Animals, Volume 13, Issue 17 (September-1 2023) – 125 articles

A new pterygosomatid mite species, Neopterygosoma robertmertensi sp. n. (Acariformes: Pterygosomatidae) was collected from two specimens of Liolaemus robertmertensi (Liolaemidae) from Argentina. This new species is described based on active stages: adults (female and male) and juveniles (deutonymphs, protonymphs, and larvae) and quiescent stages (nymphchrysalis, deutochrysalis and imagochrysalis). The changes in morphological characters that occur during the ontogeny of N. robertmertensi have been analyzed in detail. A difference in larval sex morphology was observed for the first time in the family Pterygosomatidae (female larvae differ from male larvae in terms of the shape and size of the idiosoma and the position of the genital area). This new mite species is most similar to N. cyanogasteri but can be distinguished by the presence of different leg chaetotaxy patterns of genua IV and femora IV, four to six genital setae, three to five dorsomedial setae, and two to three ventromedial setae. Phylogenetic analysis was conducted based on 120 morphological characters of all Neopterygosoma spp. and four outgroup species using the maximum parsimony approach. The results indicated that this species is nested within mites of the chilensis group of Neopterygosoma associated with host species of the section chiliensis of Liolaemus s. str. An updated diagnosis of the chilensis group of Neopterygosoma and an identification key for all species of this genus has been provided. Full article

This study investigates the chemical composition of soyhulls (SHs) as an alternative feed ingredient and their effect on nutrient and amino acid (AA) digestibility in laying hens during peak production. A total of 200 golden brown hens (28 weeks old) were subjected to random allocation across 5 dietary treatments: a corn–soybean meal (SBM) reference diet and 4 test diets with 25% SHs from different mills (SH1, SH2, SH3, and SH4). Each treatment was replicated four times with ten birds per replicate. Digesta samples were collected during three phases (28–32, 32–36, and 36–40 weeks of age) to measure apparent metabolizable energy (AME), the apparent ileal digestibility (AID) of nutrients, and the standard ileal digestibility (SID) of AAs. The SBM diet had 30.0% crude protein (CP) and 3.78% crude fiber (CF), while the SH diets had 21.0 to 21.5% CP and 11.6% CF. The findings revealed that the AME was lower (p < 0.05) with SH diets (2404 kcal/kg) compared to the SBM diet (2627 kcal/kg) in all three phases. The SH diets had a lower AID of dry matter (DM), crude protein (CP), ash, ether extract (EE), and crude fiber (CF) than the SBM diet by an average of 2.88, 2.25, 4.93, 4.99, and 3.36%, respectively. The AID of nitrogen-free extract (NFE) was higher in the SH diets than the SBM diet by 3.42% in all three phases (p < 0.05). The SH diets had lower uric acid excretion (about 66.93 mg/100 mL) than the SBM diet (about 76.43 mg/100 mL) on average in all three phases. The SH diets had a lower SID of arginine, histidine, isoleucine, lysine, cysteine, valine, and tyrosine than the SBM diet by 2 to 10%, while the SID of methionine was higher in the SH diets than the SBM diet by 2.2% on average in all three phases (p < 0.05). The SH from Sadiq Brother Feed (SH1) had the highest AME and AID of DM, ash, CP, EE, CF, and the SID of AA among the SH diets. These results indicate that SH can partially replace SBM in laying hen diets, but the source and quality of SH should be considered. Full article

Stool sampling is a useful tool for diagnosing gastrointestinal disease in veterinary medicine. The sub-clinical disease burden of Salmonella spp. in cattle can become significant for farmers. However, current methods of faecal sampling in a rural setting for diagnosis are not consistently sufficient for the preservation of Salmonella spp. in faeces. This study evaluated the use of a commercial stool storage kit for bacterial preservation in cow faecal samples compared to unpreserved stools placed into refrigeration at different time-points. A stool sample was collected per-rectum from one apparently healthy Holstein–Freisen cow. The sample was weighed and aliquoted into two sterile Falcon tubes and into two commercial kit tubes. The aliquots were then placed into refrigeration at 4 °C at 0, 24, and 96 h after processing. One commercial kit tube was not aliquoted and remained at ambient temperature. After 2 weeks, DNA was extracted from the samples and analysed using endpoint PCR, revealing a sub-clinical infection with Salmonella spp. The bacterium was best preserved when the stool was stored in the commercial kit at ambient temperature and re-homogenised immediately prior to DNA extraction. The unpreserved stool did not maintain obvious levels of Salmonella spp. after 24 h at ambient temperature. This commercial kit should be considered for use in the diagnosis of salmonellosis in cattle. Full article

The Cashmere goat is an excellent local goat breed in Gansu Province of China, and it is expected to improve cashmere production and cashmere quality through selection and breeding to enhance its commercial value. Keratin-associated proteins (KAPs) play an important role in maintaining wool structure. The gene encoding the keratin-associated protein 22-2 (KAP22-2) gene has been identified in selected species other than goats, such as humans, mice, and sheep. In this study, the sequence of the sheep KAP22-2 gene (KRTAP22-2) was aligned into the goat genome, and the sequence with the highest homology was assumed to be the goat KRTAP22-2 sequence and used to design primers to amplify the goat gene sequence. A total of 356 Longdong Cashmere goats (Gansu Province, China) were used for screening of genetic variants. Four specific bands were detected by polymerase chain reaction-single-stranded conformational polymorphism (PCR-SSCP) analysis, and they formed a total of six band types individually or in combination. Four alleles were identified by DNA sequencing of PCR amplification products. A total of four single nucleotide polymorphic sites (SNPs) were detected in the four sequenced KRTAP22-2 alleles. Two of them are in the 5’UTR region and the other two are in the coding region, and the variants in the coding region are all non-synonymous mutations. In addition, there was a 6 bp length variation in allele C. The gene was expressed in the cortical layer of primary and secondary hair follicles, the inner root sheath, as well as hair papillae and hair maternal cells in goats. The results of the correlation analysis between genotypes and cashmere traits showed that after excluding genotypes with a gene frequency of less than 5%, the mean fiber diameter (MFD) of cashmere was significantly higher in the AB genotype than in the AA and AC genotypes. That is, the KRTAP22-2 gene variants are associated with mean fiber diameter in cashmere. The above results suggest that the goat KRTAP22-2 variant can be utilized as a molecular marker candidate gene for cashmere traits. Full article

The CSN1S2 gene encodes αs2-casein, the third most abundant protein in camel milk. Despite its importance in foals, human nutrition, and dairy processing, the CSN1S2 gene in camels has received little attention. This study presents the first complete characterization of the CSN1S2 gene sequence in Old-World camels (Camelus bactrianus and Camelus dromedarius). Additionally, the gene promoter, consisting of 752 bp upstream of exon 1, was analyzed. The entire gene comprises 17 exons, ranging in length from 24 bp (exons 4, 8, 11, and 13) to 280 bp (exon 17). Interesting was the identification of the exon 12 in both species. The promoter analysis revealed 24 putative binding sites in the Bactrian camel and 22 in dromedary camel. Most of these sites were typical elements associated with milk protein, such as C/EBP-α, C/EBP-β, Oct-1, and AP1. The SNP discovery showed relatively high genetic diversity compared to other camel casein genes (CSN1S1, CSN2, and CSN3), with a total of 34 polymorphic sites across the two species. Particularly noteworthy is the transition g.311G>A in the CSN1S2 promoter, creating a new putative consensus binding site for a C/EBP-β in the Bactrian camel. At the exon level, two novel variants were found. One was detected in exon 6 of the Bactrian camel (g.3639C>G), resulting in an amino acid replacement, p.36Ile>Met. The second variant was found in noncoding exon 17 of dromedary CSN1S2 (g.1511G>T). Although this mutation occurs in the 3′-UnTranslated Region, it represents the first example of exonic polymorphism in the CSN1S2 for this species. This SNP also affects the binding sites of different microRNAs, including the seed sequence of the miRNA 4662a-3p, highlighting its role as a regulatory factor for CSN1S2 gene. A PCR-RFLP was set up for genotyping a dromedary Tunisian population (n = 157), and the minor allele frequency was found to be 0.27 for the G allele, indicating a potential yield improvement margin. The interspersed elements (INEs) analysis revealed 10 INEs covering 7.34% and 8.14% of the CSN1S2 sequence in the Bactrian and dromedary camels, respectively. Furthermore, six elements (A, B, F, H, I, and L) are shared among cattle and camels and are partially found in other ruminants, suggesting a common ancestral origin of these retrotransposons. Conversely, elements C, D, E, and G are specific to camels. Full article

Chronic inflammatory enteropathies (CIEs) in dogs involve the infiltration of gastrointestinal tissue with inflammatory cells. This study aimed to assess the sensitivity of serum and fecal 3-bromotyrosine (3-BrY) concentrations in dogs with CIE. The difference in 3-BrY concentrations in dogs with different gastrointestinal (GI) pathological changes was also assessed. In total, 68 dogs with CIE were enrolled in the study. The median serum 3-BrY concentration was 3.3 µmol/L, while the median 3-day mean and maximum fecal 3-BrY concentrations were 38.9 and 63.2 mmol/g of feces, respectively. The median serum C-reactive protein concentration was 45.0 mg/L. The median 3-day mean and maximum fecal α₁-proteinase inhibitor concentrations were 6.1 and 9 µg/g of feces, respectively. Increased 3-BrY concentrations were observed in 90.9% of CIE dogs based on serum concentrations, 75.8% based on mean fecal concentrations, and 69.4% based on maximum fecal concentrations. A weak correlation (ρ = 0.31, p < 0.0118) was found between serum CRP and serum 3-BrY concentrations. There was no correlation between the canine chronic enteropathy clinical activity index and serum or fecal 3-BrY concentrations (p > 0.05). Additionally, no significant difference in serum or fecal 3-BrY concentrations was found among CIE dogs with different GI pathological changes (p > 0.05). In conclusion, dogs with CIE have increased 3-BrY concentrations in serum and fecal samples. However, 3-BrY concentrations may not accurately indicate the severity of gastrointestinal inflammation. Full article

Fixed-timed artificial insemination (FTAI) protocols for beef cattle in South America are primarily based on estradiol esters and intravaginal progesterone-releasing devices (IVPD). The objective of this study was to determine the optimal gonadotropin-releasing hormone (GnRH)-based protocol as an alternative to the use of estrogen-based protocols in grazing Bos taurus suckling beef cows. All cows received an IVPD on the day of protocol initiation and prostaglandin F_2α (PG) plus equine chorionic gonadotropin (eCG) treatments at the time of IVPD removal. In Experiment 1, cows (n = 235) were randomly assigned to one of four treatments: (i) 7-day estradiol = 2 mg of estradiol benzoate (EB) at IVPD insertion on Day 9 and 1 mg of estradiol cypionate (ECP) at IVPD removal on Day 2; (ii) 7-day GnRH = 10 µg of GnRH at IVPD insertion on Day 10, IVPD removal on Day 3 and GnRH at FTAI; (iii) 7 & 7 estradiol = PG at IVPD insertion on Day 16, EB on Day 9 and ECP at IVPD removal on Day 2; (iv) 7 & 7 GnRH = PG at IVPD insertion on Day 17, GnRH on Day 10, IVPD removal on Day 3 and GnRH at FTAI. In Experiment 2, cows (n = 462) were randomly assigned to one of four treatments: (i) 6-day estradiol = EB at IVPD insertion on Day 9, IVPD removal on Day 3 and GnRH at FTAI; (ii) 7-day estradiol; (iii) 7-day GnRH; (iv) 7 & 7 GnRH. In Experiment 1, plasma progesterone concentrations and percentage of cows with a corpus luteum (CL) at IVPD removal, and pregnancy per AI (P/AI) were greater for cows subjected to GnRH-based protocols compared with cows subjected to estrogen-based protocols (p < 0.01). In Experiment 2, cows subjected to the 7 & 7 GnRH protocol had the greatest P/AI (p < 0.01). In summary, GnRH-based FTAI protocols resulted in similar or greater P/AI compared to estrogen-based FTAI protocols in grazing postpartum Bos taurus suckled beef cows. The greatest P/AI was attained with the 7 & 7 GnRH protocol. Full article

The genus Alytes consists of six primitive terrestrial-breeding species (five of them endangered or present in restricted distributions) with unique male parental care. Alytes obstetricans was used as a model for the development of assisted reproductive technologies for gamete collection, artificial fertilization and captive-rearing of embryos. Sperm collection was achieved with human chorionic gonadotropin (hCG), but only 17 to 50% of males responded to the hormone. In addition, an effect of captivity on sperm quality was observed. A combination of gonadotropin-releasing hormone agonist (GnRHa) and hCG was tested for induction of ovulation, and 100% of females responded to the treatment. Recently collected sperm was used in artificial fertilization, and 10% of eggs showed complete embryo development. To design a method for captive-rearing of embryos, natural clutches (n = 13) were removed from males and divided into two sub-clutches. One was kept with all eggs together and in the other, the egg string was cut and eggs were split individually. All sub-clutches with eggs kept separated presented embryo survival (ranging from 27 to 61%) that hatched normally, while only one sub-clutch with embryos maintained together had some embryo survival (36%). These results may contribute to the management of this and related species. Full article

Birds wintering in the northern Palearctic compensate for substantial energy losses and prepare for a food deficit in winter by adjusting their foraging behavior. Apart from weather conditions, interspecific competition also drives hunting strategies. To describe this phenomenon, we observed the behavior of two sympatrically wintering raptor species: the Common Buzzard and the Rough-legged Buzzard. The study was carried out in east-central Poland during four seasons on a study plot where the densities of both species were high. Interspecific differences were detected in the use of available hunting sites. Rough-legged Buzzards conspicuously avoided using fence posts for scanning the surroundings and spent the most time standing on the ground. Common Buzzards more often used trees for this purpose when the snow cover was thick. Thicker snow cover resulted in fewer attempted attacks on prey in both species and caused Common Buzzards to change their hunting sites less frequently. The study also showed that the more often a bird changed its hunting site, the greater the number of attempted attacks. The outcome is that the ultimate effectiveness of hunting is mediated by the overview of the foraging area from different heights and perspectives, not by the type of hunting site. Snow cover was the most important factor in modifying foraging behavior and possibly intensifying interspecific competition. Full article

The subgenus Ochthomantis is an obligate forest and stream-dwelling group of mantellid frogs, endemic to Madagascar, with six species currently recognized. However, this group suffers from ongoing taxonomic confusion due to low numbers of examined specimens, and failure to consider morphological variation from development and sexual dimorphism. Here, we examined the morphology of 637 sexed adult specimens collected by us in the field and from other museum collections. We also sequenced a DNA fragment of the 16S mtDNA gene for each lineage to determine congruence between morphological and molecular data sets and to help delimit species. Our results demonstrate that the subgenus Ochthomantis includes eleven valid species: five already recognized, M. catalai and M. poissoni that we resurrect from synonymy, and four new species which we describe for the first time here. In some analyses, Mantidactylus majori groups with other Mantidactylus subgenera, so we do not consider it a member of the subgenus Ochthomantis in this study. All species have restricted distributions and elevational ranges in the humid forests of Madagascar. This study demonstrates the utility of assessing cryptic species using both diagnostic morphological characters and molecular data. The discovery of this new cryptic biodiversity, and the taxonomic revision herein, will likely require conservation activities for those species with the most restricted distributions. Full article

This paper provides a comprehensive discussion on the physiological impacts of hypoxic training, its benefits to endurance performance, and a rationale for utilizing it to improve performance in the equine athlete. All exercise-induced training adaptations are governed by genetics. Exercise prescriptions can be tailored to elicit the desired physiological adaptations. Although the application of hypoxic stimuli on its own is not ideal to promote favorable molecular responses, exercise training under hypoxic conditions provides an optimal environment for maximizing physiological adaptations to enhance endurance performance. The combination of exercise training and hypoxia increases the activity of the hypoxia-inducible factor (HIF) pathway compared to training under normoxic conditions. Hypoxia-inducible factor-1 alpha (HIF-1α) is known as a master regulator of the expression of genes since over 100 genes are responsive to HIF-1α. For instance, HIF-1-inducible genes include those critical to erythropoiesis, angiogenesis, glucose metabolism, mitochondrial biogenesis, and glucose transport, all of which are intergral in physiological adaptations for endurance performance. Further, hypoxic training could conceivably have a role in equine rehabilitation when high-impact training is contraindicated but a quality training stimulus is desired. This is achievable through purpose-built equine motorized treadmills inside commercial hypoxic chambers. Full article

Avian malaria and leucocytozoonosis can cause fatal diseases, whereas avian trypanosomiasis is reported to be harmless in chickens. Backyard chickens can be infected by several pathogens, including blood parasites, that may shed to industrial poultry production, with a consequently higher economic impact. This study aimed to investigate the presence of several blood parasites (Plasmodium, Leucocytozoon and Trypanosoma) in backyard chickens raised in Southern Thailand, using PCR-based detection and microscopic methods. From June 2021 to June 2022, 57 backyard chickens were sampled. Fresh thin blood smears were prepared from 11 individuals, and buffy coat smears were prepared from 55 of them. Both thin blood smears and buffy coat smears were used for microscopic analysis. Two nested PCR protocols that amplify a fragment of cytochrome b (cytb) and small subunit rRNA (SSU rRNA) genes were used to identify Haemosporida and Trypanosoma parasites, respectively. The number of positive samples was higher with the application of nested PCR than when buffy coat smears were used. Three new Plasmodium lineages (GALLUS47-49) and thirteen Leucocytozoon lineages (GALLUS50-62) were found. Trophozoites, meronts and gametocytes of Plasmodium gallinaceum (GALLUS01) were present in one thin blood smear. All thin blood smears revealed Leucocytozoon infections, but only three samples were a single infection. These three samples revealed the presence of fusiform host cell–parasite complexes, of which the morphological features resembled those of Leucocytozoon macleani (possible synonym is Leucocytozoon sabrazesi), while the cytb showed that this parasite is closely related to the lineage GALLUS06-07, described as Leucocytozoon schouteni. The Trypanosoma prevalence was 33.33%; it was present in only one of the thin blood smears, and it resembles Trypanosoma calmettei. This study showed the prevalence of a high diversity of Plasmodium (64.91%) and Leucocytozoon (89.47%) in Thai chickens. Both nested-PCR and buffy coat smear can be used as the diagnostic tool for the testing of Plasmodium, Leucocytozoon and Trypanosoma for parasitic control in backyard chickens and poultry farms. The information on the parasite species that can be found in chickens raised in Southern Thailand was also considered as the baseline information for further study. Full article

Sport performances judged subjectively often suffer from systematic errors due to biases, with the sport of equestrian dressage being no exception. This study examines whether international dressage judges display systematic errors while evaluating elite horse-rider combinations. Data from seven 5* Grand Prix dressage events between May 2022 and April 2023 were analyzed (510 judges’ scores) using Multivariable Linear Regression Analysis. Five predictor variables—Home, Same Nationality, Compatriot, FEI Ranking and Starting Order—were studied in relation to Total Dressage Score (TS). The model accounted for 44.1% of TS variance; FEI Ranking, Starting Order, Compatriot, Same Nationality, and Home were statistically significant (p < 0.001). Judges exhibited nationalistic and patriotism-by-proxy biases, awarding significantly higher scores to riders from their countries (p < 0.001). FEI Ranking and Starting Order also influenced scores significantly (p < 0.001). These biases, combined, created a cascade effect benefiting a specific group of riders. To address this, measures should be taken to develop a more objective judging system that is based on unequivocal, transparent and evidence-based criteria and supports the continuous development of a fair, sustainable, equine welfare orientated sport that fosters societal acceptance, Full article

Support and management of second-intention wound healing involves frequent dressing changes having different properties. Dressings can range from simple ones, such as nonadherent dressings, to more complex options, such as foam, hydrocolloid, alginate or negative pressure dressings. Seven cats were enrolled in the study with a total of nine wounds of various sizes with different etiology sizes and localizations. Three methods of local ozone administration were used to cover more of the ozone properties in the treatment of wounds: bagging, perilesional subcutaneous infiltrations and lavages with ozonated saline. Evaluation of the healing process was performed by clinical observation and wound area measurements every seven days until the complete recovery of the patients. The results of this study should encourage clinicians to consider medical ozone as a new therapeutic approach with regenerative properties in the second-intention healing of cats presenting cutaneous wounds. Full article

Shiga toxin-producing E. coli (STEC) infections associated with wildlife are increasing globally, highlighting many ‘spillover’ species as important reservoirs for these zoonotic pathogens. A human outbreak of STEC serogroup O157 in 2015 in Scotland, associated with the consumption of venison meat products, highlighted several knowledge gaps, including the prevalence of STEC O157 in Scottish wild deer and the potential risk to humans from wild deer isolates. In this study, we undertook a nationwide survey of wild deer in Scotland and determined that the prevalence of STEC O157 in wild deer is low 0.28% (95% confidence interval = 0.06–0.80). Despite the low prevalence of STEC O157 in Scottish wild deer, identified isolates were present in deer faeces at high levels (>10⁴ colony forming units/g faeces) and had high human pathogenic potential based on whole genome sequencing and virulence gene profiling. A retrospective epidemiological investigation also identified one wild deer isolate from this study as a possible source of a Scottish human outbreak in 2017. These results emphasise the importance of food hygiene practices during the processing of wild deer carcasses for human consumption. Full article

This study evaluated the effect of a natural source of vitamin D₃ [1,25-(OH)₂D₃] and capsaicin (CAP) in the dietary supplementation of sows in the final phase (85–114 days) of gestation (Gest) and lactation (Lact) on the reproductive performance of the sows and health of piglets through two experiments (Exp I and II). In Exp I, 120 sows were subjected to four treatments: T1—control (without [1,25-(OH)₂D₃] and supplemental CAP); T2—3.5 µg 1,25-(OH)₂D₃/Gest/day and 7.0 µg Vit 1,25-(OH)₂D₃/Lact/day; T3—7.0 µg CAP/Gest/day and 14.0 µg CAP/Lact/day; T4—1.75 µg Vit 1,25-(OH)₂D₃ + 3.5 µg CAP/Gest/day and 3.5 µg 1,25-(OH)₂D₃ + 7.0 µg CAP/Lact/day. In Exp II, 200 sows were randomly blocked, factorial 2 × 2 (without or with Vit 1,25-(OH)₂D₃ and without or with CAP): T1—control (without Vit 1,25-(OH)₂D₃ and CAP); T2—3.5 µg Vit 1,25-(OH)₂D₃/Gest/day and 7.0 µg Vit 1,25-(OH)₂D₃/Lact/day; T3—7.0 µg CAP/Gest/day and 14.0 µg CAP/Lact/day; T4—3.5 µg Vit 1,25-(OH)₂D₃ + 7 µg CAP/Gest/day; and 7.0 µg Vit 1,25-(OH)₂D₃ + 14.0 µg CAP/Lact/day. The duration of delivery (3:48 vs. 4:57 h) and the percentage of stillbirths (5.37% vs. 7.61%) were improved (p < 0.05) in the group that received Vit 1,25-(OH)₂D₃ (Exp II) compared to the control group. Moreover, the dystocia rate decreased (p < 0.05) in Exp II, which received Vit 1,25-(OH)₂D₃ (4.21 vs. 27.63%), and in Exp I, which received the combination of Vit 1,25-(OH)₂D₃ + CAP (12 vs. 40%) compared to the respective control groups. Colostrum production was greater (p < 0.05) in sows that received Vit 1,25-(OH)₂D₃ supplementation compared to the control group, consequently resulting in higher colostrum intake (p < 0.05) of the piglets (330 vs. 258 g/piglet). The additives reduced the incidence of diarrhea (p < 0.05) in piglets (Exp I and II). Thus, the use of additives improved the reproductive performance of sows and contributed to litter growth. Full article

Reliable indicators of health status (heart rate, rectal temperature, blood marker, etc.) are of cornerstone importance in the daily practice of veterinary medicine. The reliability of a measurement assesses the variability that is associated with the variable to be measured itself vs. other sources of variation (measurement device, person performing the measurement, etc.). Quantitative and continuous indicators are numerous in practice and the determination of their reliability is a complex issue. In the absence of a gold standard approach, several indicators of reliability have been described and can be used depending on several assumptions, study design, and type of measurement. The aim of this manuscript is, therefore, to determine the applicability of commonly described reliability indicators. After a description of the different sources of errors of a measurement, a review of the different indicators that are commonly used in the veterinary field as well as their applicability, limitations, and interpretations is performed. Full article

Sex bias in biomedical and natural science research has been prevalent for decades. In many cases, the female estrous cycle was thought to be too complex an issue to model for, and it was thought to be simpler to only use males in studies. At times, particularly when studying efficacy and safety of new therapeutics, this sex bias has resulted in over- and under-medication with associated deleterious side effects in women. Many sex differences have been recognized that are unrelated to hormonal variation occurring during the estrous cycle. Sex bias also creates animal welfare challenges related to animal over-production and wastage, insufficient consideration of welfare (and scientific) impact related to differential housing of male vs female animals within research facilities, and a lack of understanding regarding differential requirements for pain recognition and alleviation in male versus female animals. Although many funding and government agencies require both sexes to be studied in biomedical research, many disparities remain in practice. This requires further enforcement of expectations by the Institutional Animal Care and Use Committee when reviewing protocols, research groups when writing grants, planning studies, and conducting research, and scientific journals and reviewers to ensure that sex bias policies are enforced. Full article

This review focused on the impact of F18⁺ E. coli on pig production and explored nutritional interventions to mitigate its deleterious effects. F18⁺ E. coli is a primary cause of PWD in nursery pigs, resulting in substantial economic losses through diminished feed efficiency, morbidity, and mortality. In summary, the F18⁺ E. coli induces intestinal inflammation with elevated IL6 (60%), IL8 (43%), and TNF-α (28%), disrupting the microbiota and resulting in 14% villus height reduction. Besides the mortality, the compromised intestinal health results in a 20% G:F decrease and a 10% ADFI reduction, ultimately culminating in a 28% ADG decrease. Among nutritional interventions to counter F18⁺ E. coli impacts, zinc glycinate lowered TNF-α (26%) and protein carbonyl (45%) in jejunal mucosa, resulting in a 39% ADG increase. Lactic acid bacteria reduced TNF-α (36%), increasing 51% ADG, whereas Bacillus spp. reduced IL6 (27%), increasing BW (12%). Lactobacillus postbiotic increased BW (14%) and the diversity of beneficial bacteria. Phytobiotics reduced TNF-α (23%) and IL6 (21%), enhancing feed efficiency (37%). Additional interventions, including low crude protein formulation, antibacterial minerals, prebiotics, and organic acids, can be effectively used to combat F18⁺ E. coli infection. These findings collectively underscore a range of effective strategies for managing the challenges posed by F18⁺ E. coli in pig production. Full article

Local breeds are the main reservoir of biodiversity of farm animals. According to FAO, they account for 87% of all described breeds in the world. To ensure that they are adequately protected, they should be continually monitored for genetic variation. Another crucial factor is reproduction, which is the most important guarantee of population growth. In the present study, genetic variation in 372 Polish White-Backed cows was determined using DNA microsatellite sequences, taking into account their parentage. Reproductive parameters were analysed as well, based on data from 3658 lactations of 1128 Polish White-Backed cows. The results indicate that despite the small initial population and the implementation of a moderate selection of animals, the existing population of Polish White-Backed cattle has a high level of genetic variation, reflected in the degree of heterozygosity (0.761). Regarding reproductive traits, despite their late age at first calving, Polish White-Backed cows were shown to be distinguished by very good fertility parameters in comparison to other breeds raised in Poland. These findings not only confirm the value of protecting local cattle breeds around the world but may also be of importance in developing selection indices for highly productive breeds, in which reproductive functioning should be one of the most important factors considered. Full article

The present study evaluated the effects of partially substituting fish meal (FM) with poultry by-product meal (PBPM) on the growth, muscle composition, and tissue biochemical parameters of coho salmon (Oncorhynchus kisutch) post-smolts. Five isonitrogenous (7.45% nitrogen) and isoenergetic (18.61 MJ/kg gross energy) experimental diets were made by substituting 0%, 10%, 20%, 40%, and 60% FM protein with PBPM protein, which were designated accordingly as PBPM0 (the control), PBPM10, PBPM20, PBPM40, and PBPM60, respectively. Each diet was fed to triplicates of ten post-smolts (initial individual body weight, 180.13 ± 1.32 g) in three floating cages three times daily (6:50, 11:50, and 16:50) to apparent satiation for 84 days. Both specific growth rate (SGR) and protein efficiency ratio did not differ significantly (p > 0.05) among the control, PBPM10, and PBPM20 groups, which were remarkably (p < 0.05) higher than those of the PBPM40 and PBPM60 groups. Feed conversion ratio varied inversely with SGR. The PBPM replacement had no remarkable effects on the morphological indices and proximal muscle components. The control and PBPM10 groups led to significantly higher muscle contents of leucine, lysine, and methionine than groups of higher PBPM inclusion. The groups of PBPM40 and PBPM60 obtained significantly (p < 0.05) higher serum alanine aminotransferase and aspartate aminotransferase activities than the control and low PBPM inclusion groups. The control group had significantly higher albumin and total cholesterol contents than the groups with PBPM inclusion. The control group had significantly higher triglycerides content than the PBPM60 group. The PBPM60 group had significantly lower contents of high-density lipoprotein, low-density lipoprotein, and total protein than the control and PBPM10 groups. The high PBPM replacement level up to 40% and 60% had adverse effects on hepatic malondialdehyde levels. The catalase and superoxide dismutase activities were not affected by low PBPM inclusion, but significantly decreased in high-PBPM-inclusion groups. Based on broken-line regression analysis of SGR and PER, the optimum dietary PBPM replacing level was evaluated to be 16.63–17.50% of FM protein for coho salmon post-smolts. Full article

Freya the Walrus, who often climbed onto docked boats to sunbathe and frolic, was euthanized by the Norwegian Department of Fisheries in the Oslo fjord in August 2022, sparking international outrage and media attention. Since walruses are social animals, and since the Anthropocene era of climate change has displaced animals from their Arctic homes, forcing them to migrate, we can expect more human–animal interactions at such places as marinas, where Freya met her end. This paper asks and attempts to answer how we can make such interactions just going forward? In cases such as Freya’s, we need to reconcile three competing interests: the animal’s interest in living a flourishing life as best they can in a changing climate; the public’s interest in a safe and fulfilling wildlife encounter with an animal they have come to know intimately enough to name and follow devotedly on social media; and interests in maintaining private property. Examining these interests through the philosophical lenses of co-sovereignty, capability, and individuality, however, will yield more just results for animals in similar situations of conflict and co-existence with humans in urban spaces. We argue that, going forward, state resources should be expended to safeguard the public from marina access if safety is a genuine concern, while private money should be spent by marinas to enact safe animal removal with a no-kill policy. Full article

The primary objective of this study was to investigate the association between the bulk tank somatic cell count (SCC) and dairy farmers’ knowledge about milk quality, mastitis control, and their socioeconomic characteristics. Additionally, we estimated the association between the bulk tank SCC and bulk tank milk hygienic quality in dairy herds. Bulk tank milk samples from 120 dairy herds enrolled in the milk quality payment program were collected for the determination of the SCC, the total bacterial count (TBC), the preliminary incubation count (PIC), the laboratory pasteurization count (PC), and the coliform count (CC). Based on the bulk tank SCC results, 63 herds were selected and categorized into three groups: (a) low SCC: ≤250,000 cells/mL (n = 16); (b) medium SCC: >250,000 ≤ 400,000 cells/mL (n = 24); and (c) high SCC: >400,000 cells/mL (n = 23). Socioeconomic profiles, attitudes, and knowledge of somatic cell count and milk quality were assessed using previously tested questionnaires, which were used to interview the herd managers of the 63 selected dairy herds. Among the findings, 87.0% of the dairy herds had an SCC < 400 × 10³ cells/mL, and presented milk production as the main economic activity of the farm, whereas only 47.0% of dairy herds with an SCC > 400 × 10³ cells/mL had milk production as the main economic activity of the farm (p < 0.031). In a total of 95% of the selected herds (n = 60), milking machines were used, with pipeline milking machines being more predominant, accounting for 70.8% in herds with a medium somatic cell count (SCC) and 78.3% in herds with a high SCC, as opposed to herds with a low SCC at 50% (p < 0.031). The frequency of dairy producers’ awareness of the maximum legal requirements for the bulk tank SCC and TBC was higher in herds with a higher SCC than in herds with a medium SCC. In conclusion, the results of this study indicate a significant association between the bulk tank somatic cell count (SCC) and dairy farmers’ knowledge about milk quality, mastitis control, and socioeconomic characteristics. These findings emphasize the importance of knowledge and the socioeconomic profiles of dairy farms in maintaining and enhancing milk quality in dairy herds. Full article

The fecundity characteristics and spawning strategy of Uroteuthis edulis in the East China Sea were investigated by observing the potential fecundity (PF), relative fecundity by dorsal mantle length (PF_ML) and relative fecundity by body weight (PF_BW). The relationship between fecundity and a single biological indicator was measured, and generalized additive models (GAMs) were fit by adding environmental variables to help better understand this comprehensive relationship. The long diameter and short diameter of the ovarian oocytes ranged from 0.72 mm to 4.74 mm and from 0.46 mm to 3.67 mm, respectively. The long and short diameters of oviducal eggs ranged from 0.61 mm to 5.12 mm and from 0.39 mm to 3.81 mm, respectively. The egg diameter had a unimodal distribution. The PF, PF_ML and PF_BW ranged from 540 to 13,743 cells, 5 to 86 cells/mm and 6 to 53 cells/g, respectively. Three fecundity indicators were unimodally distributed, and the PF_BW was more stable than the PF_ML (δ²PF_BW < δ²PF_ML). The fecundity and single biological indicators were fitted, and it was found that the PF and PF_ML were positively correlated with dorsal mantle length (ML) and body weight (BW). The generalized additive model (GAM) fitting showed that when considering the interaction between dorsal mantle length and sea surface height (M₁₃), the deviation explanation rate of the PF and PF_ML was the highest. Studies have shown that the ovary oocytes of U. edulis mature in batches, and then the eggs are laid in batches. The dorsal mantle length (ML), water temperature at 25 m depth (T₂₅) and sea surface height (SSH) are important indicators to measure the fecundity of U. edulis. These findings allow for a deeper understanding of the U. edulis population dynamics for the future management of this economically and ecologically important species. Full article

This study follows a previously defined framework to investigate the impact of impairment on performance in Para dressage athletes. Twenty-one elite Para dressage athletes (grades I to V) and eleven non-disabled dressage athletes (competing at Prix St. Georges or Grand Prix) participated. Data were collected in two phases: performing a two minute custom dressage test on a riding simulator while kinematic data were synchronously collected using inertial measurement units (2000 Hz) and optical motion capture (100 Hz), and clinically assessed using a battery of impairment assessment tools administered by qualified therapists. Impairment and performance measures were compared between Para and non-disabled athletes. Significant differences between athlete groups were found for all impairment measures and two performance measures: simulator trunk harmonics (p = 0.027) and athlete trunk dynamic symmetry (p < 0.001). Impairment assessments of sitting function and muscle tone could predict 19 to 35% of the impact of impairment on performance in Para athletes but not in non-disabled athletes. These findings provide the basis for a robust, scientific evidence base, which can be used to aid in the refinement of the current classification system for Para dressage, to ensure that it is in line with the International Paralympic Committee’s mandate for evidence-based systems of classification. Full article

The aims of this study were to examine the effect of luteotropic and luteolytic factors on the mRNA and protein expression of the coactivators HAT: histone acetyltransferase p300 (P300), cyclic adenosine monophosphate response element-binding protein (CREB), and steroid receptor coactivator-1 (SRC-1) and the corepressor: nuclear receptor corepressor-2 (NCOR-2) in bovine luteal cells on days 6–10 and 16–20. HAT and HDAC activities were also measured. The obtained results showed that luteotropic and luteolytic factors influence changes in the mRNA and protein levels of the coregulators of PGRs. However, they did not affect the activity of related HAT and HDAC, respectively. Therefore, it is possible that these factors, through changes in the expression of nuclear receptor coactivators and corepressors, may affect the functioning of the nuclear receptors, including PGRs, in the bovine CL. Full article

Unnatural diet composition and frequent feeding regimes may play an aetiological role in the multiple diseases prevalent in captive cheetahs. This study investigated the responses of captive-born (hand-reared) cheetahs (n = 6) to a reduced feeding frequency schedule distinguished by offering larger quantities of food less frequently. The study cheetahs were fed four once-daily meals per week during the 3-week treatment period, followed by a 3-week control period in which they were fed two daily rations six days a week. Total weekly food intake was maintained throughout the study. Variations in behaviour, faecal consistency score (FCS), and faecal glucocorticoid metabolite concentration were measured. Less frequent feeding resulted in higher FCS (p < 0.01) and locomotory behaviour (p < 0.05) among the studied cheetahs. Faecal glucocorticoid metabolite concentration demonstrated an initial acute stress response to the change in feeding frequency (p < 0.05) and subsequent adaptation. The results of the FCS analysis suggest that the more natural feeding pattern could have benefited the studied cheetahs’ gastrointestinal health without a significant behavioural or physiological stress response overall to the change in feeding frequency. Full article

We here provide the first comprehensive analysis and discussion on prey consumed by the European cave salamanders of the genus Speleomantes. Our study stems from the need to shed light on the still unknown foraging behavior adopted by Speleomantes cave salamanders. Starting from the published datasets on gut contents from all Speleomantes species (including hybrids), we here discuss additional information (i.e., species ecology, lower taxonomic level), which were systematically omitted from those data sets. We analyzed a data set consisting of 17,630 records from 49 categories of consumed prey recognized from gut contents of 2060 adults and juveniles Speleomantes. Flying prey accounted for more than 58% of the prey items, while elongated crawling prey accounted for no more than 16% of the diet within a single population. Among the total recognized prey items, only three can be surely ascribed to the group of strictly-cave species (i.e., troglobites), meaning that European cave salamanders mostly forage in surface environment, and therefore represent one of the major drivers of allochthonous organic matter in subterranean environments. Some of the consumed prey seemed to be aquatic, allowing us to hypothesize whether Speleomantes are able to catch prey from a shallow body water. Furthermore, European cave salamanders possess the ability to prey upon taxa characterized by particular anti-predator defenses, while morphological constraints seem to be the most important limit to prey consumption. For each specific case, we provide insights and propose hypotheses concerning the foraging behavior that need to be tested to properly understand the foraging behavior of this cryptic salamanders. Full article

Oxidative stress leads to a decrease in semen quality during semen cryopreservation and fresh semen production. Grape seed proanthocyanidins (GSPs) are endowed with well-recognized antioxidant, anti-inflammatory, anti-cancer, and anti-aging activities. Therefore, the objective of this experiment was to explore the effects of GSPs on the quality of fresh and cryopreserved semen to provide a basis for GSPs as a new dietary additive and semen diluent additive for males’ reproduction. Fresh semen from three healthy bulls aged 3 to 5 years old were gathered and mixed with semen diluents dissolved with 0 µg/mL, 30 µg/mL, 40 µg/mL, 50 µg/mL, and 60 µg/mL GSPs respectively. The motility, physiological structures (acrosome integrity, membrane integrity, mitochondrial activity), and antioxidant capacity of frozen–thawed sperm were measured after storage in liquid nitrogen for 7 days (d). Bulls were fed with 20 mg/kg body weight (BW) GSPs in their diet for 60 days; the weight of the bull is about 600 kg. Then, the reproductive performance and antioxidant indexes of bulls were measured before and after feeding. The results demonstrated that GSPs supplementation significantly increased sperm motility, physiological structures, GSH-Px, and CAT enzyme activities and significantly decreased MDA content in sperm during semen cryopreservation. The optimal concentration of GSPs was 40 µg/mL (p < 0.05). After 20 mg/kg (body weight) GSP supplementation, sperm motility was significantly heightened (p < 0.05), the sperm deformity rate was significantly reduced (p < 0.05), and antioxidant enzyme activities (such as SOD, CAT, and GSH-Px) were significantly enhanced (p < 0.05), and the production of MDA was significantly suppressed (p < 0.05) in serum compared with that before feeding. In conclusion, these results reveal that a certain concentration of GSPs has a good protective effect on sperm damage caused by semen cryopreservation and the reproductive performance reduction caused by stress in bulls, which may be attributed to the antioxidant function of GSPs. In summary, GSPs are a useful cryoprotective adjuvant and dietary additive for bull sperm quality. Full article

Fatigue-related subchondral bone injuries of the third metacarpal/metatarsal (McIII/MtIII) bones are common causes of wastage, and they are welfare concerns in racehorses. A better understanding of bone health and strength would improve animal welfare and be of benefit for the racing industry. The porosity index (PI) is an indirect measure of osseous pore size and number in bones, and it is therefore an interesting indicator of bone strength. MRI of compact bone using traditional methods, even with short echo times, fail to generate enough signal to assess bone architecture as water protons are tightly bound. Ultra-short echo time (UTE) sequences aim to increase the amount of signal detected in equine McIII/MtIII condyles. Cadaver specimens were imaged using a novel dual-echo UTE MRI technique, and PI was calculated and validated against quantitative CT-derived bone mineral density (BMD) measures. BMD and PI are inversely correlated in equine distal Mc/MtIII bone, with a weak mean r value of −0.29. There is a statistically significant difference in r values between the forelimbs and hindlimbs. Further work is needed to assess how correlation patterns behave in different areas of bone and to evaluate PI in horses with and without clinically relevant stress injuries. Full article