Animals

15 pages, 2945 KiB

Open AccessFeature PaperArticle

Morphology, Glycan Pattern, Heat Shock Proteins, and Sex Steroid Receptors Expression in the Tubal Fimbria Epithelium of the Baboon Papio hamadryas during the Menstrual Cycle

by Salvatore Desantis, Mario Cinone, Luca Lacitignola, Pietro Laricchiuta, Roberta Rossi, Antonio Ciro Guaricci, Leonardo Resta and Maria Albrizio

Animals 2024, 14(16), 2321; https://doi.org/10.3390/ani14162321 (registering DOI) - 11 Aug 2024

Abstract

The oviductal fimbria is the first extraovarian anatomical structure that the cumulus–oocyte complex (COC) encounters, and is sensitive to sex hormone changes. The morphology, glycan pattern, expression of heat shock proteins (HSPs), estradiol receptor (ER), and progesterone receptor (PR) were investigated in the [...] Read more.

The oviductal fimbria is the first extraovarian anatomical structure that the cumulus–oocyte complex (COC) encounters, and is sensitive to sex hormone changes. The morphology, glycan pattern, expression of heat shock proteins (HSPs), estradiol receptor (ER), and progesterone receptor (PR) were investigated in the oviductal fimbria epithelium of the baboon (Papio hamadryas) during the menstrual cycle. The morphology was investigated by light and scanning electron microscopy; the glycopattern was characterized using conventional and lectin histochemistry; HSPs (60, −70, −90), ER, and PR were localized immunohistochemically. Well-differentiated ciliated and nonciliated cells were present only during the preovulatory phase. The nonciliated cells contained small apical protrusions and thin microvilli. During the preovulatory phase (1) the luminal surface of the fimbria displayed acidic glycans, complex N-glycans containing fucose, and oligolactosamine residues; (2) nonciliated cells expressed HSP60 and HSP90 in the apical blebs, HSP70 in the nucleus and cytoplasm, as well as nuclear ERα and PR; (3) ciliated cells showed HSP70 in the nucleus, cytoplasm, and cilia that also expressed HSP90 and PR. These results are related to the function of the fimbria where the early COC–oviduct crosstalk occurs and may represent a benchmark for translational studies of other primates. Full article

(This article belongs to the Section Animal Reproduction)

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Figure 1
Macroscopic (A) and histological view (B) of baboon Papio hamadryas oviductal fimbriae stained with Hematoxylin-Eosin. In B, note the opening of the infundibulum (oi). oi, ostium of the infundibulum; mi, muscular of the infundibulum; arrow, mucosal folds of the fimbriae. Scale bar: (A) 2 mm; (B) 500 µm. Full article ">Figure 2
Light micrographs showing the morphological changes in the epithelium of baboon Papio hamadryas oviductal fimbriae during the menstrual cycle. Hematoxylin-Eosin staining. Scale bar: 10 µm. Full article ">Figure 3
Scanning electron micrographs of the epithelium of baboon Papio hamadryas oviductal fimbriae during the menstrual cycle. ci, cilia. Scale bar: 8 µm. Full article ">Figure 4
Conventional histochemical staining of fimbria epithelium of baboon Papio hamadryas oviduct during the menstrual cycle. Note the absence of PAS staining (A–C) and the presence of AB 2.5 staining (azur staining) during the follicular and preovulatory phase (D,E). HID staining (brown staining) was detected only during the preovulatory phase (E). In (A–C), nuclei were stained with Hematoxylin. In (D–F), nuclei were stained with nuclear fast red. lp, lamina propria; arrow, luminal surface of the epithelium; asterisk, blood vessel. Scale bar: 20 µm. Full article ">Figure 5
AAL, GNL, LCA, and RCA120 binding sites in the mucosal epithelium of baboon Papio hamadryas oviductal fimbriae during the menstrual cycle. (A–C), expression of aL-Fuc terminating glycans revealed with LTA. (D–F), presence of terminal α1-3mannose residue detected by GNL. (G–I), localization of complex N-linked glycans by means of LCA. (J–L), lactosamine terminating glycans identified by RCA120. Note that the investigated glycans were mainly expressed during the preovulatory phase compared to other menstrual cycle phases. bv, blood vessel; e, epithelium; lp, lamina propria; arrow, luminal surface of the epithelium. Scale bar: 40 µm. AAL, GNL, and LCA were FITC-conjugated lectins. RCA was TRITC-conjugated lectin. Full article ">Figure 6
Immunostaining pattern of the HSP60 (A–C), 70 (D–F), 90 (G–I) in the mucosal epithelium of baboon Papio hamadryas oviductal fimbriae during the menstrual cycle. The inset images display the absence of immunoreactivity in negative controls. cc, ciliated cell; e, epithelium; lp, lamina propria; nc, nonciliated cell; arrowhead, cilia; *, apical bleb. Scale bar: 20 µm; insets, 10 µm. Full article ">Figure 7
ERα and PR immunostaining pattern in the baboon Papio hamadryas oviductal fimbriae during the menstrual cycle. (A–C), ERα was present only in the nuclei. (D–F), PR immunoreactivity was observed unevenly in the cytoplasm and nucleus of epithelial cells during the follicular and luteal phases, whereas it was present in the nuclei of the nonciliated cells and in the cilia during the preovulatory phase. The inset images display the absence of immunoreactivity in negative controls. cc, ciliated cell; e, epithelium; lp, lamina propria; nc, nonciliated cell; arrowhead, cilia; asterisk, negative nucleus. Scale bar: 10 µm. Full article ">

12 pages, 6042 KiB

Open AccessArticle

Histological Characteristics of Follicles, Reproductive Hormones and Transcriptomic Analysis of White King Pigeon Illuminated with Red Light

by Ying Wang, Kui Zuo, Chi Zhang, Dongzhi Miao, Jing Chen, Haiming Yang and Zhiyue Wang

Animals 2024, 14(16), 2320; https://doi.org/10.3390/ani14162320 (registering DOI) - 10 Aug 2024

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Abstract

Red light (RL) has been observed to enhance egg production in pigeons, yet the underlying histological characteristics and molecular mechanisms remain less understood. This study included fifty-four pigeons to assess follicular histology, reproductive hormones, and ovarian transcriptomics on the third day of the [...] Read more.

Red light (RL) has been observed to enhance egg production in pigeons, yet the underlying histological characteristics and molecular mechanisms remain less understood. This study included fifty-four pigeons to assess follicular histology, reproductive hormones, and ovarian transcriptomics on the third day of the laying interval under RL and white light (WL). The results showed that the granulosa cell layer was significantly thicker under RL (p < 0.05), whereas the theca cell and connective tissue layers showed no significant differences (p > 0.05). Higher plasma estradiol (E₂) levels were recorded in the RL group (p < 0.05), as well as follicle stimulating hormone (FSH), although progesterone (P₄) levels were higher under WL (p < 0.05). Moreover, P₄ concentrations in follicle yolk significantly decreased under RL (p < 0.01), with higher FSH and E₂ levels in F1 yolk and similar increases in SF1 yolk (p < 0.01). Transcriptomic analysis revealed 4991 differentially expressed genes in the pigeon ovary. The protein–protein interaction network highlighted genes like HSD11B1, VEGFD, WNT6, SMAD6, and LGR5 as potential contributors to hierarchical follicle selection under RL. This research provides new insights into the molecular basis by which RL may promote hierarchical follicle selection and improve egg production in pigeons. Full article

(This article belongs to the Section Birds)

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Histological characteristics of pigeon follicles under red light. (A) F1 follicle under RL. (B) F1 follicle under WL. (C) F2 follicle under RL. (D) SF1 follicle under RL. (E) SF1 under WL. Full article ">Figure 2
The concentration of estradiol (E2, df = 16, A), progesterone (P4, df = 16, B) and follicle-stimulating hormone (FSH, df = 9, C) in pigeon plasma under red light. Values marked with different capital letters on the bars are extremely significantly different (p < 0.01). Full article ">Figure 3
The concentration of estradiol (E2, df = 4, A), progesterone (P4, df = 4, B) and follicle-stimulating hormone (FSH, df = 4, C) in pigeon follicle yolk under red light. Values marked with different capital letters on the bars are extremely significantly different (p < 0.01). Full article ">Figure 4
The volcano plot of differentially expressed genes. Full article ">Figure 5
GO enrichment analysis of differentially expressed genes. Full article ">Figure 6
KEGG enrichment analysis of differentially expressed genes. Full article ">Figure 7
Protein–protein interaction (PPI) network of differentially expressed genes. Full article ">Figure 8
The validation of RNA-Seq using RT-qPCR. Full article ">

15 pages, 5173 KiB

Open AccessArticle

A Comparative Analysis of the Gene Expression Profiles in the Mammary Glands of Lactating and Nonlactating Mares at the Second Month of Gestation

by Tseweendolmaa Ulaangerel, Min Wang, Bilig Zhao, Minna Yi, Yingchao Shen, Yibeeltu Mengkh, Xin Wen, Manglai Dugarjav and Gerelchimeg Bou

Animals 2024, 14(16), 2319; https://doi.org/10.3390/ani14162319 (registering DOI) - 9 Aug 2024

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Abstract

To investigate molecular regulation involved in lactation during pregnancy, this study focused on the transcriptomic profiles of mammary tissue from lactating and non-lactating Mongolian mares at the second month of gestation. A total of 4197 differentially expressed genes were identified by comparing mammary [...] Read more.

To investigate molecular regulation involved in lactation during pregnancy, this study focused on the transcriptomic profiles of mammary tissue from lactating and non-lactating Mongolian mares at the second month of gestation. A total of 4197 differentially expressed genes were identified by comparing mammary tissues from pregnant mares at two different states, including 1974 differentially expressed genes such as the milk protein-related genes a-s1-casein (CSN1S1), k-casein (CSN3), lactalbumin (LALBA), and lactoferrin (LTF), which were highly expressed in the lactating mares group, and overall, these differentially expressed genes were mainly associated with biological processes such as endoplasmic reticulum protein processing, the Toll-like receptor signaling pathway, steroid biosynthesis, cytokine‒cytokine receptor interactions, and amino sugar and nucleotide glycolysis. These findings serve as a foundation for investigating the molecular underpinnings of lactation in pregnant equids. Full article

(This article belongs to the Section Equids)

18 pages, 23890 KiB

Open AccessArticle

Exploration of Response Mechanisms in the Gills of Pacific Oyster (Crassostrea gigas) to Cadmium Exposure through Integrative Metabolomic and Transcriptomic Analyses

by Luyao Dong, Yanan Sun, Muyang Chu, Yuxin Xie, Pinyi Wang, Bin Li, Zan Li, Xiaohui Xu, Yanwei Feng, Guohua Sun, Zhongping Wang, Cuiju Cui, Weijun Wang and Jianmin Yang

Animals 2024, 14(16), 2318; https://doi.org/10.3390/ani14162318 (registering DOI) - 9 Aug 2024

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Abstract

Marine mollusks, including oysters, are highly tolerant to high levels of cadmium (Cd), but the molecular mechanisms underlying their molecular response to acute Cd exposure remain unclear. In this study, the Pacific oyster Crassostrea gigas was used as a biological model, exposed to [...] Read more.

Marine mollusks, including oysters, are highly tolerant to high levels of cadmium (Cd), but the molecular mechanisms underlying their molecular response to acute Cd exposure remain unclear. In this study, the Pacific oyster Crassostrea gigas was used as a biological model, exposed to acute Cd stress for 96 h. Transcriptomic analyses of their gills were performed, and metabolomic analyses further validated these results. In our study, a total of 111 differentially expressed metabolites (DEMs) and 2108 differentially expressed genes (DEGs) were identified under acute Cd exposure. Further analyses revealed alterations in key genes and metabolic pathways associated with heavy metal stress response. Cd exposure triggered physiological and metabolic responses in oysters, including enhanced oxidative stress and disturbances in energy metabolism, and these changes revealed the biological response of oysters to acute Cd stress. Moreover, oysters could effectively enhance the tolerance and detoxification ability to acute Cd exposure through activating ABC transporters, enhancing glutathione metabolism and sulfur relay system in gill cells, and regulating energy metabolism. This study reveals the molecular mechanism of acute Cd stress in oysters and explores the molecular mechanism of high tolerance to Cd in oysters by using combined metabolomics and transcriptome analysis. Full article

(This article belongs to the Topic Anthropic Impacts in Marine Coastal Waters: Assessment, Case Studies and Solutions)

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Biochemical alterations of the acute Cd exposure C. gigas. (a) Increased intracellular Cd level in Cd-exposed oyster individuals. The bar chart depicts mean levels and standard deviation (SD) values, with n = 3. Significance differences were determined by t-test (p < 0.05), denoted by asterisks (*). (b) The survival rate of oysters after 96 h of acute Cd exposure. (c–f) After Cd exposure, oxidative stress markers such as SOD (c), CAT (d), MDA (e), and GPx (f) were measured. Data were presented as the mean ± SD (n = 3). Distinguishing letters were assigned to indicate significant differences (p < 0.05). Star plots depict the impact of Cd exposure on gill samples. Full article ">Figure 2
Analysis of oyster transcriptome following a 96-hour exposure to Cd. (a) A volcano plot of DEGs is graphically represented with upregulated genes in red and downregulated genes in blue. (b) Hierarchical clustering based on the DEGs, where red signifies upregulation and blue signifies downregulation. (c) Enrichment of DEGs in GO terms categorized into cellular components, biological processes, and molecular functions. (d) KEGG pathway enrichment analysis of DEGs, with colors indicating p-value significance and bubble size reflecting the count of enriched genes. Full article ">Figure 3
Analysis of oyster metabolomes following a 96-hour exposure to Cd. (a) OPLS-DA score plot of the metabolomic data. (b) PLS-DA sorting test plot of the metabolomic data. (c) Hierarchical clustering was performed using 111 DEMs, with red indicating upregulation and blue representing downregulation, respectively. (d) Sample comparisons for the matchstick diagram. The top 20 metabolites of up and down are displayed in the matchstick diagram. The x-axis of the matchstick diagram represents log2 (Fold Change) values, the y-axis represents metabolites, and the size of the points corresponds to VIP values. The metabolites that are upregulated and downregulated are represented by the red and blue points, respectively. Full article ">Figure 4
Pathway analysis of the DEMs. (a) Top 20 KEGG pathway results. p-values are represented by colors, while pathway impact is indicated by the size of the bubbles. (b) KEGG regulatory network diagram. Red circles represent individual metabolic pathways, yellow circles depict enzyme information related to specific substances, green circles indicate background substances for a metabolic pathway, purple circles represent information on molecular modules of a certain substance category, blue circles represent chemical interactions involving a specific substance, and green squares denote differentially expressed substances identified in this comparison. Full article ">Figure 5
Correlation heatmap analysis of DEGs and DEMs. The DEMs are depicted on the y-axis, while the x-axis displays the DEGs. A correlation coefficient less than 0 is indicative of a negative correlation, and a coefficient greater than 0 indicates a positive correlation. Negative correlations are symbolized by the color blue, while positive correlations are represented by the color red. Full article ">Figure 6
Pathway enrichment analysis of DEGs and DEMs. (a) Bubble plot of KEGG enrichment for DEGs and DEMs. The x-axis represents the ratio of the number of enriched differential metabolites or genes annotated to metabolites or genes in the pathway to the total number in that pathway (ratio). The y-axis represents KEGG pathways jointly enriched in the metabolome and transcriptome. The count indicates the number of enriched metabolites or genes in the pathway. The colors represent p-values, with brighter colors indicating smaller p-values and more significant pathway enrichment. (b) iPath pathway map of shared enriched pathways. Colored boxes represent enriched pathways, nodes depict various biochemical molecules, lines represent biochemical reactions, and blue lines within the pathways indicate pathways jointly enriched with DEGs and DEMs. Full article ">Figure 7
Main biological pathway responses to acute Cd exposure in oysters. The blue boxes represent four core Cd exposure response pathways. The orange boxes represent DEMs associated with the main pathways. The green boxes represent downregulated DEGs related to the main pathways. The red elliptical frames represent upregulated DEGs associated with the main pathways. Full article ">

17 pages, 784 KiB

Open AccessArticle

Isolation of Aerobic Bacterial Species Associated with Palpable Udder Defects in Non-Dairy Ewes

by Mandefrot M. Zeleke, Paul R. Kenyon, Kate J. Flay, Danielle Aberdein, Sarah J. Pain, Niluka Velathanthiri and Anne L. Ridler

Animals 2024, 14(16), 2317; https://doi.org/10.3390/ani14162317 - 9 Aug 2024

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Abstract

The objectives of these studies were to identify associations between udder half defects (hard or lump) and bacteria isolated from milk or mammary tissue swabs, to compare with samples from normal udder halves at different physiological time points and to compare bacterial species [...] Read more.

The objectives of these studies were to identify associations between udder half defects (hard or lump) and bacteria isolated from milk or mammary tissue swabs, to compare with samples from normal udder halves at different physiological time points and to compare bacterial species isolated via milk and swabs of mammary tissue from within the same udder halves. A total of 1054 samples were aseptically collected from each udder half of 199 non-dairy breed (Romney) ewes from three different studies (Study A, n = 77; Study B, n = 74; and Study C, n = 48). Conventional bacterial culture and MALDI-ToF mass spectrometry were used for bacterial identification. Of the 225 samples from which bacteria were isolated, Mannheimia haemolytica and Streptococcus uberis were the dominantly identified species from defective udder halves, whereas coagulase-negative staphylococcus (CNS) species, mostly Staphylococcus simulans and Staphylococcus chromogenes, were more frequently isolated from normal udder halves. The ongoing presence of bacterial species over time was variable, although less frequently identified species showed less stability over time. A very high agreement (91.5%) of bacterial species identified was observed between the mammary tissue swab and udder half milk samples during post-weaning. In summary, palpable udder half defects were associated with bacterial positivity, and the ongoing presence of the bacteria over time was dependent on the species involved. Hence, culling ewes with palpable udder half defects that had more stable bacterial species could contribute to reducing the recurrence of palpable defects or mastitis. Full article

(This article belongs to the Section Small Ruminants)

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10 pages, 1139 KiB

Open AccessArticle

Adaptive Evolution and Functional Differentiation of Testis-Expressed Genes in Theria

by Yukako Katsura, Shuji Shigenobu and Yoko Satta

Animals 2024, 14(16), 2316; https://doi.org/10.3390/ani14162316 - 9 Aug 2024

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Abstract

Gene expression patterns differ in different tissues, and the expression pattern of genes in the mammalian testis is known to be extremely variable in different species. To clarify how the testis transcriptomic pattern has evolved in particular species, we examined the evolution of [...] Read more.

Gene expression patterns differ in different tissues, and the expression pattern of genes in the mammalian testis is known to be extremely variable in different species. To clarify how the testis transcriptomic pattern has evolved in particular species, we examined the evolution of the adult testis transcriptome in Theria using 10 species: two marsupials (opossum and Tasmanian devil), six eutherian (placental) mammals (human, chimpanzee, bonobo, gorilla, rhesus macaque, and mouse), and two outgroup species (platypus and chicken). We show that 22 testis-expressed genes are marsupial-specific, suggesting their acquisition in the stem lineage of marsupials after the divergence from eutherians. Despite the time length of the eutherian stem lineage being similar to that of the marsupial lineage, acquisition of testis-expressed genes was not found in the stem lineage of eutherians; rather, their expression patterns differed by species, suggesting rapid gene evolution in the eutherian ancestors. Fifteen testis-expressed genes are therian-specific, and for three of these genes, the evolutionary tempo is markedly faster in eutherians than in marsupials. Our phylogenetic analysis of Rho GTPase-activating protein 28 (ARHGAP28) suggests the adaptive evolution of this gene in the eutherians, probably together with the expression pattern differentiation. Full article

(This article belongs to the Special Issue Recent Advances in Genetic Mechanism and Evolution of Sex Determination and Chromosomes)

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Figure 1
Acquisition/loss of testis-expressed genes in 10 species. The genes at each clade are listed in <a href="#animals-14-02316-t001" class="html-table">Table 1</a>, <a href="#animals-14-02316-t002" class="html-table">Table 2</a>, <a href="#app1-animals-14-02316" class="html-app">Tables S2 and S3</a>, and the left and right of the ‘/’ symbols mean acquisition and loss of testis-expressed genes, respectively. Full article ">Figure 2
The phylogenetic trees, branch lengths, and dn/ds ratios for three therian-specific testis genes in 10 species. (A) ARHGAP28 genes (1833 bp). (B) SYNM genes (2235 bp). (C) PDZRN3 genes (2256 bp). The trees were constructed by the NJ method using the number of nucleotide differences. The average and standard deviation of branch lengths in six eutherians are shown above the black bold branch, and those in two marsupials are shown above the gray bold branch. The dn/ds ratio in six eutherian or two marsupial pairs is shown under the branch. ** and * mean p < 0.01 and p < 0.05, respectively, and are supported by Fisher’s exact test. Full article ">Figure 2 Cont.
The phylogenetic trees, branch lengths, and dn/ds ratios for three therian-specific testis genes in 10 species. (A) ARHGAP28 genes (1833 bp). (B) SYNM genes (2235 bp). (C) PDZRN3 genes (2256 bp). The trees were constructed by the NJ method using the number of nucleotide differences. The average and standard deviation of branch lengths in six eutherians are shown above the black bold branch, and those in two marsupials are shown above the gray bold branch. The dn/ds ratio in six eutherian or two marsupial pairs is shown under the branch. ** and * mean p < 0.01 and p < 0.05, respectively, and are supported by Fisher’s exact test. Full article ">

10 pages, 1792 KiB

Open AccessCase Report

Reconstruction of the Quadriceps Extensor Mechanism with a Calcaneal Tendon–Bone Allograft in a Dog with a Resorbed Tibial Tuberosity Fracture

by Hyunho Kim, Haebeom Lee, Daniel D. Lewis, Jaemin Jeong, Gyumin Kim and Youngjin Jeon

Animals 2024, 14(16), 2315; https://doi.org/10.3390/ani14162315 - 9 Aug 2024

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Abstract

A non-reducible tibial tuberosity fracture is a rare complication of tibial tuberosity transposition performed during correcting of medial patella luxation (MPL) in dogs. This condition severely disrupts the quadriceps extensor mechanism, leading to significant pelvic limb lameness. An 11-year-old, 1.8 kg spayed female [...] Read more.

A non-reducible tibial tuberosity fracture is a rare complication of tibial tuberosity transposition performed during correcting of medial patella luxation (MPL) in dogs. This condition severely disrupts the quadriceps extensor mechanism, leading to significant pelvic limb lameness. An 11-year-old, 1.8 kg spayed female Yorkshire Terrier sustained a comminuted left tibial tuberosity fracture during surgical correction of an MPL. Six months after surgery, the dog was markedly lame and unable to extend the left stifle. Radiographs revealed patella alta and resorption of the fragmented tibial tuberosity. A composite frozen allogeneic calcaneal tendon–bone block was utilized to reconstruct the tibial tuberosity and reattach the patellar ligament. Initial postoperative radiographs confirmed restoration of a normal patellar ligament to patella length ratio (1.42). Both the allogeneic bone used for tibial tuberosity reconstruction and the tendon used to reattach the patellar ligament were successfully integrated. The dog regained satisfactory limb function without recurrence of patella luxation, as reported by the owners 29 months postoperatively. The use of a calcaneal tendon–bone allograft effectively restored the functional integrity of the quadriceps extensor mechanism, providing a viable option for addressing quadriceps insufficiency resulting from the loss of the osseous tibial insertion. Full article

(This article belongs to the Special Issue Small Animal Orthopedic Surgery, Physical Therapy and Rehabilitation)

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Figure 1
(A) Craniocaudal and (B) mediolateral radiographic views of the left stifle taken six months following initial surgery to address a grade III/IV medial patellar luxation. The images display tibial tuberosity absorption (white arrow) and proximal dislocation of the patella (arrowhead). Additionally, an ossicle (approximately 5 mm × 1.5 mm) with reduced bone density cranial to the infrapatellar fat pad was observed (yellow arrow). Full article ">Figure 2
Schematic diagrams illustrating the surgical technique. (A) The osseous segment of the calcaneal tendon–bone allograft is secured within the prepared tibial bone bed using two Kirschner wires. A non-absorbable suture is placed through the proximal tibia, patellar ligament, and proximal patellar tendon. (B) Craniocaudal and (C) mediolateral images show that the allogenic tendon is augmented to enhance both the patellar ligament and the quadriceps tendon. Additionally, an encircling patellar suture is placed as a supplemental restraint to medial luxation. Full article ">Figure 3
Intraoperative gross photographs. (A) Debrided bone displaying multiple holes for osteostixis. A noticeable gap is observed between the distal patellar ligament and the tibial bed (arrow). (B) The recombinant human bone morphogenetic protein-2 with hydroxyapatite applied to the bone surface (arrow). (C) Allogenic calcaneal bone block secured to the tibial bed using Kirschner wires (arrow). (D) A Krackow suture (arrow) initiated at the medial margin of the patellar ligament, encircling the proximal patella, and terminating at the lateral margin. (E) Two free ends of the suture (arrow) exit at the calcaneal bone–tendon allograft junction. (F) Both ends of the suture are threaded through the tibia and anchored with a suture button (arrow). (G) The grafted tendon is affixed to the patellar ligament and quadriceps tendon using multiple simple interrupted sutures (arrow). (H) Gross morphology of the completed implantation of the allograft. FS: femur side; TS: tibia side; Me: medial; La: lateral. Full article ">Figure 4
Immediate postoperative craniocaudal (A) and mediolateral (B) radiographic images, alongside sequential postoperative mediolateral radiographic images at immediate (C), 4 weeks (D), 12 weeks (E), and 28 weeks (F) post-surgery, with a PLL (marked as L in (C)): PL (marked as P in (C)) of 1.42, 1.46, 1.54 and 1.54, respectively. The patella remains normally positioned within the trochlear groove across all stages. The reconstruction of the tibial tuberosity using a calcaneus bone block allograft is distinctly visible. Due to skin irritation caused by the protruding ends of the Kirschner wires, pins and tension band wiring were removed at 26 weeks post-surgery. PLL, patellar ligament length; PL, patella length. Full article ">

11 pages, 5314 KiB

Open AccessArticle

Identification of ActivinβA and Gonadotropin Regulation of the Activin System in the Ovary of Chinese Sturgeon Acipenser sinensis

by Huamei Yue, Huan Ye, Rui Ruan, Hao Du and Chuangju Li

Animals 2024, 14(16), 2314; https://doi.org/10.3390/ani14162314 - 9 Aug 2024

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Abstract

Activin is a dimeric growth factor with diverse biological activities in vertebrates. This study aimed to investigate the regulatory role of the activin signaling pathway in the ovary of the endangered, cultured sturgeon species Acipenser sinensis. One activinβA subunit was identified, with [...] Read more.

Activin is a dimeric growth factor with diverse biological activities in vertebrates. This study aimed to investigate the regulatory role of the activin signaling pathway in the ovary of the endangered, cultured sturgeon species Acipenser sinensis. One activinβA subunit was identified, with a full-length complementary DNA (cDNA) sequence of 1572 base pairs. Multiple sequence alignment suggested that ActivinβA shared high sequence identities with its counterparts in four other sturgeon species. Phylogenetic analysis indicated the conserved evolution of ActivinβA among vertebrates from mammals to fish species. Transcripts of activinβA were distributed ubiquitously in the liver, kidney, intestine, ovary, midbrain, hypothalamus, and pituitary, with the highest transcription found in the pituitary. In Chinese sturgeon ovarian cells, in vitro human recombinant Activin A incubation stimulated the activin system-related gene transcriptions of activinβA, follistatin, its receptors -activinRIIA and activinRIIB, and drosophila mothers against decapentaplegic proteins (smads) smad2, smad3, and smad4. Ovary development-related mRNA levels of cyp19a1a and aromatase receptors of erα and erβ were enhanced by Activin A or human chorionic gonadotropin (hCG) incubation. Furthermore, 15 IU/mL hCG treatment increased the transcription levels of activinβA, follistatin, activinRIIA, and smad2. This suggested that the activin system was functional for the regulation of ovary development in Chinese sturgeon, possibly under the regulation of gonadotropin, by recruiting activinβA, follistatin, activinRIIA, and smad2. These results were helpful for the molecular exploration of activin signaling in fish species, as well as the ovarian maturation regulation of A. sinensis. Full article

(This article belongs to the Section Animal Physiology)

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Figure 1
Multiple amino acid sequence alignment of ActivinβA of Chinese sturgeon with other representative vertebrates. Identical and similar amino acids are highlighted with black and gray shading. Identical amino acids were further marked with asterisks. Sequence identities are indicated at right. Full article ">Figure 2
The Maximum Likelihood phylogenetic tree of Smad2/3 constructed by Mega X software of representative vertebrates. Horizontal branch lengths are proportional to estimated divergence of the sequence from the branch point. Full article ">Figure 3
Tissue distribution analysis of activinβA evaluated by relative real-time PCR. Data are normalized to ef1α mRNA and represent mean ± SD of three separate experiments. Values with different letters above are significantly different (p < 0.05). Full article ">Figure 4
Effect of human Activin A incubation on the transcriptions of activin, folistatin, activin receptors (A), smad genes (B), and ovary development-related genes (C) in the ovarian cells evaluated by relative real-time PCR. Data are normalized to ef1α mRNA and represent mean ± SD of three separate experiments. Asterisks denote significant difference from control at p < 0.05. Full article ">Figure 5
Effect of hCG incubation on the transcriptions of activin, follistatin, activin receptors (A), smad genes (B), and ovary development-related genes (C) in ovarian cells evaluated by relative real-time PCR. Data are normalized to ef1α mRNA and represent mean ± SD of three separate experiments. Asterisks denote significant difference from control at p < 0.05. Full article ">

18 pages, 2893 KiB

Open AccessArticle

Early Detection of Chronic Kidney Disease Using Plasma Neutrophil Gelatinase-Associated Lipocalin and Kidney Injury Molecule-1 in Small-Breed Dogs: A Retrospective Pilot Study

by Hyo-Sung Kim, Han-Jun Kim and Sun-Hee Do

Animals 2024, 14(16), 2313; https://doi.org/10.3390/ani14162313 - 9 Aug 2024

Viewed by 305

Abstract

Multiple diagnostic modalities are urgently needed to identify early-stage kidney diseases. Various molecules have been investigated; however, most studies have focused on identifying specific biomarkers in urine. Considering that assessing the symmetrical dimethylarginine (SDMA) plasma concentration is more suitable as an early diagnostic [...] Read more.

Multiple diagnostic modalities are urgently needed to identify early-stage kidney diseases. Various molecules have been investigated; however, most studies have focused on identifying specific biomarkers in urine. Considering that assessing the symmetrical dimethylarginine (SDMA) plasma concentration is more suitable as an early diagnostic test for chronic kidney disease (CKD) in routine veterinary practice, we aimed to investigate the clinical usefulness of plasma neutrophil gelatinase-associated lipocalin (pNGAL) and plasma kidney injury molecule-1 (pKIM-1) concentrations for CKD detection in small-breed dogs. Through a retrospective analysis, we found that numerous clinicopathological data showed a log-normal distribution, even when they satisfied normality tests. Moreover, the log-transformed pNGAL and pKIM-1 concentrations successfully identified CKD International Renal Interest Society (IRIS) stages 1–4 and the risk group with underlying CKD risk factors. Correlation analysis and group comparison of other factors confirmed the possibility of using these two biomarkers for detecting the CKD risk group and IRIS stage 1. Receiver operating characteristic curve analysis revealed that the diagnostic accuracy for discriminating the risk group was superior in the order of pKIM-1, pNGAL, SDMA, and serum creatinine levels. In conclusion, these results suggest that pKIM-1 and pNGAL are possible early or quantifiable markers of insignificant CKD or can be at least used as an adjunct with traditional indicators. Full article

(This article belongs to the Special Issue Advanced Biomarker Research in Animal Pathological States)

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Correlation between traditional and novel kidney injury biomarkers. Biomarkers showed a linear relationship with each other after the X and Y axes were displayed as log scales. Each scattered dot represents an individual case, colored by the chronic kidney disease stage. The line indicates simple linear regression, accompanied by small black dots describing 95% confidence intervals. Abbreviations: SLG, slope gradient; R2, coefficient of determination (R-squared), indicating the goodness-of-fit measure of the linear model; pNGAL, plasma neutrophil gelatinase-associated lipocalin; sCr, serum creatinine; SDMA, symmetrical dimethylarginine; IRIS, International Renal Interest Society. Full article ">Figure 2
Normality and log-normality analysis of novel kidney injury biomarkers. pNGAL and pKIM-1 showed a log-normal distribution. Q-Q plots of two objective biomarkers were compared before and after logarithmic transformation. Q-Q plots of all other data are presented in <a href="#app1-animals-14-02313" class="html-app">Supplementary Figure S2</a>. Red dotted lines indicate the identity line, whereas points forming a straight line indicate a suitable distribution. Abbreviations: pNGAL, plasma neutrophil gelatinase-associated lipocalin; pKIM-1, plasma kidney injury molecule-1. Full article ">Figure 3
Statistical analysis of differences in the concentrations of biomarkers between the groups after log transformation. No significant differences in serum creatinine (sCr) and symmetrical dimethylarginine (SDMA) concentrations were observed between the control, risk, and stage 1 groups, whereas a significant difference in pNGAL and pKIM-1 was found. The p-value between the stage 3–4 group and other groups was omitted because they were all <0.001 for four biomarkers. The lines indicate a mean ± 95% confidence interval. Full article ">Figure 4
Correlation matrix between kidney biomarkers and clinicopathological indicators. The numbers in each cross box are r coefficients, indicating linearity. Red and blue correspond to positive and negative correlations, respectively. Hashes (#) indicate that the markers were analyzed after log2 transformation, as they showed a log-normal distribution. The black superscripted “ns” indicates that the correlation was not statistically significant; otherwise, the p-value was <0.05. A white superscripted asterisk (*) indicates a comparison with sCr; *, p < 0.05. Abbreviations: BUN, blood urea nitrogen; sCr, serum creatinine; SDMA, symmetric dimethylarginine; pNGAL, plasma neutrophil gelatinase-associated lipocalin; pKIM-1, plasma kidney injury molecule-1; RBC, red blood cell; HCT, hematocrit; HGB, hemoglobin; RETIC, reticulocyte count; INF, inflammation; WBC, white blood cell; NEU, neutrophil count; LYM, lymphocyte count; MONO, monocyte count; CRP, C-reactive protein; TP, total protein; ALB, albumin; GLB, globulin; A/G, albumin-to-globulin ratio; ALT, alanine aminotransferase; AST, aspartate aminotransferase; ALP, alkaline phosphatase; AML, amylase; LPS, lipase; CK, creatinine kinase; CO, calculated osmolality; UP, urine protein concentration; UC, urine creatinine concentration; UPC, urine protein-to-urine creatinine ratio; USG, urine-specific gravity; BW, body weight; sysBP, systolic blood pressure; MMVD, myxomatous mitral valve disease; HAC, hyperadrenocorticism. Full article ">Figure 5
Accuracy of the biomarkers in detecting chronic kidney disease. (A) The control group was classified as the negative instance, whereas the risk and IRIS stage 1–4 groups were classified as positive instances. Overall, the areas under the curves of pNGAL and pKIM-1 were found to be comparable and were slightly higher than those of SDMA; sCr exhibited the lowest AUC. (B) The control and risk groups were classified as negative instances, whereas the IRIS stage 1–4 groups were classified as positive instances. Overall, the areas under the curves were highest in the order of pNGAL, symmetrical dimethylarginine (SDMA), pKIM-1, and serum creatinine (sCr). (C) The control, risk, and IRIS stage 1 groups were classified as negative instances, whereas the IRIS stage 2–4 groups were classified as positive instances. Overall, the areas under the curves of pKIM-1 were higher than those of pNGAL. Abbreviations: AUC, area under the curve; pNGAL, plasma neutrophil gelatinase-associated lipocalin; pKIM-1, plasma kidney injury molecule-1. Full article ">

20 pages, 6449 KiB

Open AccessArticle

Amur Tiger Individual Identification Based on the Improved InceptionResNetV2

by Ling Wu, Yongyi Jinma, Xinyang Wang, Feng Yang, Fu Xu, Xiaohui Cui and Qiao Sun

Animals 2024, 14(16), 2312; https://doi.org/10.3390/ani14162312 - 9 Aug 2024

Viewed by 250

Abstract

Accurate and intelligent identification of rare and endangered individuals of flagship wildlife species, such as Amur tiger (Panthera tigris altaica), is crucial for understanding population structure and distribution, thereby facilitating targeted conservation measures. However, many mathematical modeling methods, including deep learning [...] Read more.

Accurate and intelligent identification of rare and endangered individuals of flagship wildlife species, such as Amur tiger (Panthera tigris altaica), is crucial for understanding population structure and distribution, thereby facilitating targeted conservation measures. However, many mathematical modeling methods, including deep learning models, often yield unsatisfactory results. This paper proposes an individual recognition method for Amur tigers based on an improved InceptionResNetV2 model. Initially, the YOLOv5 model is employed to automatically detect and segment facial, left stripe, and right stripe areas from images of 107 individual Amur tigers, achieving a high average classification accuracy of 97.3%. By introducing a dropout layer and a dual-attention mechanism, we enhance the InceptionResNetV2 model to better capture the stripe features of individual tigers at various granularities and reduce overfitting during training. Experimental results demonstrate that our model outperforms other classic models, offering optimal recognition accuracy and ideal loss changes. The average recognition accuracy for different body part features is 95.36%, with left stripes achieving a peak accuracy of 99.37%. These results highlight the model’s excellent recognition capabilities. Our research provides a valuable and practical approach to the individual identification of rare and endangered animals, offering significant potential for improving conservation efforts. Full article

(This article belongs to the Special Issue Conservation and Evolution Biology of Endangered Animals)

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17 pages, 1558 KiB

Open AccessArticle

Associations between Ultrasonographically Diagnosed Lung Lesions, Clinical Parameters and Treatment Frequency in Veal Calves in an Austrian Fattening Farm

by Julia Hoffelner, Walter Peinhopf-Petz and Thomas Wittek

Animals 2024, 14(16), 2311; https://doi.org/10.3390/ani14162311 - 8 Aug 2024

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Abstract

This study evaluated the significance and predictive value of ultrasonographic and physical examination on arrival at an Austrian fattening farm. Treatment frequency and average daily weight gain (ADG) were related to physical and ultrasonographic examination results. Additionally, the effect of an intranasal vaccination [...] Read more.

This study evaluated the significance and predictive value of ultrasonographic and physical examination on arrival at an Austrian fattening farm. Treatment frequency and average daily weight gain (ADG) were related to physical and ultrasonographic examination results. Additionally, the effect of an intranasal vaccination in half of the examined calves was studied. The clinical and ultrasonographic health status 600 calves was recorded at the beginning and end of fattening. Half of the calves received an intranasal vaccination (Bovalto^® Respi Intranasal). Overall, 44.5% showed an abnormal respiratory scoring (RS) and 56.0% showed signs of respiratory diseases in transthoracic ultrasonography (TUS) at arrival on the farm. For both RS and TUS, a categorization between ILL and HEALTHY was conducted. Results showed lower ADG in ILL calves (RS median: 0.93 kg/d; TUS median: 0.96 kg/d) compared to HEALTHY calves (RS median: 1.01 kg/d; TUS median: 1.01 kg/d). The median ADG was lower in not treated and ILL calves (RS median 0.90 kg/d; TUS: 0.93 kg/d) compared to treated and ILL calves (RS median 1.01 kg/d; TUS: 1.02 kg/d). Vaccination did not affect growth performance or occurrence of ILL, though treatment frequency was lower in VAC calves (17.0% in NVAC; 11.3% in VAC). The implementation of examination protocols for respiratory diseases may have a positive impact on production parameters (e.g., treatment frequency and ADG). Full article

(This article belongs to the Section Cattle)

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Segmentation of calves’ lungs in five examination areas. L1 = lobus cranialis sinister, L2 = lobus caudalis sinister, R1 = lobus cranialis dexter + lobus medialis dexter, R2 = lobus accessories (accessible in slaughter examination exclusively), R3 = lobus caudalis dexter [<a href="#B18-animals-14-02311" class="html-bibr">18</a>,<a href="#B32-animals-14-02311" class="html-bibr">32</a>]. Full article ">Figure 2
Significant difference in average daily weight gain (ADG in kg/d) between clinically ILL (n = 267) and HEALTHY (n = 333) calves were detected in the first respiratory scoring (RS) at the beginning of fattening. Different letters (a,b) within the category (ILL or HEALTHY) are statistically different (p < 0.05). Full article ">Figure 3
Significant difference in average daily weight gain (ADG in kg/d) between ultrasonographically ILL (n = 336) and HEALTHY (n = 264) calves detected in the first transthoracic ultrasonography scoring (TUS1) at the beginning of fattening. Different letters (a,b) within the category (ILL or HEALTHY) are statistically different (p < 0.05). Full article ">Figure 4
Difference in average daily weight gain (ADG in kg/d) in treated (n = 73) and not treated (n = 194) calves detected in clinically ILL calves at first physical examination at the beginning of fattening. Different letters (a,b) within the category (treated or not treated) are statistically different (p < 0.05). Full article ">Figure 5
Difference in average daily weight gain (ADG in kg/d) between treated (n = 80) and not treated (n = 256) ultrasonographically ILL calves at first ultrasonographic examination at the beginning of fattening. Different letters (a,b) within the category (treated or not treated) are statistically different (p < 0.05). Full article ">

12 pages, 976 KiB

Open AccessArticle

Agreement of Pain Assessment Using the Short Form of the Canine Glasgow Composite Measure Pain Scale between Veterinary Students, Veterinary Nurses, Veterinary Surgeons, and ECVAA-Diplomates

by Mireia Marco-Martorell, Natalie Duffy, Miguel Martinez, Thomas Maddox and Katherine Robson

Animals 2024, 14(16), 2310; https://doi.org/10.3390/ani14162310 - 8 Aug 2024

Viewed by 547

Abstract

Several pain scoring systems have been validated to measure pain in dogs. However, pain may not be adequately assessed since these tools are associated with high-level inter-observer variation. The aim of this study is to evaluate the agreement of pain assessment using the [...] Read more.

Several pain scoring systems have been validated to measure pain in dogs. However, pain may not be adequately assessed since these tools are associated with high-level inter-observer variation. The aim of this study is to evaluate the agreement of pain assessment using the CMPS-SF between veterinary students, veterinary nurses, veterinary surgeons, and European College of Veterinary Anaesthesia and Analgesia (ECVAA) diplomates. Forty-five client-owned dogs presented to a teaching hospital were enrolled in this prospective, observational study. All dogs were pain-scored in vivo, while a video of the assessment was recorded and subsequently evaluated by twenty assessors, with five per group. Mean scores between groups were compared, and agreement within groups and agreement of the average scores between groups were assessed by calculating the intraclass correlation coefficient (ICC). The intervention point at which dogs were deemed to require additional analgesia was also evaluated. Overall agreement of pain assessment was poor (ICC = 0.494). Nurses had the best inter-observer agreement (ICC = 0.656), followed by ECVAA diplomates (ICC = 0.540), veterinary surgeons (ICC = 0.478), and veterinary students (ICC = 0.432). The best inter-group agreement was between veterinary surgeons and nurses (ICC = 0.951) and between ECVAA diplomates and nurses (ICC = 0.951). Students were more likely to determine that additional analgesia was required compared to other groups. Pain assessment is key for animal welfare, and training in this area should be reinforced to improve consistency. Full article

(This article belongs to the Special Issue Advances in Companion Animal Pain Assessment)

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9 pages, 1593 KiB

Open AccessArticle

Rapid Detection of Getah Virus Antibodies in Horses Using a Recombinant E2 Protein-Based Immunochromatographic Strip

by Dengke Zhong, Jiayang Zheng, Zhiyong Ma, Yan Wang and Jianchao Wei

Animals 2024, 14(16), 2309; https://doi.org/10.3390/ani14162309 - 8 Aug 2024

Viewed by 235

Abstract

The prevalence and impact of Getah virus (GETV) are significant concerns in China. GETV can infect a wide range of animals, including horses, pigs, sheep, cattle, birds, and humans, resulting in substantial losses in the livestock and agricultural industries. GETV infection can cause [...] Read more.

The prevalence and impact of Getah virus (GETV) are significant concerns in China. GETV can infect a wide range of animals, including horses, pigs, sheep, cattle, birds, and humans, resulting in substantial losses in the livestock and agricultural industries. GETV infection can cause the development of ulcers and inflammation in the mouth and gums of horses, which result in pain and discomfort and lead to symptoms such as reduced appetite, drooling, and difficulty chewing. As a result, there is a pressing need for efficient and rapid disease diagnosis methods. However, the currently available diagnostic methods have limitations in terms of operational time, equipment, and the experience of the individuals using them. In this study, a rapid, specific, and sensitive detection method was developed using a colloidal gold-based immunochromatographic strip (ICS) for the detection of antibodies against GETV in horses. To prepare the ICS, the antigen domain of the E2 glycoprotein of GETV was expressed using the Escherichia coli expression system after analysis with DNAstar v7.1 software. The nitrocellulose membrane was coated with rE2 protein or SPA to form the test line and control line, respectively. After optimizing the reaction conditions, the sensitivity, specificity, and repeatability of the strip were verified. The results showed that the test strip had a detection limit of up to 1:320 dilutions for GETV-positive serum, with no cross-reactivity observed with other equine-susceptible pathogens such as equine arteritis virus (EAV), equine herpesvirus-1 (EHV-I), equine infectious anemia virus (EIAV), equine influenza virus (EIV), African horse sickness virus (AHSV), and Japanese encephalitis virus (JEV). Furthermore, the ICS exhibited a concordance rate of 94.0% when testing 182 clinical serum samples compared to the virus neutralization test. Overall, this ICS diagnosis method will be an effective tool for the rapid detection of GETV in the field. Full article

(This article belongs to the Special Issue Pathogenesis, Immunology and Epidemiology of Veterinary Viruses)

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Schematic diagram of the immunochromatographic strip (ICS) test for detecting GETV antibodies in horse. (A) Structure of the ICS. (B) Interpretation of the results using ICS. A positive sample shows two red bands on the membrane strip; a negative sample shows only one band on the control line. If there is no colored band at all or there is only one colored band on the test line, the test is invalid. C, control line; T, test line; S, sampling hole. Full article ">Figure 2
Design, preparation, and identification of recombinant protein rE2. (A) Amino acid sequence of the E2 protein of Getah virus strain SH05-6 was obtained from GenBank (accession number: EU015066.1) and included three functional domains. Antigenic domains of Getah virus E2 glycoprotein were analyzed using the Protean program of DNAstar v7.1. Colored boxes indicate domains A, B, and C, which were predicted in reference to those of Chikungunya virus [<a href="#B22-animals-14-02309" class="html-bibr">22</a>]. (B) Production and identification of recombinant protein rE2.M: protein marker. Lane 1: Sodium dodecyl sulfate–polyacrylamide gel electrophoresis analysis of purified rE2 (33 kDa) expressed by vector pCold I. Lanes 2 and 4: Western blotting analysis of the E. coli-containing pCold using anti-His monoclonal antibody (Lane 2) and GETV-positive horse serum (Lane 4). Lanes 3 and 5: Western blotting analysis of the E. coli-containing pCold-E2 using anti-His monoclonal antibody (Lane 3) and GETV-positive horse serum (Lane 5). Full article ">Figure 3
Sensitivity and specificity testing of the ICS assay. (A) Sensitivity of the ICS. GETV-positive serum was diluted from 1:10 to 1:1280 to determine the sensitivity of the ICS. The detection limit of the test strip was up to 1:320 dilutions for the GETV-positive serum. Neg: negative horse serum. (B) Specificity of the ICS. Sera positive for different horse viruses (EHV1, EAV, EIV, EIAV, AHSV, and JEV) were used to evaluate the specificity of the ICS. The ICS did not cross-react with other viral-positive sera. Full article ">

14 pages, 2086 KiB

Open AccessArticle

Pet Owners’ Preferences for Quality of Life Improvements and Costs Related to Innovative Therapies in Feline Pain Associated with Osteoarthritis—A Quantitative Survey

by Andrea Wright, Edwina Gildea, Louise Longstaff, Danielle Riley, Nirav Nagda, Kristina DiPietrantonio, Ashley Enstone, Robin Wyn and David Bartram

Animals 2024, 14(16), 2308; https://doi.org/10.3390/ani14162308 - 8 Aug 2024

Viewed by 243

Abstract

This research aimed to explore UK cat owners’ preferences for treatments for feline osteoarthritis (OA) by exploring preferences around quality of life (QoL) improvements, safety considerations, and costs associated with hypothetical innovative pain therapies. Aspects identified in an existing conceptual framework were extracted [...] Read more.

This research aimed to explore UK cat owners’ preferences for treatments for feline osteoarthritis (OA) by exploring preferences around quality of life (QoL) improvements, safety considerations, and costs associated with hypothetical innovative pain therapies. Aspects identified in an existing conceptual framework were extracted for inclusion in exploratory interviews with cat owners (n = 3) to identify key domains that contribute to the QoL of cats. QoL descriptions for cats with OA and hypothetical product attributes were developed and validated through interviews with veterinarians (n = 3). An online survey was subsequently shared with 255 pet owners in the UK. Pet owners were presented with QoL descriptions and hypothetical product attributes to gather their preferences for QoL improvements and their willingness to pay (WTP) for (unbranded) pain therapies at various price points. Pet owners were motivated to improve their cats’ QoL, which translated into WTP for therapies; specifically, pet owners valued QoL improvements in mobility, pain expression, and well-being. When presented with a product profile of the hypothetical novel monoclonal antibody (mAb) and cost, 50% of cat owners were willing to pay more for a mAb that is expected to have improved efficacy and safety when compared to a hypothetical standard of care (SoC). Significantly more pet owners preferred the mAb than the SoC when price was not presented (p < 0.01), with product efficacy and safety driving pet owners’ decision-making. The majority of pet owners did not agree that taking their cats to the veterinarian once a month for their treatment would be burdensome. Cat owners in the UK are motivated to improve their cats’ QoL, which translates into WTP for the efficacious treatment of pain associated with osteoarthritis. Veterinarians should offer cat owners the pain treatment they feel is best suited for improving the cat’s QoL and to ensure subsequent owner-pet bond is preserved. Full article

(This article belongs to the Special Issue Behavior, Welfare, Health and Care of Aging Pets)

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16 pages, 1307 KiB

Open AccessArticle

Vibration as a New Survey Method for Spiders

by Rachael Harris, Robert Raven, Andrew Maxwell and Peter J. Murray

Animals 2024, 14(16), 2307; https://doi.org/10.3390/ani14162307 - 8 Aug 2024

Viewed by 415

Abstract

Spiders have important ecological roles as generalist predators, are a significant source of food for many other species, and are bioindicators of environmental health. However, spiders are poorly studied. Given their importance, a comparison of spider survey methods used to determine differences in [...] Read more.

Spiders have important ecological roles as generalist predators, are a significant source of food for many other species, and are bioindicators of environmental health. However, spiders are poorly studied. Given their importance, a comparison of spider survey methods used to determine differences in spider diversity and abundance is required to understand their limitations and biases. A new survey method to attract spiders, based on vibration from an idling diesel tractor, was tested and compared to the traditional methods of pitfall trapping and hand collection of spiders at night. Across the three survey methods, there were, in total, 2294 spiders in 34 families, 138 genera, and 226 species identified. Spider species diversity and richness were significantly greater for spiders collected at night than from the other two methods (spiders collected in pitfall traps and attracted to vibration). The collection of spiders using the night collection and vibration-based methods were very similar in terms of labor required and material costs. Of all spider species identified, 80% were captured during hand collection, 30% through pitfall trapping, and 30% from vibration-based collection. Most species of spiders caught in pitfall traps were species known to be primarily ground-dwelling, whereas both arboreal and ground-dwelling spiders were collected at night and as a result of being attracted and collected using the vibration-based method. Full article

(This article belongs to the Section Ecology and Conservation)

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10 pages, 649 KiB

Open AccessReview

Loco-Regional Anaesthesia during Standing Laparoscopic Ovariectomy in Equids: A Systematic Review (2003–2023) of the Literature

by Giada Giambrone, Giuseppe Catone, Gabriele Marino, Enrico Gugliandolo, Renato Miloro and Cecilia Vullo

Animals 2024, 14(16), 2306; https://doi.org/10.3390/ani14162306 - 8 Aug 2024

Viewed by 235

Abstract

Laparoscopic ovariectomy is generally performed with equids in the standing position, with the animals heavily sedated and restrained in stocks. This procedure may be quite painful, and it is essential first to manage intraoperative pain to complete the surgery, respecting the animal’s welfare [...] Read more.

Laparoscopic ovariectomy is generally performed with equids in the standing position, with the animals heavily sedated and restrained in stocks. This procedure may be quite painful, and it is essential first to manage intraoperative pain to complete the surgery, respecting the animal’s welfare and, at the same time, ensuring the safety of the operators. Laparoscopy requires multiple small incisions to introduce the instruments, with one to two incisions enlarged sufficiently to remove the ovary. The surgical procedure must be associated with effective pain control, usually obtained with loco-regional anaesthesia, mesovarian injection, mesovaric or ovarian topical anaesthesia, and epidural anaesthesia. This systematic review aims to discuss articles published from 2003 to 2023 on treating loco-regional anaesthesia in standing laparoscopic ovariectomy in association with an evaluation of pain. The literature review was undertaken according to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines on three databases (NCBI-PubMed, Web of Science, and SciVerse Scopus). Despite the collected papers numbering 36, we identified only five eligible papers, demonstrating that few studies are performed in order to evaluate the quality of analgesia with loco-regional anaesthesia in standing laparoscopic ovariectomy in equids. The authors of this systematic review agree that the association of injectable and epidural anaesthesia is the best solution to manage intraoperative pain in standing laparoscopic ovariectomy in equids. Full article

(This article belongs to the Special Issue Recent Advances in Animals Anesthesia, Analgesia and Pain Management during Gonadectomy)

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8 pages, 1745 KiB

Open AccessOpinion

The Silent Threat: Unraveling the Impact of Rabies in Herbivores in Brazil

by Marcelo Cardoso da Silva Ventura, Jéssica Milena Moura Neves, Randyson da Silva Pinheiro, Marcos Vinicius Costa Santos, Elba Regina Sampaio de Lemos and Marco Aurelio Pereira Horta

Animals 2024, 14(16), 2305; https://doi.org/10.3390/ani14162305 - 8 Aug 2024

Viewed by 291

Abstract

Rabies, a zoonotic viral disease, poses a significant threat due to its adaptability to diverse environments. Herbivore rabies, predominantly affecting cattle, horses, and goats in Brazil, remains a concern, results in substantial losses in the livestock industry, and poses risks to public health. [...] Read more.

Rabies, a zoonotic viral disease, poses a significant threat due to its adaptability to diverse environments. Herbivore rabies, predominantly affecting cattle, horses, and goats in Brazil, remains a concern, results in substantial losses in the livestock industry, and poses risks to public health. Rabies virus transmission, primarily through hematophagous bats in Latin America, underscores the need for effective strategies, and vaccination plays a crucial role in controlling herbivorous rabies, with systematic vaccination beingly the primary method. Efforts to control rabies in herbivores include vaccination campaigns, public awareness programs, and the enhancement of surveillance systems. Despite these initiatives, rabies persists and imposes an economic burden and a significant health risk. Economic impacts include losses in the livestock industry, trade restrictions on livestock products, and financial burdens on governments and farmers owing to control measures. Despite the considerable costs of campaigns, surveillance, and control, investing in rabies vaccination and control not only safeguards livestock, but also preserves public health, reduces human cases, and strengthens the sustainability of the livestock industry. Mitigating the impact of herbivorous rabies in Brazil requires integrated approaches and continuous investments in vaccination, surveillance, and control measures to protect public health and ensure the sustainability of the livestock industry, thus contributing to food and economic security. Full article

(This article belongs to the Special Issue Pathogens in the Wildlife–Livestock–Human Interface)

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14 pages, 308 KiB

Open AccessArticle

Carcass and Meat Characteristics of Cull Heifers from Different Genetic Groups Fed Diets with Different Sources of Nonprotein Nitrogen in Confinement

by Manoel Gustavo Paranhos da Silva, Luís Carlos Vinhas Ítavo, Camila Celeste Brandão Ferreira Ítavo, Marina de Nadai Bonin Gomes, Angelo Herbet Moreira Arcanjo, Jessika Rodrigues de Figueiredo Moura, Brenda Farias da Costa Leite Lopes, Lucimara Modesto Nonato and Rodrigo da Costa Gomes

Animals 2024, 14(16), 2304; https://doi.org/10.3390/ani14162304 - 8 Aug 2024

Viewed by 222

Abstract

The aim of this study was to evaluate the effect of genetic groups and diets with different sources of nonprotein nitrogen (NPN) on the carcass and meat characteristics of beef heifers. The meat from 40 heifers (20 ½ Angus ½ Nellore (A × [...] Read more.

The aim of this study was to evaluate the effect of genetic groups and diets with different sources of nonprotein nitrogen (NPN) on the carcass and meat characteristics of beef heifers. The meat from 40 heifers (20 ½ Angus ½ Nellore (A × N) and 20 ½ Charolais ½ Nellore (L × N)), finished in feedlots, was used. The heifers were fed diets containing different sources of NPN—(1) a diet with livestock urea and protected urea (LPU) and (2) a diet with extruded urea (EU)—in a completely randomized design with a 2 × 2 factorial arrangement. Carcass, composition and meat quality evaluations were carried out. There were no significant interactions between diet and genetic group for most of the variables evaluated (p > 0.05). The A × N heifers had higher hot carcass weights (305.73 vs. 279.80 kg), loin eye areas (80.87 vs. 75.45 cm²), subcutaneous fat thicknesses (8.69 vs. 6.35 mm) and lower shear forces (6.98 vs. 7.7 kg) compared to the C × N heifers (p < 0.05). The meat from the A × N heifers had higher proportions of saturated fatty acids (49.41 vs. 47.95%), with no effects on the proportions of monounsaturated (47.57%) and polyunsaturated (4.01%) fatty acids. The A × N heifers had better carcass and meat characteristics, while the C × N heifers had meat and fat with better fatty acid profiles. Full article

(This article belongs to the Special Issue Carcass Traits and Meat Quality in Cattle)

18 pages, 3149 KiB

Open AccessArticle

Gene Expression Analysis before and after the Pelvic Flexure in the Epithelium of the Equine Hindgut

by Cameron D. Moss, Amber L. Wilson, Kailee J. Reed, Kaysie J. Jennings, Isabelle G. Z. Kunz, Gabriele A. Landolt, Jessica Metcalf, Terry E. Engle and Stephen J. Coleman

Animals 2024, 14(16), 2303; https://doi.org/10.3390/ani14162303 - 8 Aug 2024

Viewed by 418

Abstract

Previous research demonstrated the distribution of distinct microbial communities in the equine hindgut surrounding the pelvic flexure. The current study evaluated gene expression in epithelial tissues surrounding the pelvic flexure to characterize patterns that might correlate with microbial distribution. Gene expression was determined [...] Read more.

Previous research demonstrated the distribution of distinct microbial communities in the equine hindgut surrounding the pelvic flexure. The current study evaluated gene expression in epithelial tissues surrounding the pelvic flexure to characterize patterns that might correlate with microbial distribution. Gene expression was determined by analyzing RNA sequence data from the pelvic flexure, the left and right ventral colon, and the left and right dorsal colon. An average of 18,330 genes were expressed across the five tissues sampled. Most of the genes showed some level of expression in all five tissues. Tissue-restricted patterns of expression were also observed. Genes with restricted expression in the left ventral and left dorsal colons have communication, signaling, and regulatory functions that correlate with their known physiology. In contrast, genes expressed exclusively in the pelvic flexure have diverse functions. The ontology of genes differentially expressed between the pelvic flexure and the surrounding tissues was associated with immune functions and signaling processes. Despite being non-significant, these enrichment trends were reinforced by the functions of statistically significant expression differences between tissues of the hindgut. These results provide insight into the physiology of the equine hindgut epithelium that might influence the microbiota and its distribution. Full article

(This article belongs to the Special Issue Advances in Equine Genetics and Breeding)

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Gene expression analysis workflow. The steps highlighted in yellow involve preparing the RNA-seq data for analysis. Steps in orange detail mapping the data to the reference genome and annotation, while the steps in blue describe the gene expression prior to functional analysis. Full article ">Figure 2
Summary of sequence data generated by sample. The left Y-axis indicates the total reads generated, and the right Y-axis indicates the percent GC content (orange line). Data are grouped by tissue along the X-axis and horse (blue, green, and purple columns; see chart legend for associations). Full article ">Figure 3
The number of genes expressed in sections of the equine hindgut by (A) sample and (B) tissue. The average across all tissues was 18,330 +/− 191 genes (red dashed line). Full article ">Figure 4
Tau index value frequency distribution across hindgut locations. Tau index values ranged from 0 to 1, with 0 indicating consistent expression across hindgut tissues and 1 indicating differential expression (i.e., tissue-restricted expression in 1 of the five hindgut areas). The frequency displays how many genes were categorized at a particular tau index value. Full article ">Figure 5
Differential expression by tissue comparison. Purple indicates the total number of differentially expressed genes, green indicates a higher abundance in the first tissue of the comparison, and red indicates a higher abundance in the second tissue. Full article ">Figure 6
Principal component analysis showing expression differences between the right and left ventral colon (RVC and LVC), the pelvic flexure (PF), and the left and right dorsal colon (LDC and RDC). Full article ">Figure 7
Top 10 biological processes enriched in the comparisons of differential expression between the right ventral colon, left ventral colon, left dorsal colon, right dorsal colon, and pelvic flexure. Full article ">

10 pages, 915 KiB

Open AccessArticle

High Concentrate Supplementation during Late Pregnancy and Lambing Reduced Mortality of Triplet-Bearing Maternal Ewes

by Emmanuelle Haslin, Sarah E. Blumer, Darren Gordon, Gavin A. Kearney, Paul R. Kenyon, Lyndon J. Kubeil, Gordon Refshauge, Jason P. Trompf and Andrew N. Thompson

Animals 2024, 14(16), 2302; https://doi.org/10.3390/ani14162302 - 8 Aug 2024

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Abstract

Low survival of triplet-bearing ewes and their lambs represents lost production and a welfare issue. The effects of feed-on-offer (FOO; low: 1205 vs. high: 1980 kg DM/ha) and concentrate supplementation (low: 50 vs. high: 300+ g/ewe/day) levels during late pregnancy and lambing on [...] Read more.

Low survival of triplet-bearing ewes and their lambs represents lost production and a welfare issue. The effects of feed-on-offer (FOO; low: 1205 vs. high: 1980 kg DM/ha) and concentrate supplementation (low: 50 vs. high: 300+ g/ewe/day) levels during late pregnancy and lambing on the survival of triplet-bearing ewes and their lambs were investigated on 10 commercial farms using 1772 triplet-bearing Maternal ewes. Ewe and lamb survival were estimated at marking, and ewe body condition score (BCS) was recorded in late pregnancy and at marking. Although FOO treatment had no effect on triplet-bearing ewe mortality, receiving higher supplementation decreased mortality by 40% and increased BCS at marking by 0.14 compared with a lower supplementation (p < 0.05). Supplementation, FOO treatments, weather conditions during lambing and shelter availability had no effect on triplet-lamb survival. These findings suggest no additional benefit to triplet-bearing ewe survival when FOO levels exceed 1200 kg DM/ha during late pregnancy and lambing, but increased supplementation can reduce ewe mortality. Further research is required to determine the response to the supplementation level at lower FOO levels on triplet-bearing Merino ewes and their lambs and establish whether supplementation of triplet-bearing ewes during late pregnancy and lambing with higher levels of concentrates would be cost-effective. Full article

(This article belongs to the Special Issue Nutrition and Reproduction Relationships in Animals)

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23 pages, 321 KiB

Open AccessArticle

Public Attitudes towards and Management Strategies for Community Cats in Urban China

by Xuan Gu, Di Wu, Zilin Zhang, Guo Peng, Anru Ni, Bo Wang, Xiufan Xiong, Yujie Liu and Li Wang

Animals 2024, 14(16), 2301; https://doi.org/10.3390/ani14162301 - 7 Aug 2024

Viewed by 418

Abstract

Managing community cats in urban China is a contentious and emerging issue, with debates centering on the most effective and humane approaches. This study aimed to investigate public attitudes towards community cats and various management strategies. A survey was conducted involving 5382 urban [...] Read more.

Managing community cats in urban China is a contentious and emerging issue, with debates centering on the most effective and humane approaches. This study aimed to investigate public attitudes towards community cats and various management strategies. A survey was conducted involving 5382 urban residents in China. Their attitudes towards the positive and negative roles of community cats in urban areas and their support for different management methods were examined, including trap-and-kill, taking no action, centralized management, and trap–neuter–return (TNR) and its variations. Results indicated that 63% of participants were willing to coexist with community cats, 71% opposed trap-and-kill, and 61% agreed or strongly agreed with the TNR method and its variations. Older residents or those with higher incomes were more likely to support coexistence with community cats. In contrast, younger or lower-income residents were more likely to support non-coexistence. Residents in first- or second-tier cities (e.g., Beijing, Hangzhou, and Jinan Cities in China) were more inclined to support trap-and-kill and less likely to support coexistence than their counterparts in fourth-tier cities (e.g., county-level cities in China). Moreover, those with lower education or incomes were more supportive of trap-and-kill and taking no action as the methods to manage community cats than those with relatively higher education or incomes. Those with higher incomes held more positive attitudes towards community cats and were more supportive of TNR and its variations than their counterparts with lower incomes. Males were more inclined to support trap-and-kill and taking no action and less inclined to support centralized management and TNR than females. The implications of the findings on TNR with adoption programs in urban China are discussed. These novel findings underscore the need for targeted educational campaigns to promote humane and effective management strategies, addressing public concerns and community cats’ welfare. The study’s insights are critical for informing policy and improving community cat management in urban China. Full article

(This article belongs to the Special Issue Free-Roaming Cats and Dogs: Ecology, Management and Inter-Species Interactions)

14 pages, 2466 KiB

Open AccessArticle

Predicting Conservation Status of Testudoformes under Climate Change Using Habitat Models

by Wenbo Liao, Shun Cao, Ying Jiang, Weijie Shao, Li Zhao and Chengzhi Yan

Animals 2024, 14(16), 2300; https://doi.org/10.3390/ani14162300 - 7 Aug 2024

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Abstract

Climate change promotes variations in distribution ranges, potentially leading to biodiversity loss and increased extinction risks for species. It is crucial to investigate these variations under future climate change scenarios for effective biodiversity conservation. Here, we studied the future distribution ranges of 268 [...] Read more.

Climate change promotes variations in distribution ranges, potentially leading to biodiversity loss and increased extinction risks for species. It is crucial to investigate these variations under future climate change scenarios for effective biodiversity conservation. Here, we studied the future distribution ranges of 268 Testudoformes species under climate change using habitat models, specifically species distribution models (SDMs), to assess their conservation status. Our results have indicated that over half of species are projected to experience declines in their potential distribution ranges under two scenarios. In particular, we found that three critically endangered species—Three-striped roofed turtle (Batagur dhongoka), Durango mud turtle (Kinosternon durangoense), and Colombian mud turtle (Kinosternon dunni)—displayed extraction of their distribution ranges and faced extinction under global climate change. Additionally, our analysis revealed that the potential distribution ranges of some species might increase under future climate scenarios. However, these findings must be interpreted with caution as they do not account for other significant factors such as biological invasions, population structure, land-use change, anthropogenic disturbances, and inter-organism interrelationships. Future studies should incorporate these factors to provide a more comprehensive assessment of extinction risks. Our findings suggest that climate change, in conjunction with habitat degradation and human activities, must be considered when assessing the extinction risks of Testudoformes. Full article

(This article belongs to the Section Herpetology)

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13 pages, 4286 KiB

Open AccessArticle

Equine Sarcocystosis in the Northern Region of the Republic of Kazakhstan

by Zhanaidar Bermukhametov, Kulyay Suleimanova, Oksana Tomaruk, Bakhit Baimenov, Pavel Shevchenko, Assylbek Batyrbekov, Zoja Mikniene, Ahmet Onur Girişgin and Raushan Rychshanova

Animals 2024, 14(16), 2299; https://doi.org/10.3390/ani14162299 - 7 Aug 2024

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Abstract

Background: A total of 396 samples were taken from the hearts, oesophagi, and diaphragms of 132 horses slaughtered at slaughterhouses in 2023 for subsequent examination. Methods: The histological method revealed pathomorphological changes in the muscle tissue. The molecular method identified the pathogen species. [...] Read more.

Background: A total of 396 samples were taken from the hearts, oesophagi, and diaphragms of 132 horses slaughtered at slaughterhouses in 2023 for subsequent examination. Methods: The histological method revealed pathomorphological changes in the muscle tissue. The molecular method identified the pathogen species. Results: Histological examination revealed thick-walled cysts with internal septa and numerous bradyzoites, and mononuclear inflammatory cells with pericyst infiltrates. Microcyst samples were amplified by polymerase chain reaction. Molecular genetic analysis allowed for the identification of 18 sarcocysts. Phylogenetic analysis of Sarcocystis isolates revealed three separate clades of Sarcocystis bertrami and two separate clades of Sarcocystis fayeri. Comparison and phylogenetic analysis revealed a very close relationship between the identified Sarcocystis species and other equine Sarcocystis DNA sequences from China and Japan. Based on the results obtained, the epizootic situation and the parasitic level of sarcocystosis contamination of horses in the northern Kazakhstan were determined. Conclusion: This is the first histological and molecular study to identify Sarcocystis spp. isolated from microscopic forms of equine sarcocysts in the northern Kazakhstan. This research will contribute to the fight against the spread of sarcocystosis in the Republic of Kazakhstan and will allow us to develop proposals for improving the mechanisms of sarcocystosis control. Full article

(This article belongs to the Special Issue Parasite Epidemiology and Molecules Identification in Wild and Domestic Animals)

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Sarcocystis in the cardiac tissue, first-generation staining with haematoxylin and eosin ×200. Full article ">Figure 2
Sarcocystis in the cardiac tissue, second-generation Van Gieson staining ×200. Full article ">Figure 3
Sarcocystis in the oesophagus, second-generation meronth staining with haematoxylin and eosin ×200. Full article ">Figure 4
Sarcocystis in the oesophagus, second-generation merontium Van-Gizon staining ×200. Full article ">Figure 5
Sarcocysts in the diaphragm, haematoxylin and eosin staining ×200. Full article ">Figure 6
Sarcocysts in the diaphragm Van Gieson staining ×200. Full article ">Figure 7
PCR results: A (SF1-SR9); B (SF1-SR4); C (SF1-SR5); D (SF1-COIRm); 1–23, amplified fragments; M, DNA marker 100–1000 bp. Full article ">Figure 8
A phylogenetic tree of selected isolates of Sarcocystis bertrami and Sarcocystis fayeri found in horses in the Kostanai region and representative sequences deposited in GenBank based on partial cox1 sequences using the neighbour-joining method is presented. GenBank inventory numbers are indicated next to taxon names. Full article ">

16 pages, 1771 KiB

Open AccessArticle

Estimation of Genetic Parameters of Early Growth Traits in Dumeng Sheep

by Ruijun Wang, Xinle Wang, Baodong Liu, Lifei Zhang, Jing Li, Dayong Chen, Yunhui Ma, Huijie He, Jie Liu, Yongbin Liu and Yanjun Zhang

Animals 2024, 14(16), 2298; https://doi.org/10.3390/ani14162298 - 7 Aug 2024

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Abstract

This study aimed to estimate the genetic and non-genetic factors that affect the nine early growth traits of Dumeng sheep, as well as to estimate the variance components and genetic parameters associated with these traits. A dataset containing detailed information on 17,896 preweaning [...] Read more.

This study aimed to estimate the genetic and non-genetic factors that affect the nine early growth traits of Dumeng sheep, as well as to estimate the variance components and genetic parameters associated with these traits. A dataset containing detailed information on 17,896 preweaning trait records of 4474 lambs was collected. In addition, 5015 postweaning trait records of 1003 lambs were documented. The effects of recipient dam age, sex, year, season, and herd on the early growth traits were assessed using the general linear model procedure of the statistical analysis system, revealing different levels of significance across different traits. To determine the most suitable model for estimating the genetic parameters, the likelihood ratio (LR) test was employed, fitting six animal models that either excluded or included maternal genetic and maternal permanent environmental effects within the average information restricted maximum likelihood (AIREML) framework using WOMBAT software (Version: 23/11/23). The model incorporating direct additive genetic effects, maternal genetic effects, and maternal permanent environment effects as random effects (model 6) provided the best fit for birth weight (BW) estimation. In contrast, the model combining direct additive genetic effects and maternal permanent environment effects as random effects (model 2) demonstrated a superior fit for estimating the genetic parameters of weaning weight (WW), average daily gain weight from birth to weaning (ADG1), and Kleiber ratio from birth to weaning (KR1). With regard to the genetic parameters of body weight at 6 months of age (6MW), average daily gain weight from weaning to 6 months (ADG2), average daily gain weight from birth to 6 months (ADG3), Kleiber ratio from weaning to 6 months (KR2), and Kleiber ratio from birth to 6 months (KR3), model 1, which incorporates only direct additive genetic effects, was identified as the optimal choice. With the optimal model, the heritability estimates ranged from 0.010 ± 0.033 for 6MW to 0.1837 ± 0.096 for KR3. The bivariate analysis method was employed to estimate the correlation between various traits using the most suitable model. The absolute values of genetic correlation coefficients among the traits spanned a range from 0.1460 to 0.9998, highlighting both weak and strong relationships among the studied traits. Specifically, the estimated genetic correlations between WW and ADG1, ADG3, KR1, and KR3 were 0.9859, 0.9953, 0.9911, and 0.9951, respectively, while the corresponding phenotypic correlations were 0.9752, 0.7836, 0.8262, and 0.5767. These findings identified that WW could serve as an effective selection criterion for enhancing early growth traits. Full article

(This article belongs to the Section Small Ruminants)

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15 pages, 7178 KiB

Open AccessArticle

Phagocytic Function and Flow Cytometric Phenotype of Asian Elephant Monocytes

by Jennifer L. Johns, Trinity R. Baumgartner, Carlos R. Sanchez and Brian P. Dolan

Animals 2024, 14(16), 2297; https://doi.org/10.3390/ani14162297 - 7 Aug 2024

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Abstract

Optimal veterinary care of managed elephant populations is vital due to the continued decline of wild populations. Appropriate health monitoring and accurate disease diagnosis include hematologic evaluation. Elephant hematology is distinctive in that elephants have high percentages of monocytes in health. Elephant monocytes [...] Read more.

Optimal veterinary care of managed elephant populations is vital due to the continued decline of wild populations. Appropriate health monitoring and accurate disease diagnosis include hematologic evaluation. Elephant hematology is distinctive in that elephants have high percentages of monocytes in health. Elephant monocytes also have unusual morphology, a feature shared with manatees and rock hyraxes. Manual white blood cell counting is used for elephant hematology, as analyzers are generally inaccurate. The aims of this study were to evaluate basic cell isolation and functional testing protocols for use in elephant monocyte research, and to test several available antibodies via flow cytometry for use in elephant monocyte identification. Peripheral blood samples from five Asian elephants (Elephas maximus) were used. Methods for monocyte isolation and evaluation of phagocytic function were established. Putative lymphocyte and monocyte populations were identified using a scatter on flow cytometry. Antibodies against CD11b, CD11c, CD14, and ionized calcium-binding adapter molecule 1 (IBA1) were tested, with IBA1 showing the highest apparent diagnostic utility in labeling monocytes. Combined flow cytometric scatter and IBA1 positivity appear to identify Asian elephant monocytes. These data provide a methodologic basis for further investigation into elephant monocyte function and immune response to infection. Full article

(This article belongs to the Special Issue Wildlife Clinical Pathology: A One Health Key to Ecosystem Assessment)

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Leukocytes in peripheral blood smears from Asian elephants. Wright-Giemsa stain, 100× magnification. (a–d) Monocytes; (e) Lymphocyte; (f) Heterophil (neutrophil); (g) Eosinophil; (h) Basophil. Full article ">Figure 1 Cont.
Leukocytes in peripheral blood smears from Asian elephants. Wright-Giemsa stain, 100× magnification. (a–d) Monocytes; (e) Lymphocyte; (f) Heterophil (neutrophil); (g) Eosinophil; (h) Basophil. Full article ">Figure 2
Representative cytocentrifuge smear confirming high PBMC purity following density gradient centrifugation. Modified Wright stain, 100× magnification. Arrow = heterophil. Full article ">Figure 3
Fluorescence spectrophotometry of elephant monocytes exposed to pHrodo-labeled zymosan particles. Values are normalized to “particle only” negative control wells. Monocytes from three Asian elephants (female 1 (F1), female 2 (F2), and male 1 (M1) were tested. Data points represent mean values; bars represent standard deviation. Full article ">Figure 4
Fluorescence microscopy images of elephant monocytes incubated with zymosan-coated fluorescent particles. (a–c) DAPI nuclear stain (UV excitation); (d–f) particle fluorescence (560 nm excitation); (g–i) merged images of above two rows; (j–l) contrast images; (m–o) merged images of all above images. Full article ">Figure 4 Cont.
Fluorescence microscopy images of elephant monocytes incubated with zymosan-coated fluorescent particles. (a–c) DAPI nuclear stain (UV excitation); (d–f) particle fluorescence (560 nm excitation); (g–i) merged images of above two rows; (j–l) contrast images; (m–o) merged images of all above images. Full article ">Figure 5
Flow cytometric characterization of Asian elephant peripheral blood mononuclear cells (PBMCs). (a) Singlet events gated by forward scatter area (x-axis) versus forward scatter height (y-axis); (b) gating for viability (blue peak = representative sample with viability dye, red peak = sample without viability dye); (c) PBMC subsets defined via forward scatter area (x-axis) and side scatter area (y-axis). Full article ">Figure 6
Phagocytosis of zymosan-coated fluorescent particles in lymphocyte and monocyte populations gated via scatter. (a) Elephant F1; (b) Elephant F2; (c) Elephant M1. Red peak = lymphocytes; blue peak = monocytes; “FITC+” = percentage of positive events for particle fluorescence in monocyte population. Full article ">Figure 7
Positive events for surface phenotype markers in lymphocyte and monocyte populations gated via scatter. (a) IBA1 positivity, elephant F1; (b) IBA1 positivity, elephant F2; (c) IBA1 positivity, elephant M1; (d) representative histogram for CD11b; (e) representative histogram for CD11c; (f) representative histogram for CD14. Red peak = lymphocytes; blue peak = monocytes. Full article ">

19 pages, 1962 KiB

Open AccessArticle

Effect of Different Slow-Release Urea on the Production Performance, Rumen Fermentation, and Blood Parameter of Angus Heifer

by Caiyun Fan, Hongguang Li, Shuaihong Li, Gang Zhong, Wenbin Jia, Zhao Zhuo, Yanfeng Xue, Anne F. Koontz and Jianbo Cheng

Animals 2024, 14(16), 2296; https://doi.org/10.3390/ani14162296 - 7 Aug 2024

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Abstract

This study investigated the effect of replacing part of the dietary soybean meal with either polymer-coated urea or gelatinized starch urea on the production performance, blood indexes, and ruminal fermentation of Angus heifers. A total of 210 purebred Angus cattle (BW = 314.26 [...] Read more.

This study investigated the effect of replacing part of the dietary soybean meal with either polymer-coated urea or gelatinized starch urea on the production performance, blood indexes, and ruminal fermentation of Angus heifers. A total of 210 purebred Angus cattle (BW = 314.26 kg) were divided into three groups: the no urea group (CON), the polymer-coated urea group (PCU), and the gelatinized starch urea group (GSU); 20 g/kg polymer-coated urea or 25 g/kg gelatinized starch urea was used to replace part of soybean meal in the concentrate feed, according to the principle of isometabolic energy and isonitrogenous. The result showed that the PCU group had higher ADG and ADF apparent digestibility, while it had a lower feed–weight ratio. On the 86th day, the serum albumin (ALB) content in the PCU group was significantly higher than that in the CON group. In rumen, compared with the CON group, the contents of acetic acid and total volatile fatty acid were significantly higher in the PCU group, whereas butyric acid and propionic acid were significantly higher in the PCU group and GSU group. Ruminal bacterial diversity analysis found that the abundance of Firmicutes was higher in the PCU group at the phylum level, and an inverse result was observed in Bacteroidetes. The abundance of Paraprevotella was higher in the PCU group, whereas higher abundance of Prevotella was found in the GSU group at the genus level. These results indicate that slow-release urea can replace part of soybean meal in the diet, and the amount of substitution in this trial had no diverse effect on the performance of Angus heifers. Full article

(This article belongs to the Section Cattle)

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20 pages, 2399 KiB

Open AccessArticle

The Use of Machine Learning to Predict Prevalence of Subclinical Mastitis in Dairy Sheep Farms

by Yiannis Kiouvrekis, Natalia G. C. Vasileiou, Eleni I. Katsarou, Daphne T. Lianou, Charalambia K. Michael, Sotiris Zikas, Angeliki I. Katsafadou, Maria V. Bourganou, Dimitra V. Liagka, Dimitris C. Chatzopoulos and George C. Fthenakis

Animals 2024, 14(16), 2295; https://doi.org/10.3390/ani14162295 - 6 Aug 2024

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Abstract

The objective of the study was to develop a computational model with which predictions regarding the level of prevalence of mastitis in dairy sheep farms could be performed. Data for the construction of the model were obtained from a large Greece-wide field study [...] Read more.

The objective of the study was to develop a computational model with which predictions regarding the level of prevalence of mastitis in dairy sheep farms could be performed. Data for the construction of the model were obtained from a large Greece-wide field study with 111 farms. Unsupervised learning methodology was applied for clustering data into two clusters based on 18 variables (17 independent variables related to health management practices applied in farms, climatological data at the locations of the farms, and the level of prevalence of subclinical mastitis as the target value). The K-means tool showed the highest significance for the classification of farms into two clusters for the construction of the computational model: median (interquartile range) prevalence of subclinical mastitis among farms was 20.0% (interquartile range: 15.8%) and 30.0% (16.0%) (p = 0.002). Supervised learning tools were subsequently used to predict the level of prevalence of the infection: decision trees, k-NN, neural networks, and Support vector machines. For each of these, combinations of hyperparameters were employed; 83 models were produced, and 4150 assessments were made in total. A computational model obtained by means of Support vector machines (kernel: ‘linear’, regularization parameter C = 3) was selected. Thereafter, the model was assessed through the results of the prevalence of subclinical mastitis in 373 records from sheep flocks unrelated to the ones employed for the selection of the model; the model was used for evaluation of the correct classification of the data in each of 373 sets, each of which included a test (prediction) subset with one record that referred to the farm under assessment. The median prevalence of the infection in farms classified by the model in each of the two categories was 10.4% (5.5%) and 36.3% (9.7%) (p < 0.0001). The overall accuracy of the model for the results presented by the K-means tool was 94.1%; for the estimation of the level of prevalence (<25.0%/≥25.0%) in the farms, it was 96.3%. The findings of this study indicate that machine learning algorithms can be usefully employed in predicting the level of subclinical mastitis in dairy sheep farms. This can facilitate setting up appropriate health management measures for interventions in the farms. Full article

(This article belongs to the Special Issue Advances in Mammary Gland Biology and Lactation of Ruminants: Second Edition)

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Location of 111 sheep farms around Greece that were included in a countrywide investigation on subclinical mastitis, records from which were used in the construction of the computational model. Full article ">Figure 2
Box and whisker plots of the two clusters of sheep farms according to level of prevalence of subclinical mastitis, created by applying unsupervised learning tools (from left to right: Affinity propagation (blue), Birch threshold 3 (purple), Birch threshold 4 (yellow), Hierarchical clustering (orange), K-means (azure), Spectral clustering (red), and Spectral clustering rbf (green)). Full article ">Figure 3
Location of 325 sheep farms around Greece that were included in a countrywide investigation, records from which were used in the assessment (verification) of the computational model selected. Full article ">Figure 4
Box and whisker plots of the data for measures of quality in each of the four models selected after evaluation within each tool used during assessment for classification of records from sheep farms based on prevalence of subclinical mastitis (green: decision trees, purple: k-NN, blue: neural networks, pink: Support vector machines—full pattern: accuracy, dotted pattern: precision, striped pattern: recall). Full article ">Figure 5
Box and whisker plots of the two categories of sheep farms according to predicted level (i.e., low/high) of prevalence of subclinical mastitis, created by using Support vector machines (green: farms assigned in category with low level of prevalence; pink: farms assigned in category with high level of prevalence). Full article ">Figure 6
SHapley Additive exPlanations values for the importance of each of 17 independent variables in the prediction of the level (i.e., low/high) of prevalence of subclinical mastitis (B: breed of ewes, G: vaccination against staphylococcal mastitis, C: management system applied in farm, M: administration of antibiotics at the end of the lactation period, L: application of measures for mastitis control at the end of the lactation period, E: application of reproductive control, Y: altitude of farm location, X: wind speed at farm location, D: month of lactation period, V: annual precipitation at farm location, A: no. of ewes in farm, J: milking status of the ewes in farm, K: application of teat dipping; not included in output: minimum temperature of coldest month at farm location, distance of farm from other sheep farms, land use at farm location, and microhabitat at farm location) (class 0/1: low (<25.0%)/high (≥25.0%) level of prevalence of subclinical mastitis) during assessment (verification) of a computational machine learning model. Full article ">

13 pages, 1663 KiB

Open AccessArticle

Study of Plasma Biochemistry and Plasma Metabolomics Differences in Montbéliard and Holstein Backcross and Holstein Heifers

by Haihui Wang, Haomiao Chang, Hantong Weng, Yunfei Zhai, Hanfang Zeng, Shujie Li and Zhaoyu Han

Animals 2024, 14(16), 2294; https://doi.org/10.3390/ani14162294 - 6 Aug 2024

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Abstract

Holstein cattle are the main breed of dairy cattle in China. However, given the high degree of purebred selection of Holstein cattle, Chinese dairy cattle are increasingly being characterized by poor disease resistance, poor quality, and declining fertility. In this study, using Montbéliard [...] Read more.

Holstein cattle are the main breed of dairy cattle in China. However, given the high degree of purebred selection of Holstein cattle, Chinese dairy cattle are increasingly being characterized by poor disease resistance, poor quality, and declining fertility. In this study, using Montbéliard × Holstein cattle as females and Montbéliard bulls as males for backcross breeding, we sought to provide a reference for improving the quality and performance of Holstein cattle and enhancing the efficiency of dairy farming. On the basis of similar physiological status and age, we selected 24 Montbéliard and Holstein backcross heifers and 11 Holstein heifers fed the same formula for comparative analyses. Plasma samples collected for plasma biochemical index analyses revealed that the content of ALB and BUN in the Montbéliard and Holstein backcross heifers was 20.83% (31.62 g/L to 26.17 g/L) and 42.36% (6.89 mmol/L to 4.84 mmol/L) higher than in the Holsteins (p < 0.01). The ALB/GLB (0.90 to 0.60, p < 0.05) was significantly higher in Montbéliard and Holstein backcross heifers than in Holstein heifers. Similarly, the activity of CAT in the backcross heifers was 61.28% (4.29 U/mL to 2.66 U/mL) higher than that in the Holstein heifers (p < 0.05). Although the activity of GSH-Px in the backcross heifers also showed an increasing trend, the difference did not reach the level of statistical significance (p = 0.052). Compared with Holstein heifers, the concentrations of IgA, IgG, and IL-4 were elevated by 32.52% (24.90 μg/mL to 18.79 μg/mL, p < 0.01), 13.46% (234.32 μg/mL to 206.53 μg/mL, p < 0.01), and 14.59% (306.27 pg/mL to 267.28 pg/mL, p < 0.05), and the contents of IL-6 and TNF-α were decreased by 15.92% (215.71 pg/mL to 256.55 pg/mL, p < 0.01) and 32.17% (7.17 ng/mL to 10.57 ng/mL, p < 0.01) in the plasma of Montbéliard and Holstein backcross heifers. Among the experimental heifers, five animals from each of the two groups were selected for plasma metabolomic analysis based on untargeted liquid chromatography–mass spectrometry. A comparison of the differential metabolites between the two heifer breeds revealed an up-regulation of d-glucuronic acid, s-glutathionyl-l-cysteine, and oleic acid levels in the backcross cattle compared with those in the Holstein heifers. We speculate that changes in the levels of these metabolites may be associated with an enhancement of the anti-inflammatory, antioxidant, and immune systems in these backcross heifers. Collectively, our findings in this study indicate that compared with 12-month-old purebred Holstein heifers, Montbéliard and Holstein backcross heifers of the same age are characterized by higher antioxidant capacity and immunity. Full article

(This article belongs to the Section Cattle)

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33 pages, 6399 KiB

Open AccessArticle

Effects of Early Lactation Milking Frequency in an Automated Milking System on Cow Performance

by Erin M. Kammann, Elizabeth A. French, Natalie S. Jozik, Wenli Li and Ryan S. Pralle

Animals 2024, 14(16), 2293; https://doi.org/10.3390/ani14162293 - 6 Aug 2024

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Abstract

Automated milking systems (AMS) are increasingly adopted for dairy cow production, promoting individualized cow management dependent on factors like lactation stage, age, and productivity. The study objective was to investigate the effects of early lactation milking frequency on cows milked via [...] Read more.

Automated milking systems (AMS) are increasingly adopted for dairy cow production, promoting individualized cow management dependent on factors like lactation stage, age, and productivity. The study objective was to investigate the effects of early lactation milking frequency on cows milked via AMS. Multiparous Holstein cows blocked by parity and due date were randomly assigned to treatments (n = 8 per treatment): three (3X) or six (6X) milkings per day (MPD). The experimental phase (EXP) was defined as 4 to 29 days in milk (DIM). The AMS settings were programed so 3X cows were limited to three MPD while 6X cows were allowed six MPD. Afterwards was the carry over phase (CO) ranging from 30 to 90 DIM; all cows were allowed up to six MPD. Measurements by the AMS included bodyweight, milk yield (MY), and pellet intake. Weekly composite milk samples were analyzed for macronutrient composition and fatty acid (FA) profile. Coccygeal blood was sampled at 3, 8 ± 1, and 13 ± 1 DIM; concentrations of blood plasma analytes were quantified. Greater MPD was achieved for 6X cows versus 3X cows during EXP, but similar during the CO. Daily MY was non-separable during the EXP while 6X cows in their third or greater lactation group (3 + LG) had greater MY than 3X cows of the same LG during the CO. Milk fat content and 4% fat-corrected MY were both greater for 6X, 3 + LG cows during the EXP compared to 3X, 3 + LG cows. Milk FA methyl esters (FAME) proportions were different between MPD groups, with 6X, 3 + LG cows having the lowest short, even-chain FA from de novo or post-absorptive origin. Differences in analytes indicated that 6X, 3 + LG cows experienced metabolic stress and incorporated greater FA from adipose tissue. Greater early lactation MPD in AMS may shift cow nutrient partitioning to support greater production in 3+ parity cows. Full article

(This article belongs to the Special Issue Improving Sustainability of Ruminant Agriculture: New Strategies and Tools)

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12 pages, 270 KiB

Open AccessReview

Degnala in Water Buffaloes: A Review on a Neglected Disease

by André de Medeiros Costa Lins and Felipe Masiero Salvarani

Animals 2024, 14(16), 2292; https://doi.org/10.3390/ani14162292 - 6 Aug 2024

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Abstract

Degnala is one of the primary mycotoxicoses affecting buffaloes, with Fusarium spp. as the main causative agent. This disease is strongly associated with the feeding of rice straw to buffaloes and is considered endemic to regions where rice is cultivated. Cases are concentrated [...] Read more.

Degnala is one of the primary mycotoxicoses affecting buffaloes, with Fusarium spp. as the main causative agent. This disease is strongly associated with the feeding of rice straw to buffaloes and is considered endemic to regions where rice is cultivated. Cases are concentrated in winter when conditions favor fungal growth in inadequately stored straw. Degnala is characterized by necrosis and gangrene of the extremities, including the tail, lower limbs, ears, tongue, muzzle, and teats. The pelvic limbs are more affected than the thoracic limbs. A tortuous appearance of the tail is very common, and cracks or corneal loss of the hooves may occur, exposing the blades and even the bones. There is no diagnostic method for identifying the disease in animals other than clinical and epidemiological criteria, combined with fungal culture samples. There is no treatment that neutralizes the effects of the toxins; the current treatment is palliative and supportive, consisting of pentasulfate solution, anti-Degnala liquid, 2% nitroglycerin ointment, and broad-spectrum antibiotics for secondary infections. Additionally, the management of drying and proper storage of straw is essential for controlling this disease. Full article

(This article belongs to the Special Issue Water Buffalo Welfare, Strategies to Improve Health, Behavior, Productivity, and Food Quality and Safety)

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Animals, Volume 14, Issue 16 (August-2 2024) – 39 articles

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