaginal sinus, which is divided into two parts by a longitudinal septum, one for each vagina, where the ipsilateral uterine cervix opens. The caudal end of the lateral vaginae opens into a medial and impar duct: the urogenital sinus that serves as a common passage for the reproductive and urinary systems. In non-pregnant females, only the lateral vaginae are present. In pregnant and recently parous females, a short median vagina extends from the caudal wall of the vaginal sinus to the cranial end of the urogenital sinus. In the ventral wall of this sinus, next to its caudal opening, there is a forked clitoris. Full article

Pseudorabies virus (PRV) belongs to the family Herpesviridae. PRV has a wide host range and can cause cytopathic effects (CPEs) in PK-15 cells. Therefore, PRV was used as a model to study the antiviral activity of piceatannol. The results showed that piceatannol could restrain PRV multiplication in PK-15 cells in a dose-dependent manner. The 50% inhibitory concentration (IC₅₀) was 0.0307 mg/mL, and the selectivity index (SI, CC₅₀/IC₅₀) was 3.68. Piceatannol could exert an anti-PRV effect by reducing the transcription level of viral genes, inhibiting PRV-induced apoptosis and elevating the levels of IL-4, TNF-α and IFN-γ in the serum of mice. Animal experiments showed that piceatannol could delay the onset of disease, reduce the viral load in the brain and kidney and reduce the pathological changes in the tissues and organs of the mice to improve the survival rate of the mice (14.3%). Therefore, the anti-PRV activity of piceatannol in vivo and in vitro was systematically evaluated in this study to provide scientific data for developing a new alternative measure for controlling PRV infection. Full article

No system nor standardized analytical procedures at commercial laboratories exist to facilitate and accurately measure potential viable virus contamination in feed ingredients and complete feeds globally. As a result, there is high uncertainty of the extent of swine virus contamination in global feed supply chains. Many knowledge gaps need to be addressed to improve our ability to prevent virus contamination and transmission in swine feed. This review summarizes the current state of knowledge involving: (1) the need for biosecurity protocols to identify production, processing, storage, and transportation conditions that may cause virus contamination of feed ingredients and complete feed; (2) challenges of measuring virus inactivation; (3) virus survival in feed ingredients during transportation and storage; (4) minimum infectious doses; (5) differences between using a food safety objective versus a performance objective as potential approaches for risk assessment in swine feed; (6) swine virus inactivation from thermal and irradiation processes, and chemical mitigants in feed ingredients and complete feed; (7) efficacy of virus decontamination strategies in feed mills; (8) benefits of functional ingredients, nutrients, and commercial feed additives in pig diets during a viral health challenge; and (9) considerations for improved risk assessment models of virus contamination in feed supply chains. Full article

The purpose of this study was to look at research trends in the application of CTM in animal nutrition in order to identify current and emerging challenges, as well as to examine the intellectual structure of the subject. The intellectual structure of CTM was examined using keyword and reference analysis. The research community includes all research and review articles published in journals indexed in the Web of Science database during the years 1990–2022. The results showed that the terms zinc, co-occurring 331 times, performance (324 times), and copper 216 (times) were the main and hotspots of research in the field of chelate. The data suggest that the most important keywords during the study period were zinc, copper, pig, bovine, metabolism, and bioavailability. The terms health, muscle, beef, trace elements, and dietary supplements represent emerging topics in CTM, as research began to focus on these areas during the years 2017–2022. The country with the greatest number of published articles was the United States of America. This bibliometric analysis showed that countries are focusing on the effects of CTM on the health and musculature of cattle through dietary supplementation with trace elements. According to the identified hot and emerging topics, this research can serve as a roadmap for a global comprehensive scientific plan and policy. Full article

Cattle grazing tall fescue (Schedonorus arundinaceous) infected with wild-type endophytes (WE) leads to a syndrome commonly known as fescue toxicosis. Replacing WE tall fescue with a novel endophyte-infected (NE) tall fescue can mitigate this problem but adoption of this technology has been limited. This study measured and determined the physiological and behavioral responses of heifers that grazed either WE or NE tall fescue, utilizing relatively non-invasive techniques including hair cortisol, thermography (for extremity temperatures), small loggers for intravaginal temperature, and remote observation of in-field behavior. Heifers that grazed WE had greater (p < 0.0001) hair cortisol levels, lower extremity temperatures (p ≤ 0.0075), and 0.3–0.9 °C greater (p ≤ 0.02) intravaginal temperatures (particularly during the daytime) than heifers that grazed NE. From 1200 h–1700 h each day, heifers on WE pastures spent 1.5 more (p = 0.0003) hours standing up and 0.9 fewer (p = 0.0402) hours lying down than heifers on NE pastures. Differences (p = 0.0160) in ADG were small (0.1 kg d⁻¹) and were only observed in the first year of these 8-week studies. However, even in the mild environment of the study site, grazing NE tall fescue provided clear welfare benefits as evidenced by heifer behavioral changes, temperature differentials, and hair cortisol levels. This study underscores the potential utility of non-invasive techniques, such as thermographic imaging and hair cortisol analysis, for evaluating animal responses to stress in extensive grazing systems. Full article

In Europe, regulations contain guidance to maintain high standards of animal welfare. In many parts of the world, large buyers for supermarkets or restaurants are the main enforcers of basic animal welfare standards. They can have considerable influence on improving standards on large commercial farms. Research clearly shows that straw is one of the most effective environmental enrichment for pigs. On some large farms, there are concerns that straw will either clog waste management systems or bring in disease. This paper contains a review of both scientific research and practical experience with enrichment devices that are easy to implement. Pigs prefer enrichment objects that they can chew up and deform. Broiler chickens prefer to climb up on objects, hide under them or peck them. It is always essential to uphold basic welfare standards such as animal cleanliness and low levels of lameness (difficulty walking). It is also important to reduce lesions, and maintain body conditions of breeding animals. An environment enrichment device is never a substitute for poorly managed facilities. It should enhance animal welfare on well-managed farms. Full article

Gentile di Puglia (GdP) is an autochthonous sheep breed of Southern Italy included among ovine breeds threatened by genetic erosion and extinction risk, which have been given attention by local and international institutions, thus emphasizing the need for germplasm conservation actions. In the present study, two assisted reproduction approaches, finalized for GdP conservation, were performed: (1) on-farm reproductive efficiency evaluation, expressed as pregnancy rate (PR), twin pregnancy rate (tPR), and body condition score (BCS), for three consecutive breeding cycles and (2) pre-pubertal lambs’ immature cumulus–oocyte complex (COC) retrieval, vitrification, in vitro maturation (IVM), and assessment of meiotic stage and bioenergetic-oxidative status compared with those of other Italian and European commercial breeds. PR and tPR were progressively reduced over time. In all clinical examination times, BCS was significantly lower in nonpregnant ewes compared with pregnant ones. Fresh GdP pre-pubertal lamb COCs achieved meiotic maturation and showed healthy bioenergetic–oxidative status after IVM. Vitrification reduced the oocyte maturation rate in all groups. However, mature oocytes retained their cytoplasmic maturity, expressed as a mitochondria distribution pattern and activity, indicating promising developmental competence. In conclusion, clinical- and biotechnological-assisted reproduction approaches can support conservation strategies of GdP and other local sheep breeds in Southern Italy. Full article

Postpartum negative energy balance (NEB) is detrimental to cows and decreases profitability in dairy farming. The two origins of milk fatty acids (FA), de novo synthesized in the mammary gland and plasma lipids initially originating from feed, rumen microbes and the animal’s adipose tissue, make milk FA candidates as possible NEB biomarkers. The aim of this study was to assess the possibility to predict EB in cows in the first 150 days of lactation with BCS, milk traits and selected individual milk FA and the ratios of blood-derived and de novo synthesized FA. The daily EB of Estonian Holstein cows (N = 30) was calculated based on body weights and BCS values. Milk FA were analyzed with gas chromatography. The variance partitioning analysis revealed that milk production traits, BCS at calving, FA ratios and days in milk accounted for 67.1% of the EB variance. Random forest analysis indicated the highest impact of the ratios C18:1cis9/C12:0+C14:0, C18:1cis9+C18:0/C12:0+C14:0, C18:1cis9/C14:0, C18:1cis9+C18:0/C14:0, C18:1cis9/sum C5:0 to C14:0, C18:1cis9+C18:0/sum C5:0 to C14:0 or C18:1cis9/C15:0. FA and their ratios alone explained 63.6% of the EB variance, indicating the possibility to use milk FA and their ratios as sole predictors for the energy status in dairy cows. Full article

European sea bass production has increased in recent decades. This increase is associated with an annually rising demand for sea bass, which encourages the aquaculture industries to increase their production to meet that demand. However, this intensification has repercussions on the animals, causing stress that is usually accompanied by dysbiosis, low feed-conversion rates, and immunodepression, among other factors. Therefore, the appearance of pathogenic diseases is common in these industries after immunodepression. Seeking to enhance animal welfare, researchers have focused on alternative approaches such as probiotic application. The use of probiotics in European sea bass production is presented as an ecological, safe, and viable alternative in addition to enhancing different host parameters such as growth performance, feed utilization, immunity, disease resistance, and fish survival against different pathogens through inclusion in fish diets through vectors and/or in water columns. Accordingly, the aim of this review is to present recent research findings on the application of probiotics in European sea bass aquaculture and their effect on growth performance, microbial diversity, enzyme production, immunity, disease resistance, and survival in order to help future research. Full article

Sperm proteins play vital roles in improving sperm freezing resilience in domestic animals. However, it remains poorly defined which proteins regulate the freezing resilience of spermatozoa in rams (Ovis aries). Here, we compared the proteome of ram sperm with a high cryopreservation recovery ratio (HCR) with that of ram sperm with a low cryopreservation recovery ratio (LCR) using a tandem mass tag-based quantitative proteomics approach. Bioinformatic analysis was performed to evaluate differentially expressed proteins (DEPs). A total of 2464 proteins were identified, and 184 DEPs were screened. Seventy-two proteins were higher in the LCR group. One hundred and twelve proteins were more abundant in the HCR group, and they were mainly involved in the regulation of oxidative phosphorylation and thermogenesis pathways. Proteins in high abundance in the HCR group included the S100A family, such as S100A8, S100A9, S100A14, and S100A16, effectively controlling for CA²⁺ and maintaining flagella structure; HYOU1 and PRDX1, which participate in antioxidant protection and anti-apoptosis to prevent cell death; and HSP90B1, which maintains cell activity and immune response. Our results could help illuminate the molecular mechanisms underlying cryopreservation of ram semen and expand the potential direction of cryopreservation of high-quality semen. Full article