Animals, Volume 12, Issue 13 (July-1 2022) – 141 articles

On 21 December 2020, the European Commission granted conditional marketing authorisation for the BNT162b2 COVID-19 vaccine ‘Comirnaty’, produced by Pfizer/BioNTech. This happened only twelve months after scientists first identified SARS-CoV-2. This stands in stark contrast with the usual ten years needed for vaccine development and approval. Many have suggested that the changes in required animal tests have sped up the development of Comirnaty and other vaccine candidates. However, few have provided an overview of the changes made and interviewed stakeholders on the potential of the pandemic’s pressure to achieve a lasting impact. Our research question is: how have stakeholders, including regulatory agencies and pharmaceutical companies, dealt with requirements concerning in vivo animal models in the expedited approval of vaccine candidates such as ‘Comirnaty’? We interviewed key stakeholders at the Dutch national and European levels (n = 11 individuals representing five stakeholder groups in eight interviews and two written statements) and analysed relevant publications, policy documents and other grey literature (n = 171 documents). Interviewees observed significant changes in regulatory procedures and requirements for the ‘Comirnaty’ vaccine compared to vaccine approval in non-pandemic circumstances. Specifically, the European Medicines Agency (EMA) actively promoted changes by using an accelerated assessment and rolling review procedure for fast conditional marketing authorisation, requiring a reduced number of animal studies and accepting more alternatives, allowing pre-clinical in vivo animal experiments to run in parallel with clinical trials and allowing re-use of historical data from earlier vaccine research. Pharmaceutical companies, in turn, actively anticipated these changes and contributed data from non-animal alternative sources for the development phase. After approval, they could also use in vitro methods only for all batch releases due to the thorough characterisation of the mRNA vaccine. Pharmaceutical companies were optimistic about future change because of societal concerns surrounding the use of animals, adding that, in their view, non-animal alternatives generally obtain faster, better, and cheaper results. Regulators we interviewed were more hesitant to permanently implement these changes as they feared backlash regarding safety issues and uncertainty surrounding adverse effects. Our analysis shows how the EMA shortened its approval timeline in times of crisis by reducing the number of requested animal studies and promoting alternative methods. It also highlights the readiness of pharmaceutical companies to contribute to these changes. More research is warranted to investigate these promising possibilities toward further replacement in science and regulations, contributing to faster vaccine development. Full article

Bovine leukemia virus (BLV) is a retrovirus that affects cattle worldwide. A longitudinal study was conducted with the aim to (a) estimate the incidence rate of the BLV infection of dairy farms in the regions of Los Ríos and Los Lagos (Chile), and (b) describe the frequency and epidemiological association of risk management practices related to new cases in cattle on dairy farms in Southern Chile. Infection status was based on commercial blocking ELISA results, on serum and milk. Individual information on animals and management practices was extracted from farm records, and then the most likely date of infection for new cases was estimated. The number of new infections was used to calculate the within-herd incidence rate. Adult animals had an incidence rate of 1.16 (95% CI 0.96; 1.20) cases per 100 cow-months at risk, while for young animals it was 0.64 (95% CI 0.44; 1.00) cases per 100 animal-months at risk. Rectal palpation, artificial insemination, and injections were the most common practices related to infection. Further studies are needed to determine if these are the only practices that facilitate spreading or if there are other practices that can be handled better in order to reduce the spread of BLV. Full article

Salmonid species demonstrate varied susceptibility to the viral pathogen infectious hematopoietic necrosis virus (IHNV). In California conservation hatcheries, juvenile Chinook salmon (Oncorhynchus tshawytscha) have experienced disease outbreaks due to L genogroup IHNV since the 1940s, while indigenous steelhead (anadromous O. mykiss) appear relatively resistant. To characterize factors contributing to the losses of California salmonid fish due to IHNV, three populations of Chinook salmon and two populations of steelhead native to California watersheds were compared in controlled waterborne challenges with California L genogroup IHNV isolates at viral doses of 10⁴–10⁶ pfu mL⁻¹. Chinook salmon fry were moderately to highly susceptible (CPM = 47–87%) when exposed to subgroup LI and LII IHNV. Susceptibility to mortality decreased with increasing age and also with a higher temperature. Mortality for steelhead fry exposed to two IHNV isolates was low (CPM = 1.3–33%). There was little intraspecies variation in susceptibility among populations of Chinook salmon and no differences in virulence between viruses strains. Viral persistence was demonstrated by the isolation of low levels of infectious IHNV from the skin of two juvenile Chinook salmon at 215 d post exposure. The persistence of the virus among Chinook salmon used for stocking into Lake Oroville may be an explanation for the severe epidemics of IHN at the Feather River hatchery in 1998–2002. Full article

Earlier springs in temperate regions since the 1980s, attributed to climate change, are thought to influence the earlier arrival of long-distance migrant passerines. However, this migration was initiated weeks earlier in Africa, where the Southern Oscillation, Indian Ocean Dipole, North Atlantic Oscillation drive climatic variability, and may additionally influence the migrants. Multiple regressions investigated whether 15 indices of climate in Africa and Europe explained the variability in timing of arrival for seven trans-Saharan migrants. Our response variable was Annual Anomaly (AA), derived from standardized mistnetting from 1982–2021 at Bukowo, Polish Baltic Sea. For each species, the best models explained a considerable part of the annual variation in the timing of spring’s arrival by two to seven climate variables. For five species, the models included variables related to temperature or precipitation in the Sahel. Similarly, the models included variables related to the North Atlantic Oscillation (for four species), Indian Ocean Dipole (three), and Southern Oscillation (three). All included the Scandinavian Pattern in the previous summer. Our conclusion is that climate variables operating on long-distance migrants in the areas where they are present in the preceding year drive the phenological variation of spring migration. These results have implications for our understanding of carry-over effects. Full article

The aim of this research was to determine if the division between the cervical, dorsal, and croup profiles (three regions commonly assessed for descriptive profile purposes) has a modular basis as well as a morphological integration. For this, a total of 135 digital photographs were obtained, in a lateral view, of adult horses (14 females and 121 geldings; age range: 2–20 years), of the Araucanian breed, an equine population typical of the flooded savannah of Arauca, NE Colombia. From each image, 25 reference points (semi-landmarks) were obtained at the dorsal level of the neck, back, and croup. The hypothesis of the modularity of different body profiles based on differentiated regions was tested using the RV coefficient, and an analysis of two blocks of partial least-squares allowed the evaluation of the level of morphological integration. The results showed that each alloidic group reflected high integration but low modularity. The covariation between the modules was centered mainly on the withers, the loin, and the croup. For the studied profile blocks, no module can be considered. The absence of the fragmentation of the alloidic sets would promote the adaptive capacity of the breed by linking coordinated functional responses to similar selection pressures, e.g., field work. Although the integration between the neck, back, and croup profiles was proven, their modular covariation was low. Full article

White spot syndrome virus (WSSV) has been reported to cause severe economic loss in the shrimp industry. With WSSV being a large virus still under investigation, the 3D structure of its assembly remains unclear. The current study was planned to clarify the 3D structures of WSSV infections in the cell nucleus of red swamp crayfish (Procambarus clarkii). The samples from various tissues were prepared on the seventh day post-infection. The serial sections of the intestinal tissue were obtained for electron tomography after the ultrastructural screening. After 3D reconstruction, the WSSV-associated structures were further visualized, and the expressions of viral proteins were confirmed with immuno-gold labeling. While the pairs of sheet-like structures with unknown functions were observed in the nucleus, the immature virions could be recognized by the core units of nucleocapsids on a piece of the envelope. The maturation of the particle could include the elongation of core units and the filling of empty nucleocapsids with electron-dense materials. Our observations may bring to light a possible order of WSSV maturation in the cell nucleus of the crayfish, while more investigations remain necessary to visualize the detailed viral–host interactions. Full article

The present study was conducted to investigate the effects of glutamine (Gln) supplementation on intestinal inflammatory reaction and mucosa barrier of broilers administrated with lipopolysaccharide (LPS) stimuli. A total of 120 1-d-old male broilers were randomly divided into four treatments in a 2 × 2 experimental arrangement, containing immune challenge (injected with LPS in a dose of 0 or 500 μg/kg of body weight) and dietary treatments (supplemented with 1.22% alanine or 1% Gln). The results showed that growth performance of broilers intra-abdominally injected with LPS was impaired, and Gln administration alleviated the adverse effects on growth performance induced by LPS challenge. Furthermore, Gln supplementation reduced the increased concentration of circulating tumor necrosis factor-α, interleukin-6 and interleukin-1β induced by LPS challenge. Meanwhile, D-lactic acid and diamine oxidase concentration in plasma were also decreased by Gln supplementation. In addition, the shorter villus height, deeper crypt depth and the lower ratio of villus height to crypt depth of duodenum, jejunum and ileum induced by LPS stimulation were reversed by Gln supplementation. Gln administration beneficially increased LPS-induced reduction in the expression of intestine tight junction proteins such as zonula occludens protein 1 (ZO-1), claudin-1 and occludin except for the ZO-1 in duodenum and occludin in ileum. Moreover, Gln supplementation downregulated the mRNA expression of toll-like receptor 4, focal adhesion kinase, myeloid differentiation factor 88 and IL-1R-associated kinase 4 in TLR4/FAK/MyD88 signaling pathway. Therefore, it can be concluded that Gln administration could attenuate LPS-induced inflammatory responses and improve intestinal barrier damage of LPS-challenged broilers. Full article

Background: Owners often feel the cost of veterinary care is too high, as there remains a limited understanding of the cost of health care in human and veterinary medicine alike. Pet health insurance is often seen as a universal solution. However, especially for patient owners with few financial resources, both the bill at the vet and the monthly premium for pet health insurance can become a challenge. Hypothesis: Pet health insurance can prevent or ease many price discussions at the vet, but it does not offer a solution for patient owners with little financial means. Methods: In order to verify for which patient owners pet health insurance can be a solution, four theoretical groups were formed depending on the patient owner’s willingness to pay and his/her dispensable funds based on a theoretical model. Results: Dispensable funds are a factor that cannot be influenced by the veterinary surgeon. However, low dispensable funds as a result of an insufficient willingness to save (whether due to a lack of financial education or a lack of will) can be solved by pet health insurance. Willingness to pay, on the other hand, can be influenced by empathetic communication from the veterinary surgeon and thus also from pet health insurance. Nevertheless, situations remain where pet health insurance is not a solution either, because owners can neither afford the veterinary costs nor a premium for a pet health insurance. Full article

Although there is evidence that pets may help individuals facing significant daily stressors, and that they may enhance the well-being of their owners, little is known about the benefits of pets for job performance. Since the COVID-19 pandemic crisis, teleworking was a strategy implemented in many countries to reduce the virus widespread and to assure organizational productivity. Those who work from home and who own pets may work close to them. Based on the conservation of resources theory, this study aimed to analyze whether positive affect mediated the relationship between telecommuting and self-reported job performance and if psychological and physical closeness to the pet would moderate this relationship in such a way that it would be stronger for those who worked closer to their pet, and who were more emotionally attached to them. For this study, we collected data from 81 teleworkers who did not own pets, and from 320 teleworkers who owned pets. Both answered an online questionnaire. Findings: Results from the study showed the existence of significant differences between those who owned and who did not own pets regarding positive affect and performance, in which those who owned pets reported higher levels of positive affect and self-reported performance and perceived telework more positively. Moreover, positive affect mediated the relationship between telework and self-reported job performance. Furthermore, emotional and physical closeness moderated the mediating effect. This study contributes to a better understanding of the human-animal interaction and how pets can be a personal resource able to change their owners‘ affective experiences and job performance while they are working from home. The findings demonstrate that telework may be a suitable organizational strategy for pet-owners. Full article

Frozen boar sperm is used on a minimal scale in consequence of the cryo-injuries induced by biochemical and physical modifications during the freezing and thawing processes. The present study investigates whether the addition of carboxylated ε-poly-L-lysine (CPLL) to the freezing medium could improve post-thaw boar sperm quality or not. Boar sperm was diluted with freezing medium contained different doses of carboxylated ε-poly-L-lysine (0, 0.125%, 0.25%, 0.5%, and 1%; v/v). The motility patterns, membrane integrity, acrosome integrity, mitochondrial membrane potential, NADH-CoQ activity, ATP level, malondialdehyde (MDA) level, and antioxidant defense system, as well as apoptosis in post-thaw boar sperm, were measured. It was observed that 0.25% CPLL treatment significantly improved the post-thaw boar sperm total motility, progressive motility, straight-linear velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), linearity (LIN), straightness (STR), membrane integrity, and acrosome integrity. Interestingly, the addition of CPLL also significantly increased the post-thaw sperm mitochondrial membrane potential, NADH-CoQ activity, and ATP level. Moreover, post-thaw boar sperm catalase (CAT) activity, glutathione peroxidase (GPx) activity, and superoxide dismutase (SOD) activity were increased with the addition of CPLL from 0.125% to 0.5% concentration levels. Furthermore, reduction of post-thaw sperm MDA level and apoptosis in 0.25% CPLL treatment was also observed. Those observations suggested that the addition of 0.25% CPLL to the freezing medium increased post-thaw boar sperm quality by protecting sperm mitochondrial function and antioxidant defense system. These findings provided novel insights that CPLL can be used as an efficient cryoprotectant to improve the post-thaw boar sperm quality during cryopreservation. Full article

An understanding of fish movement behavior in response to flow field variables is important for exploring the hydrodynamic strategies of fish in fish passages. In this paper, bighead carps were taken as an example. The fish movement behavior response to water flow field information by means of estimating the energetic expenditure using an IBM approach in an experimental fishway was investigated. Fish swimming velocity, drag force, and energy expenditure were analyzed in varied flow conditions related to hydraulic variables, including velocity (V), turbulent kinetic energy (TKE), and strain rate (SR). The result indicated that the fish will require more energy in high TKE zones. This study provides a reference for optimizing the design of fish passages and fisheries management. This method can be applied to assess the efficiency of fish bypass structures and conduct fish survival studies. Full article

The last decade was marked by a steep rise in avian studies at genomic and cellular levels. Cell lines are important tools for in vitro studies in cell biology and cytogenetics. We developed a simple method of primary somatic cell culture establishment from the ovaries of the great tits (Parus major) and testes of ten Passerine species, characterized the cellular composition of the ovary-derived lines using RT-PCR and immunolocalization of the tissue-specific markers and tested the efficiency of two methods of genetic transformation of the ovary-derived cell line. We found that the ovary-derived cell cultures of the great tit were composed of fibroblasts mainly, but also contained interstitial and granulosa cells. They were cultivated until the 10th passage without any noticeable decrease in their proliferative activity. The testis-derived cell cultures had lower proliferative potential. However, both ovary- and testis-derived cell cultures provided enough material for high quality mitotic metaphase chromosome preparations. The efficiency of its transduction with lentivirus containing a GFP reporter was very low, while electroporation with episomal vectors expressing GFP resulted in a high yield of GFP-positive cells. The proposed method could be used for the generation of high quality material for various cytogenetic and genomic studies. Full article

This study examined the protective effect of earthworm extract (EE) on acrylamide (ACR)-induced reproductive dysfunction. Forty male rats were allocated into four groups (n = 10). The G I (control) group received distilled water (D.W.). The G II group received ACR (5 mg kg⁻¹ B.W. in D.W.) 5 days per week, orally, for 3 weeks. The G III group was administered EE (300 mg kg⁻¹ B.W in D.W.) 5 days per week, orally, for 3 weeks. The G IV group was pretreated with EE for 3 weeks and then co-treated with EE and ACR for an additional 3 weeks. ACR decreased the number of sperm, sperm viability, and total motility. However, it increased testosterone levels with no effect on the FSH or LH levels. Moreover, ACR increased the concentrations of malondialdehyde (MDA) and nitric oxide (NO). Meanwhile, it decreased the glutathione (GSH) concentration in testicular tissues. Notably, the expression levels of p53 and Ki-67 were increased in the degenerated spermatogenic cells and in the hyperplastic Leydig cells of the testis of the ACR-treated group, respectively. Acrylamide induced alterations in the testicular tissue architecture. Interestingly, EE restored the sperm parameters and recovered the testicular histological structures and the biochemical alterations induced by ACR. In conclusion, earthworm extract ameliorated ACR-induced reproductive toxicity via restoring the testicular antioxidant balance and suppressing p53 and Ki-67 expressions in testicular tissues. Full article

Osteoclasts are unique and vital bone cells involved in bone turnover. These cells are active throughout the individual’s life and play an intricate role in growth and remodeling. However, extra-label bisphosphonate use may impair osteoclast function, which could result in skeletal microdamage and impaired healing without commonly associated pain, affecting bone remodeling, fracture healing, and growth. These effects could be heightened when administered to growing and exercising animals. Bisphosphonates (BPs) are unevenly distributed in the skeleton; blood supply and bone turnover rate determine BPs uptake in bone. Currently, there is a critical gap in scientific knowledge surrounding the biological impacts of BP use in exercising animals under two years old. This may have significant welfare ramifications for growing and exercising equids. Therefore, future research should investigate the effects of these drugs on skeletally immature horses. Full article

Environmental filtering, spatial factors and species interactions are fundamental ecological mechanisms for community organisation, yet the role of such interactions across different environmental and spatial settings remains mostly unknown. In this study, we investigated fish community organisation scenarios and seasonal species-to-species associations potentially reflecting biotic associations along the Qiupu River (China). Based on a latent variable approach and a tree-based method, we compared the relative contribution of the abiotic environment, spatial covariates and potential species associations for variation in the community structure, and assessed whether different assembly scenarios were modulated by concomitant changes in the interaction network structure of fish communities across seasons. We found that potential species associations might have been underestimated in community-based assessments of stream fish. Omnivore species, since they have more associations with other species, were found to be key components sustaining fish interaction networks across different stream orders. Hence, we suggest that species interactions, such as predation and competition, likely played a key role in community structure. For instance, indices accounting for network structure, such as connectance and nestedness, were strongly correlated with the unexplained residuals from our latent variable approach, thereby re-emphasising that biotic signals, potentially reflecting species interactions, may be of primary importance in determining stream fish communities across seasons. Overall, our findings indicate that interaction network structures are a powerful tool to reflect the contribution of potential species associations to community assembly. From an applied perspective, this study should encourage freshwater ecologists to empirically capture and manage biotic constraints in stream ecosystems across different geographical and environmental settings, especially in the context of the ever-increasing impacts of human-induced local extinction debts and species invasions. Full article

The aim of the study was to test if feeding 15% U. lactuca to broilers, alone or combined with carbohydrases, enhanced meat nutritional quality, without compromising growth performance. One hundred and twenty 22-day-old broilers were allocated to the following diets and replicated 10 times for 14 days: (1) maize and soy-based diet (control); (2) control with 15% U. lactuca (UL); (3) UL diet with 0.005% commercial carbohydrase mixture (ULC); and (4) UL diet with 0.01% ulvan lyase (ULE). Final body weight and average daily gain decreased (p < 0.050) with the ULE diet compared with the control, but no significant differences were found for the other diets. The intestinal viscosity increased (p < 0.001) with all alga diets but was lowered (p < 0.050) in the ileum with the ULE diet, relative to UL and ULC diets. Meat lightness and redness values, off-flavours, and total carotenoids increased (p < 0.001), while yellow values, tenderness, juiciness, overall acceptability, α- and γ-tocopherol, and total lipids decreased (p < 0.001) with alga diets. The n-3 polyunsaturated fatty acids (PUFA) increased (p < 0.050), and the n-6/n-3 PUFA ratio decreased (p < 0.001) with the ULE diet. Total minerals in meat increased (p < 0.001) with alga diets, conversely to sodium and zinc (p < 0.001). Feeding 15% of U. lactuca to broilers did not impair growth but increased meat nutritional value through the accumulation of health-promoting antioxidant carotenoids, n-3 PUFA and total minerals, although reducing overall meat acceptability. Full article

In the last decades, wildlife diseases and the health status of animal populations have gained increasing attention from the scientific community as part of a One Health framework. Furthermore, the need for non-invasive sampling methods with a minimal impact on wildlife has become paramount in complying with modern ethical standards and regulations, and to collect high-quality and unbiased data. We analysed the publication trends on non-invasive sampling in wildlife health and disease research and offer a comprehensive review on the different samples that can be collected non-invasively. We retrieved 272 articles spanning from 1998 to 2021, with a rapid increase in number from 2010. Thirty-nine percent of the papers were focussed on diseases, 58% on other health-related topics, and 3% on both. Stress and other physiological parameters were the most addressed research topics, followed by viruses, helminths, and bacterial infections. Terrestrial mammals accounted for 75% of all publications, and faeces were the most widely used sample. Our review of the sampling materials and collection methods highlights that, although the use of some types of samples for specific applications is now consolidated, others are perhaps still underutilised and new technologies may offer future opportunities for an even wider use of non-invasively collected samples. Full article

The objective of the study was to elucidate the stearoyl-coenzyme A desaturase (SCD1)-dependent gene network of c9, t11-CLA biosynthesis in MAC-T cells from an energy metabolism perspective. The cells were divided into the CAY group (firstly incubated with CAY10566, a chemical inhibitor of SCD1, then incubated with trans-11-octadecenoic acid, (TVA)), the TVA group (only TVA), and the control group (without CAY, TVA). The c9, t11-CLA, and TVA contents were determined by gas chromatography. The mRNA levels of SCD1 and candidate genes were analyzed via real-time PCR. Tandem mass tag (TMT)-based quantitative proteomics, bioinformatic analysis, parallel reaction monitoring (PRM), and small RNA interference were used to explore genes involved in the SCD1-dependent c9, t11-CLA biosynthesis. The results showed that the SCD1 deficiency led by CAY10566 blocked the biosynthesis of c9, t11-CLA. In total, 60 SCD1-related proteins mainly involved in energy metabolism pathways were primarily screened by TMT-based quantitative proteomics analysis. Moreover, 17 proteins were validated using PRM analysis. Then, 11 genes were verified to have negative relationships with SCD1 after the small RNA interference analysis. Based on the above results, we concluded that genes involved in energy metabolism pathways have an impact on the SCD1-dependent molecular mechanism of c9, t11-CLA biosynthesis. Full article

Welfare and management decisions for unowned free-ranging cats in urban environments should no longer be based on knowledge about behavioural ecology of solitary cats living and breeding in more natural ‘wild’ environments. We provide evidence that urban free-ranging domestic cats in the Anthropocene have responded to rapidly changing environments, such as abundance of food and higher population densities of conspecifics by adapting their behaviour (behavioural plasticity—the ability of a genotype (individual) to express different behaviours according to its environment) and social organisation to living in complex social groups, especially those living in colonies. Urban free-ranging cats are now more social, as demonstrated by different breeding patterns, lower infanticide, more frequent affiliative interactions in general, and different spatial groupings. We argue that this knowledge should be disseminated widely, and inform future research and strategies used to manage free-ranging cats across environments. Understanding behavioural plasticity and other recently evolved traits of domestic cats may lead to management strategies that maximise health and welfare of cats, wildlife, and humans—otherwise domestic cat behaviour may be ‘misunderstood’. Importantly, interdisciplinary research using expertise from biological and social sciences, and engaging human communities, should evaluate these management strategies to ensure they maintain optimal welfare of free-ranging domestic cats while preserving biodiversity and protecting wildcats. Full article

Increasing the levels of n-3 fatty acids (FA) in dairy products is an important goal in terms of enhancing the nutritional value of these foods for the consumer. The purpose of this research was to evaluate the effects of linseed and algae oil supplements in ovine isoenergetic diets on healthy milk fatty acid composition, mainly n-3. Seventy-two Churra dairy ewes were divided and randomly assigned to four experimental treatments for 6 weeks. The treatments consisted of a TMR (40:60 forage:concentrate ratio) that varied according to the inclusion of different types of fat (23 g/100 g TMR): hydrogenated palm oil (control), linseed oil (LO), calcium soap of linseed oil (CaS-LO) and marine algae oil (AO). The most effective lipid supplement to increase n-3 FA in milk was AO. 22:6 n-3 and total n-3 PUFA content increased from 0.02 and 0.60% (control) to 2.63 and 3.53% (AO), respectively. All diets supplemented with n-3 FA diminished the content of saturated FA in milk and its atherogenic index, while the levels of trans-11 18:1 and cis-9 trans-11 18:2 significantly increased. Overall, the enhancement of n-3 FA in ewe’s milk would be advantageous for the manufacture of nutritionally improved cheeses. Full article

Ninety crossbreed bulls (349.5 ± 8.25 kg initial weight) were used in an 87day trial to compare the effects of a blend of essential oils plus 25-hydroxy-Vit-D3 (EO + HyD) versus the combination of monensin with virginiamycin (MON + VM) on feedlot growth performance and carcass characteristics. Dietary treatments (nine replicates/treatment) were supplemented with 40 mg/kg diet dry matter of MON + VM (equal parts) or with 120.12 mg/kg diet dry matter of a combination of standardized mixture of essential oils (120 mg) plus 0.12 mg of 25-hydroxy-vitamin-D3 (EO + HyD). There were no treatment effects on dry matter intake (DMI, p = 0.63). However, the coefficient of variation in day-to-day DMI was greater for EO + HyD than for MON + VM (11.4% vs. 3.88%, p = 0.04). There were no treatment effects (p ≥ 0.17) on daily weight gain, gain-to-feed ratio, and estimated dietary net energy. Cattle supplemented with EO + HyD had greater Longissimus muscle area (7.9%, p < 0.01) and estimated retail yield (1.6%, p = 0.03), and tended to have heavier (1.7%, p = 0.10) carcass weight. Differences among treatments in dressing percentage, fat thickness, kidney–pelvic–heart fat, and marbling score were not appreciable (p > 0.10). It is concluded that growth performance response and dietary energetic are similar for finishing cattle supplemented with EO + HyD vs. MON + VM. However, compared with MON + VM, supplementation with EO + HyD during the finishing phase may improve carcass Longissimus area and carcass yield. Full article

The aim of the study was to determine the effects of genotype and sex on carcass composition and selected meat quality parameters of Dworka and Pekin ducks after two reproductive seasons. The research material consisted of 24 carcasses of Dworka ducks (breeding strain D11) and 24 carcasses of Pekin ducks from the herd of genetic resources (French Pekin, strain P-9). After cooling the carcasses (18 h, 2 °C), the pH values and electrical conductivity of the pectoral and leg muscles were determined, and then the carcass was dissected using a simplified method. After dissection, the pectoral and leg muscles were sampled for quality characteristics assessment. The Dworka ducks from breeding strain D11 compared to Pekin duck from conservative strain P9 were characterized by significantly (p < 0.05) higher carcass weight, protein and salt content, thermal drip, yellowness, thickness of perymisium and endomysium and lower water content of the pectoral muscle. Dworka ducks were also characterized by higher protein, salt and collagen content and higher electrical conductivity (EC₂₄) of leg muscles than Pekin ducks. Regardless of genotype, male ducks had a higher gutted carcass weight with neck, salt content, muscle fiber cross-sectional area, fiber perimeter and diameters of pectoral muscle, and also higher protein and collagen, and lower fat content, pH₂₄ and electrical conductivity of leg muscles. The genotype by sex interaction was significant (p < 0.05) for water, protein, fat content, perimisium thickness, cohesiveness, springiness of the pectoral major muscle, and for water content and protein of the leg muscles. The results obtained in this study demonstrate the effects of genotype and sex on the nutritional value and some technological parameters of duck meat. The studied ducks after two reproductive seasons satisfactorily meet the requirements of duck meat for consumers and duck meat processing plants. Full article

The sustainability of the Aquaculture industry relies on optimising diets to promote nitrogen retention and maximise fish growth. The aim of this study was to assess how different dietary formulations influence the bioavailability and metabolic fate of distinct amino acids in gilthead seabream juveniles. Amino acids (lysine, tryptophan, and methionine) were selected based on their ketogenic and/or glucogenic nature. Seabream were fed practical diets with different protein (44 and 40%) and lipid contents (21 and 18%): 44P21L, 44P18L, 40P21L, and 40P18L. After three weeks of feeding, the fish were tube-fed the correspondent diet labelled with ¹⁴C-lysine, ¹⁴C-tryptophan, or ¹⁴C-methionine. The amino acid utilisation was determined based on the evacuation, retention in gut, liver, and muscle, and the catabolism of the tracer. The metabolic fate of amino acids was mainly determined by their nature. Tryptophan was significantly more evacuated than lysine or methionine, indicating a lower availability for metabolic purposes. Methionine was more retained in muscle, indicating its higher availability. Lysine was mainly catabolised, suggesting that catabolism is preferentially ketogenic, even when this amino acid is deficient in diets. This study underpins the importance of optimising diets considering the amino acids’ bioavailability and metabolic fate to maximise protein retention in fish. Full article

Sexual dimorphism is common in most vertebrate species and has diverse manifestations. The study of sexual dimorphism has critical significance for evolutionary biological and ecological adaptation. In this study, we analysed the morphometric data of Hynobius maoershanensis, a rare and endangered species, to examine sexual dimorphism in size and shape. A total of 61 H. maoershanensis individuals (9 adult females and 52 adult males) were used in this study. We measured 14 morphological variables and weight of each individual. Analysis of covariance using snout–vent length (SVL) as the covariate showed significant differences in head width (HW), tail length (TL), tail height (TH), forelimb length (FLL), hindlimb length (HLL) and space between axilla and groin (AGS) between the male and female. The female AGS was greater than that of the male, whereas males had greater HW, TL, TH, FLL and HLL than females. The findings show that sexual dimorphism is present in terms of shape but not in terms of size. The wider head of the male could improve mating success, and its thicker limbs and longer tail might facilitate courtship. The females’ wider AGS may increase reproductive output. Our results support sexual dimorphism in H. maoershanensis, which could be explained by the sexual selection and fecundity theory hypothesis. Full article

With many advancements, technologies are now capable of recording non-human animals’ location, heart rate, and movement, often using a device that is physically attached to the monitored animals. However, to our knowledge, there is currently no technology that is able to do this unobtrusively and non-invasively. Here, we review the history of technology for use with animals, recent technological advancements, current limitations, and a brief introduction to our proposed novel software. Canadian tech mogul EAIGLE Inc. has developed an artificial intelligence (AI) software solution capable of determining where people and assets are within public places or attractions for operational intelligence, security, and health and safety applications. The solution also monitors individual temperatures to reduce the potential spread of COVID-19. This technology has been adapted for use at the Toronto Zoo, initiated with a focus on Sumatran orangutans (Pongo abelii) given the close physical similarity between orangutans and humans as great ape species. This technology will be capable of mass data collection, individual identification, pose estimation, behaviour monitoring and tracking orangutans’ locations, in real time on a 24/7 basis, benefitting both zookeepers and researchers looking to review this information. Full article

Prebiotics are known to have many beneficial effects on intestinal health by modulating the gut microbiota composition, thereby affecting epithelial cell proliferation and metabolism. This study had two aims: (1) to identify the protein constituents in the cecal mucosa of 50-day-old healthy (PIC × Penarlan P76) barrows, and (2) to assess the effects of 4% inclusion of dried chicory root in a cereal-based diet on the cecal mucosa proteome changes. Pigs (eight per group) were randomly allotted to the groups and were fed a control diet from the tenth day of life (C) or a diet supplemented with 4% of died chicory root (CR), for 40 days. At the age of 50 days, animals were sacrificed and cecal tissue samples were collected. It was found that feeding a CR diet significantly decreased the expression of 16 cecal mucosa proteins. Among them, fifteen proteins were down-regulated, while only one (KRT20) was shown to be up-regulated when compared to the C group. Dietary supplementation with CR caused down-expression of metabolism-associated proteins including enzymes involved in the process of glycolysis (G6PD, TPI1, ALDH9A1, CKMT1 and AKR1A1) as well as those engaged in transcriptional and translational activity (PRPF19, EEF1G) and several structural proteins (ACTR3, KRT77, CAP1 and actin). From our findings, it is possible to conclude that dietary chicory root at 4% had beneficial effects on the gut health of pigs as indicated by a changed abundance of certain cecal proteins such as KRT20, SERPINB1, HSP27, ANAXA2 and ANAXA4. Full article

The ocellated skink (Chalcides ocellatus) is a widespread lizard, naturally distributed between the Maghreb and coastal Pakistan, with few insular populations in the Mediterranean coastal area. Some populations of this species have also been recorded in peninsular Italy, Campania and Southern Tuscany due to accidental introductions via touristic and commercial routes. In this work, we conducted genetic analyses on mitochondrial DNA COXI, cytb and 16S mtDNA genes on a sample of Italian insular and peninsular populations. Differently from what previously suggested, the nucleus in Portici (Southern Italy) may have originated from Sardinia. The intense trade and touristic traffic between Sardinia and Southern Tuscany may have been responsible for the introduction of this lizard also to Central Italy. Full article

Ticks transmit pathogens to animals and humans more often than any other arthropod vector. The rural economy of Pakistan mainly depends on livestock farming, and tick infestations cause severe problems in this sector. The present study aimed to molecularly characterize the Anaplasma spp. in hard ticks collected from six districts of Khyber Pakhtunkhwa, Pakistan. Ticks were collected from various livestock hosts, including cattle breeds (Holstein-Friesian, Jersey, Sahiwal, and Achai), Asian water buffaloes, sheep, and goats from March 2018 to February 2019. Collected ticks were morphologically identified and subjected to molecular screening of Anaplasma spp. by amplifying 16S rDNA sequences. Six hundred seventy-six ticks were collected from infested hosts (224/350, 64%). Among the nine morphologically identified tick species, the highest occurrence was noted for Rhipicephalus microplus (254, 37.6%), followed by Hyalomma anatolicum (136, 20.1%), Rhipicephalus haemaphysaloides (119, 17.6%), Rhipicephalus turanicus (116, 17.1%), Haemaphysalis montgomeryi (14, 2.1%), Hyalomma dromedarii (11, 1.6%), Haemaphysalis bispinosa (10, 1.5%), Hyalomma scupense (8, 1.2%), and Haemaphysalis kashmirensis (8, 1.2%). The occurrence of tick females was highest (260, 38.5%), followed by nymphs (246, 36.4%) and males (170, 25.1%). Overall, the highest occurrence of ticks was recorded in the Peshawar district (239, 35.3%), followed by Mardan (183, 27.1%), Charsadda (110, 16.3%), Swat (52, 7.7%), Shangla (48, 7.1%), and Chitral (44, 6.5%). Among these ticks, Anaplasma marginale was detected in R. microplus, R. turanicus, and R. haemaphysaloides. The 16S rDNA sequences showed high identity (98–100%) with A. marginale reported from Australia, China, Japan, Pakistan, Thailand, Uganda, and the USA. In phylogenetic analysis, the sequence of A. marginale clustered with the same species reported from Australia, China, Pakistan, Thailand, Uruguay, and the USA. Further molecular work regarding the diversity of tick species and associated pathogens is essential across the country. Full article

Ewes often lamb over extended periods so the level of nutrition during pregnancy and lambing may be suboptimal for ewes that conceived later during mating. Predicting lambing dates would allow cohorts of ewes with similar gestational ages to be managed more precisely to achieve targets for ewe nutrition, feed on offer, mob sizes and access to shelter to improve lamb survival. The interactions between ewes and rams during mating have been used to predict the time of oestrus and lambing dates successfully, but this has not been tested at a commercial scale. In this study, proximity sensors were used to measure interactions between inexperienced Merino ewes (n = 317) and experienced rams (n = 9) during a 27-day mating period under commercial production conditions. When the gestation length was assumed to be 150 days, 91% of lambing dates were predicted within ±6 days of the actual birth date of lambs and 84% of lambing dates were predicted within ±3 days. The use of proximity sensors during mating was an effective means of predicting lambing dates, and there was no significant difference in accuracy for single bearing verses multiple bearing ewes. However, DNA parentage data showed the ram corresponding with the maximum daily interactions ratio was the sire for only 16% of all progeny, suggesting they could not be used to indicate the sire of the progeny. Full article

The acquisition of mammalian sperm motility is a main indicator of epididymal sperm maturation and helps ensure fertilization. Poor sperm motility will prevent sperm cells from reaching the fertilization site, resulting in fertilization failure. To investigate the proteomic profiling of normal and poorly motile buffalo spermatozoa, a strategy applying liquid chromatography tandem mass spectrometry combined with tandem mass targeting was used. As a result, 145 differentially expressed proteins (DEPs) were identified in poorly motile spermatozoa (fold change > 1.5), including 52 upregulated and 93 downregulated proteins. The upregulated DEPs were mainly involved in morphogenesis and regulation of cell differentiation. The downregulated DEPs were involved with transport, oxidation-reduction, sperm motility, regulation of cAMP metabolism and regulation of DNA methylation. The mRNA and protein levels of PRM1 and AKAP3 were lower in poorly motile spermatozoa, while the expressions of SDC2, TEKT3 and IDH1 were not correlated with motility, indicating that their protein changes were affected by transcription or translation. Such changes in the expression of these proteins suggest that the formation of poorly motile buffalo spermatozoa reflects a low efficiency of energy metabolism, decreases in sperm protamine proteins, deficiencies in motility-related proteins, and variations in tail structural proteins. Such proteins could be biomarkers of poorly motile spermatozoa. These results illustrate some of the molecular mechanisms associated with poorly motile spermatozoa and provide clues for finding molecular markers of these pathways. Full article