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Animals, Volume 10, Issue 8 (August 2020) – 201 articles

Cover Story (view full-size image): Aggression between farmed pigs is common during regrouping but rarely severe in stable social groups. We report the occurrence of lethal gang aggression in groups of familiar pigs, whereby the group fatally attacks one member. We document this extreme aggression and suggest potential causes by farmer surveys and information on victims’ injuries and body condition. Gang aggression occurred more on farms with deep straw bedding; however, this may also be related to other factors, which could not be discerned. Gang aggression was not limited to specific genetics, breed, or feed type. It equally occurred between females and males and tended to occur more in winter. Dead victims were covered in injuries, but had a healthy body condition, whereas survivors had a lower body condition score. We hope this study will stimulate work to understand lethal gang aggression. Photo by SRUC. View this paper
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17 pages, 3274 KiB  
Article
B-Mode and Contrast Enhanced Ultrasonography Features of Gastric Inflammatory and Neoplastic Diseases in Cats
by Francesco Simeoni, Rossella Terragni, Giuseppe Rubini, Roberto Tamburro, Francesca Del Signore, Ilaria Falerno, Giovanni Aste, Marco Russo, Giovanni Mastromatteo and Massimo Vignoli
Animals 2020, 10(8), 1444; https://doi.org/10.3390/ani10081444 - 18 Aug 2020
Cited by 7 | Viewed by 4634
Abstract
Alimentary lymphoma (AL) is the most common malignancy of the feline gastrointestinal tract and may cause variable mild to severe alteration of the gastric wall on ultrasonography (US) that can be very similar to those caused by inflammation (INF). The aim of this [...] Read more.
Alimentary lymphoma (AL) is the most common malignancy of the feline gastrointestinal tract and may cause variable mild to severe alteration of the gastric wall on ultrasonography (US) that can be very similar to those caused by inflammation (INF). The aim of this prospective study is to establish the value of B-mode and contrast-enhanced US (CEUS) in describing specific features of normal, inflammatory, and neoplastic gastric diseases in feline species. B-mode US and CEUS of the stomach were performed in anesthetized cats with or without gastric disorders. Gastric wall qualitative and quantitative parameters were evaluated on B-mode US and CEUS examination. A total of 29 cats were included: six healthy (HEA) cats as the control group; nine INF; three low-grade lymphoma (LGAL); 10 high-grade lymphoma (HGAL). On B-mode US, there were significant differences in thickness, the wall’s layer definition and echogenicity between HGAL and all the other groups (<0.001). For CEUS, statistical differences between groups were found in the following: HGAL vs. HEA, HGAL vs. INF; HGAL vs. LGAL; INF vs. HEA. Diagnostic accuracy (AUC) and cut-off value were calculated and found to be significant for thickness (3.8 mm) for INF vs. LGAL (AUC > 0.70) and “benign” vs. “malignant” (AUC > 0.90) as well as peak enhancement (34.87 dB) for “benign” vs. “malignant” (AUC > 0.70). INF and LGAL showed an overlap of qualitative and quantitative parameters both on B-mode and CEUS, while HGAL usually appears as a severe wall thickening with absent layer definition, high-contrast uptake, a specific enhancement pattern, regional lymphadenopathy and local steatitis. Thickness and peak enhancement can be useful parameters in the characterization of gastric infiltrates in cats. Full article
(This article belongs to the Special Issue Oncology in Veterinary Medicine)
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<p>Examples of time–intensity curve explaining how peak enhancement (PE), wash-in rate (WiR), arrival time (AT), rising time (RT), time to peak (TTP), falling time (FT) and time to fall (TO) were calculated.</p>
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<p>Cat affected by gastric low-grade lymphoma (LGAL) showing: (<b>A</b>) diffuse reduced layer definition associated with mild increases in wall thickness (3.8 mm between the asterisks) on B-mode examination; (<b>B</b>,<b>C</b>) B-mode (<b>B</b>) and contrast-enhanced ultrasonography (CEUS) (<b>C</b>) clip acquired using dual-mode visualization in order to better identify the gastric wall during CEUS examination.</p>
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<p>Feline gastric high-grade lymphoma (HGAL): (<b>A</b>) B-mode image of a severe and diffuse hypoechoic wall thickening of 18 mm (between the asterisks) with complete loss of normal wall layers; (<b>B</b>) CEUS examination of the same cat during the peak enhancement of time–intensity curve.</p>
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<p>Box and whisker plot of the gastric wall thickness for healthy (HEA), inflammatory (INF), low-grade alimentary lymphoma (LGAL) and high-grade alimentary lymphoma (HGAL). <sup>a,b,c</sup> Values with different superscript letters differ significantly (<span class="html-italic">p</span> &lt; 0.001).</p>
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<p>Classification of gastric wall layer definition on B-mode US: (<b>A</b>) normal: all the five layers are easily identified and thickness is normal (2.4 mm between the cursors in a HEA cat); (<b>B</b>) reduced: the identification of the layers is more difficult and, in some portions, it may not be evident, in addition to a mild diffuse wall thickening (3.6 mm between the cursors in a INF cat); (<b>C</b>) absent: it is not possible to recognize the normal wall stratification and layer definition is lost. Here, a focal transmural mass (16.5 mm between the cursors in a HGAL cat) with mixed echogenicity is also present.</p>
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<p>Different localization in gastric feline HGAL: (<b>A</b>) focal: a 7-year-old cat with a transmural mass (11.4 mm) with mixed echogenicity and loss of wall layers; (<b>B</b>) diffuse: severe hypoechoic thickening (20.8 mm) of gastric body with complete loss of normal layers in a cat 12-year-old cat.</p>
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<p>Box and whisker plot of the falling time (FT) for healthy (HEA), inflammatory (INF), low-grade alimentary lymphoma (LGAL) and high-grade alimentary lymphoma (HGAL). <sup>a,b</sup> Values with different superscript letters differ significantly (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Box and whisker plot of the time to fall (TO) for healthy (HEA), inflammatory (INF), low-grade alimentary lymphoma (LGAL) and high-grade alimentary lymphoma (HGAL). <sup>a,b</sup> Values with different superscript letters differ significantly (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Box and whisker plot of the peak enhancement (PE) for healthy (HEA), inflammatory (INF), low-grade alimentary lymphoma (LGAL) and high-grade alimentary lymphoma (HGAL). <sup>a,b,c,d</sup> Values with different superscript letters differ significantly (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Box and whisker plot of the wash-in rate (WiR) for healthy (HEA), inflammatory (INF), low-grade alimentary lymphoma (LGAL) and high-grade alimentary lymphoma (HGAL). <sup>a,b,c,d</sup> Values with different superscript letters differ significantly (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>CEUS enhancement pattern in a 7-year-old cat with gastric HGAL: (<b>A</b>) before contrast medium (CM) injection (0 s), the gastric mass (14.8 mm between arrowheads) and the adjacent liver parenchyma (asterisk) are visualized; (<b>B</b>) after CM injection (8 s), uptake occurs first in the liver and the gastric mass is classified as hypo-enhancing; (<b>C</b>) when CM arrives at the lesion site (14 s), comb teeth-like vessels are present (arrows); (<b>D</b>) at peak enhancement time (18 s), arterial branching is not visible anymore and enhancement homogeneity is classified as heterogeneous.</p>
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33 pages, 2730 KiB  
Article
Large Lemurs: Ecological, Demographic and Environmental Risk Factors for Weight Gain in Captivity
by Emma L. Mellor, Innes C. Cuthill, Christoph Schwitzer, Georgia J. Mason and Michael Mendl
Animals 2020, 10(8), 1443; https://doi.org/10.3390/ani10081443 - 18 Aug 2020
Cited by 7 | Viewed by 5696
Abstract
Excessive body mass, i.e., being overweight or obese, is a health concern associated with issues such as reduced fertility and lifespan. Some lemur species are prone to extreme weight gain in captivity, yet others are not. To better understand species- and individual-level effects [...] Read more.
Excessive body mass, i.e., being overweight or obese, is a health concern associated with issues such as reduced fertility and lifespan. Some lemur species are prone to extreme weight gain in captivity, yet others are not. To better understand species- and individual-level effects on susceptibility to captive weight gain, we use two complementary methods: phylogenetic comparative methods to examine ecological explanations for susceptibility to weight gain across species, and epidemiological approaches to examine demographic and environment effects within species. Data on body masses and living conditions were collected using a survey, yielding useable data on 675 lemurs representing 13 species from 96 collections worldwide. Data on species-typical wild ecology for comparative analyses came from published literature and climate databases. We uncovered one potential ecological risk factor: species adapted to greater wild food resource unpredictability tended to be more prone to weight gain. Our epidemiological analyses on the four best-sampled species revealed four demographic and one environmental risk factors, e.g., for males, being housed with only fixed climbing structures. We make practical recommendations to help address weight concerns, and describe future research including ways to validate the proxy we used to infer body condition. Full article
(This article belongs to the Special Issue Evidence-Based Practice in Zoo Animal Management)
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<p>Images of different adult lemurs in one of three body conditions as deemed by their relative body masses: healthy, overweight, or obese. The number at the bottom of each image is that animal’s relative body mass at the time the photograph was taken. Sex of lemurs (credits). <b>Far left</b>, <b>top</b>: female (Lakeland Wildlife Oasis); <b>middle</b>: male (Nancy Nill, Palm Beach Zoo); <b>bottom</b>: female (Ashley Ashcraft). <b>Middle</b>, <b>top</b>: male (Valerie Schultz, Smithsonian’s National Zoo); <b>middle</b>: female (Śląski Ogród Zoologiczny); <b>bottom</b>: male (Heidi Beal). <b>Right</b>, <b>top</b>: male (Nancy Nill, Palm Beach Zoo); <b>second from top</b>: female (Śląski Ogród Zoologiczny); <b>second from bottom</b>: male (Debbie Fenton); <b>bottom</b>: male (Rebecca Lambert, Taronga Conservation Society).</p>
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<p>Box-and-whisker plots of relative body mass values of individual adult lemurs, from the 13 species featuring in our comparative analyses. Ring-tailed lemurs, mongoose lemurs, black-and-white ruffed lemurs, and red ruffed lemurs also featured in our epidemiological analyses. Sample sizes are shown in parenthesis next to species’ names, and are also are indicated by the relative width of their respective boxplot. Species’ medians are indicated by vertical lines on the boxes; the extent of the boxes indicate their interquartile ranges; whiskers represent values within 1.5 times the interquartile range; and outliers outside this are depicted as points. The healthy range (0.75–1.25) is shown by yellow shading; overweight by orange (1.25–1.5); obese by red (1.5–2); and morbidly obese (&gt;2) by the darker red.</p>
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<p>Phylogenetic tree of 13 lemur species in our comparative analyses, with their species-typical median relative body mass values shown as tip-points. The colour of a species’ tip-point represents its species-typical body condition: the healthy range (species-typical relative body mass 0.75–1.25) is shown by yellow shading; overweight by orange (1.25–1.5); obese by red (1.5–2); and morbidly obese (&gt;2) by the darker red.</p>
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<p>Relationship between between-year rainfall coefficient of variation and species-typical median relative body mass across 13 lemur species (<span class="html-italic">t</span><sub>11</sub> = 2.04, R<sup>2</sup> = 0.27, λ &lt; 0.001, <span class="html-italic">p</span> = 0.07). The shaded area shows the 95% confidence region. <span class="html-italic">Labels</span>: AGL: Alaotran gentle lemur; AA: aye aye; BEBL: blue-eyed black lemur; BL: black lemur; BrL: brown lemur; BWRL: black-and-white ruffed lemur; CL: crowned lemur; RCL: red-collared lemur; RCL: Coquerel’s sifaka; ML: mongoose lemur; RBL: red-bellied lemur; RRL: red ruffed lemur; RTL: ring-tailed lemur.</p>
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<p>Relationships between relative body mass and species × sex, age, climbing structure provision × sex, and season × sex. Dotted horizontal line indicates the point at which relative body mass = 1 (i.e., the captive animal is the same weight as its species-typical wild mean; 0 on the log-scale); the short dashed line indicates the overweight threshold (≥1.25, or 0.22 on the log-scale); and the long dashed line indicates the obese threshold (≥1.5, or 0.41 on the log-scale). On the three plots with error bars, points indicate the mean relative body mass value, and the whiskers the upper and lower 95% confidence intervals; differences between groups are indicated with asterisks; and females are indicated by black, males by grey. <b>Top Left</b>: BWRL: black-and-white ruffed lemur. ML: mongoose lemur. RRL: red ruffed lemur. RTL: ring-tailed lemur. Male mongoose lemurs have smaller relative body masses than females (<span class="html-italic">t</span><sub>6</sub> = −2.76, <span class="html-italic">p</span> = 0.03), whereas male ring-tailed lemurs have larger values than females (<span class="html-italic">t</span><sub>127</sub> = 3.15, <span class="html-italic">p</span> &lt; 0.01). <b>Top Right</b>: Relative body mass increases with age (<span class="html-italic">t</span><sub>157</sub> = 2.09, <span class="html-italic">p</span> = 0.04) across four lemur species (both log-transformed): black-and-white ruffed lemur = filled square; mongoose lemur = filled triangle; red ruffed lemur = filled circle; ring-tailed lemur = unfilled circle. The shaded area shows the 95% confidence region. <b>Bottom Left</b>: Males housed in enclosures featuring fixed climbing structures only (versus flexible and fixed structures) have larger relative body mass values (<span class="html-italic">t</span><sub>157</sub> = 3.04, <span class="html-italic">p</span> &lt; 0.01). <b>Bottom Right</b>: Females weighed in the winter had larger relative body masses than those weighed in spring (<span class="html-italic">t</span><sub>178</sub> = 3.30, <span class="html-italic">p</span> &lt; 0.01) and summer (<span class="html-italic">t</span><sub>25</sub> = 3.70, <span class="html-italic">p</span> &lt; 0.01).</p>
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12 pages, 712 KiB  
Article
Stress Associated with Simulated Transport, Changes Serum Biochemistry, Postmortem Muscle Metabolism, and Meat Quality of Broilers
by Aijuan Zheng, Shumei Lin, Shoaib Ahmed Pirzado, Zhimin Chen, Wenhuan Chang, Huiyi Cai and Guohua Liu
Animals 2020, 10(8), 1442; https://doi.org/10.3390/ani10081442 - 18 Aug 2020
Cited by 17 | Viewed by 4944
Abstract
Many factors contribute to the stress of transporting broilers from the farm to the processing plant. Using a motion simulation machine, a total of 144 male broilers were employed to determine the effect of motion, vibration, and feed withdrawal during transportation on serum [...] Read more.
Many factors contribute to the stress of transporting broilers from the farm to the processing plant. Using a motion simulation machine, a total of 144 male broilers were employed to determine the effect of motion, vibration, and feed withdrawal during transportation on serum biochemical parameters, postmortem muscle metabolism, and meat quality of broilers. The results indicated that transportation did not affect the activity of lactate dehydrogenase, γ-glutamyl transferase, aspartate aminotransferase, creatine kinase, and glucose in the serum, glutathione peroxidase in the breast and thigh muscle, nitric oxide synthase (NOS) in the breast, and heat stress protein 70 mRNA expression level in the liver (p > 0.05). Serum triiodothyronine, thyroxine, and insulin concentration declined with 2 h transportation (p < 0.05) and recovered with 4 h transportation (p < 0.05). NOS concentration in the thigh increased with 2 h transportation (p < 0.05) and recovered with 4 h transportation (p < 0.05). Two-hour and 4 h transportation increased the activity of superoxide dismutase in both muscles. Malondialdehyde, lactic acid, and drip loss24 h in both thigh and breast muscles increased, and glycogen in both muscles decreased with increasing transportation times (p < 0.05). Two-hour transportation did not influence pH45 min and pH24 h in the breast and thigh muscle, but these indexes decreased with 4 h transportation. This experiment supports and extends previous work that identified transportation as a major risk in relation to bird welfare and meat quality. Full article
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<p>Effect of different transportation treatments on the blood hormones and metabolites of broilers. (<b>A</b>) changes of T<sub>3</sub>, T<sub>4</sub>, INS, Glucose, and GGT concentration in the serum and (<b>B</b>) changes of LDH, AST, and CK concentration in the serum. Bars within a cluster with different superscripts differ (<span class="html-italic">p</span> &lt; 0.05). T<sub>3</sub> = triiodothyronine, T<sub>4</sub> = thyroxine, INS = insulin, GGT = γ-glutamyl transferase; LDH = lactate dehydrogenase, AST = asparate aminotransferase, and CK = creatine kinase.</p>
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<p>Effect of different transportation treatments on activities of GSH and SOD enzymes from breast and thigh muscle of broilers. Bars within a cluster with different superscripts differ (<span class="html-italic">p</span> &lt; 0.05). GSH = glutathione peroxidase, and SOD = superoxide dismutase.</p>
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16 pages, 4110 KiB  
Article
Genome-Wide SNP Analysis Reveals the Population Structure and the Conservation Status of 23 Italian Chicken Breeds
by Filippo Cendron, Francesco Perini, Salvatore Mastrangelo, Marco Tolone, Andrea Criscione, Salvatore Bordonaro, Nicolaia Iaffaldano, Cesare Castellini, Margherita Marzoni, Arianna Buccioni, Dominga Soglia, Achille Schiavone, Silvia Cerolini, Emiliano Lasagna and Martino Cassandro
Animals 2020, 10(8), 1441; https://doi.org/10.3390/ani10081441 - 18 Aug 2020
Cited by 33 | Viewed by 5618
Abstract
The genomic variability of local Italian chicken breeds, which were monitored under a conservation plan, was studied using single nucleotide polymorphisms (SNPs) to understand their genetic diversity and population structure. A total of 582 samples from 23 local breeds and four commercial stocks [...] Read more.
The genomic variability of local Italian chicken breeds, which were monitored under a conservation plan, was studied using single nucleotide polymorphisms (SNPs) to understand their genetic diversity and population structure. A total of 582 samples from 23 local breeds and four commercial stocks were genotyped using the Affymetrix 600 K Chicken SNP Array. In general, the levels of genetic diversity, investigated through different approaches, were lowest in the local chicken breeds compared to those in the commercial stocks. The level of genomic inbreeding, based on runs of homozygosity (FROH), was markedly different among the breeds and ranged from 0.121 (Valdarnese) to 0.607 (Siciliana). In all breeds, short runs of homozygosity (ROH) (<4 Mb in length) were more frequent than long segments. The patterns of genetic differentiation, model-based clustering, and neighbor networks showed that most breeds formed non-overlapping clusters and were clearly separate populations, which indicated the presence of gene flow, especially among breeds that originated from the same geographical area. Four genomic regions were identified as hotspots of autozygosity (islands) among the breeds, where the candidate genes are involved in morphological traits, such as body weight and feed conversion ratio. We conclude that the investigated breeds have conserved authentic genetic patterns, and these results can improve conservation strategies; moreover, the conservation of local breeds may play an important role in the local economy as a source of high-quality products for consumers. Full article
(This article belongs to the Special Issue Characterization and Strategies to Preserve Local Poultry Breeds)
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<p>Schematic representation of chicken breed origins in Italy (figure taken from <a href="http://www.d-maps.com" target="_blank">http://www.d-maps.com</a> and adapted for illustrative purposes only). For a full definition of breeds, see <a href="#animals-10-01441-t001" class="html-table">Table 1</a>.</p>
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<p>Genetic relationships among the 27 chicken breeds in this study as inferred by multidimensional scaling (MDS) analysis using (<b>A</b>) the breed-average coordinates of eigenvalues of C1 and C2 and (<b>B</b>) all of the individuals per breed. Breed acronyms are reported in <a href="#animals-10-01441-t001" class="html-table">Table 1</a>.</p>
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<p>Maximum likelihood estimation calculated with the admixture algorithm. The inferred clusters (<span class="html-italic">K</span>) were represented from <span class="html-italic">K</span> = 2 to 27. Breed acronyms are reported in <a href="#animals-10-01441-t001" class="html-table">Table 1</a>.</p>
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<p>Neighbor-joining tree constructed on the Reynold’s genetic distance for the breeds considered (<b>A</b>) and based on individual allele-sharing distances (<b>B</b>). Breed acronyms are reported in <a href="#animals-10-01441-t001" class="html-table">Table 1</a>.</p>
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<p>Boxplot of the inbreeding coefficient (F<sub>ROH</sub>) estimated from runs of homozygosity for each breed considered in this study.</p>
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<p>Manhattan plot of each single nucleotide polymorphism (SNP) significance in runs of homozygosity. Blue line indicates the top 0.999% of SNPs.</p>
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10 pages, 950 KiB  
Article
Individual Variability in Response to Social Stress in Dairy Heifers
by Emeline Nogues, Benjamin Lecorps, Daniel M. Weary and Marina A. G. von Keyserlingk
Animals 2020, 10(8), 1440; https://doi.org/10.3390/ani10081440 - 18 Aug 2020
Cited by 16 | Viewed by 4422
Abstract
Regrouping is associated with increased aggression, and disruption of time-budgets. Individuals vary in how well they cope with social stress. Our objective was to describe individual differences in agonistic behavior in dairy heifers after regrouping, and determine how time-budget and behavioral synchronization were [...] Read more.
Regrouping is associated with increased aggression, and disruption of time-budgets. Individuals vary in how well they cope with social stress. Our objective was to describe individual differences in agonistic behavior in dairy heifers after regrouping, and determine how time-budget and behavioral synchronization were affected by these coping strategies. A total of 30 heifers were individually regrouped at 5-months of age into stable groups of 12 unfamiliar animals. For 24 h, agonistic behaviors initiated and received by the regrouped heifer were continuously recorded, and standing, resting and feeding time and synchronization were sampled every 5 min. Scores of engagement in agonistic interactions and avoidance of interactions were calculated for each regrouped heifer. Linear mixed effects models were used to assess whether these two response types were related, and how variation in these responses related to activity and synchronization. Engaged heifers displayed lower avoidance and spent more time feeding. Avoidant heifers spent less time feeding and resting, and were less synchronized while feeding. We conclude that dairy heifers differ in social coping strategy when regrouped through different levels of engagement and avoidance, and that these differences affected their time-budget and behavioral synchronization. Full article
(This article belongs to the Section Cattle)
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<p>Percentage (mean ± SD) of agonistic behaviors initiated and received by dairy heifers during the first 24 h of a regrouping event (n = 30) in relation to: (<b>a</b>) four 6-h periods, (<b>b</b>) the location within the pen where the agonistic behavior was observed.</p>
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<p>The relationship between avoidance and engagement levels (log transformed) of dairy heifers (n = 30) regrouped with unfamiliar conspecifics. Agonistic behaviors were recorded continuously during the first 24 h after heifers were introduced into a new group. More engaged individuals displayed less avoidance (Wald test on a linear mixed-effects model with host group as a random effect; <span class="html-italic">p</span> = 0.02). The extreme point for avoidance was capped in the model at the second highest value + 1.</p>
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<p>The relationship between each regrouped heifer’s avoidance level (n = 30) and the percentage of scans they were recorded feeding or resting (samples were taken every five minutes). Agonistic behaviors were recorded continuously during the first 24 h after heifers were introduced into a new group. More avoidant heifers spent less time feeding (Wald test on a linear mixed-effects model with host group as a random effect; <span class="html-italic">p</span> = 0.0014) and less time resting (<span class="html-italic">p</span> = 0.01). No relation was found between avoidance and time spent resting. The extreme point for avoidance was capped in the model at the second highest value + 1.</p>
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<p>The relationship between each regrouped heifer’s avoidance (n = 30) and behavioral synchronization for feeding or resting. Synchronization was assessed by counting the number of individuals involved in the same activity as the regrouped heifer at each scan, and averaging over the number of scans the heifer was recorded as either feeding or resting. Agonistic behaviors were recorded continuously during the first 24 h after heifers were introduced into a new group. More avoidant heifers were less synchronized while feeding (Wald test on a linear mixed-effects model with host group as a random effect; <span class="html-italic">p</span> = 0.013), but no relationship was found with synchronization while resting (<span class="html-italic">p</span> = 0.36). The extreme point for avoidance was capped in the model at the second highest value + 1.</p>
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18 pages, 1371 KiB  
Article
Histological Evaluation of Gonad Impairments in Russian Sturgeon (Acipenser gueldenstaedtii) Reared in Recirculating Aquatic System (RAS)
by Małgorzata Rzepkowska, Dobrochna Adamek-Urbańska, Magdalena Fajkowska and Marek Łukasz Roszko
Animals 2020, 10(8), 1439; https://doi.org/10.3390/ani10081439 - 18 Aug 2020
Cited by 4 | Viewed by 3686
Abstract
The aim of the study was to raise an issue concerning gonadal impairments in sturgeon reared in recirculating aquatic systems (RAS). In the present study, an in-depth histological evaluation in terms of gonadal pathologies was performed on over-4-year-old (1600 days post-hatching) Russian sturgeon [...] Read more.
The aim of the study was to raise an issue concerning gonadal impairments in sturgeon reared in recirculating aquatic systems (RAS). In the present study, an in-depth histological evaluation in terms of gonadal pathologies was performed on over-4-year-old (1600 days post-hatching) Russian sturgeon (Acipenser gueldenstaedtii) reared under indoor RAS. A female-biased sex ratio, intersex occurrence, ovarian fat overgrowth, T-cell infiltration and follicle atresia were the most commonly observed disorders in the analyzed gonads. The combined processes of oocyte autophagy and follicular cell apoptosis were engaged in follicular atresia; however, atretic follicles showed a varied morphology, whereas oogonia and oocytes in the early stages of meiosis, as well as spermatogonia, underwent degeneration by apoptosis. The most severe pathology was observed in females with abundant intra-ovarian fat deposition. The extremely fatty ovaries were noted to lose the majority of ovarian follicles, which directly leads to fish sterility. The identified impairments might be related to estrogenic endocrine disruption, as feminization and unspecific vitellogenin synthesis were detected, although the sources of the observed pathologies can be diverse. Therefore, the presented research lays the groundwork for further studies on reproductive disorders in this prized and endangered fish species. Full article
(This article belongs to the Special Issue Reproduction of Wild and Cultured Fish)
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<p>Gonads of 1600th-day-post-hatching (dph) Russian sturgeon. (<b>A</b>) Ovary with folded and granular female germinal tissue. (<b>B</b>) Ova-testes with a prevailing ovarian component located in the posterior part of the gonad, and testicular component located in the anterior part of the gonad. (<b>C</b>) Testes-ova with a prevailing testicular component and an ovarian component scattered apically along the gonad surface. (<b>D</b>) Ovary with abundant, white and locally pigmented fat tissue (presence of the germinal tissue on the surface of gonadal folds was only possible to assess by microscopic analyses). Ovarian germinal tissue (white arrowheads), testicular germinal tissue (black arrowheads) and fat (yellow stars).</p>
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<p>Ovaries of 1600th-day-post-hatching (dph) Russian sturgeon. (<b>A</b>) Columnar epithelium in the furrows of the folds. (<b>B</b>) Growing ovarian follicles containing perinucleolar oocytes in primary growth stage. (<b>C</b>) Deposition of vitellogenin (brown) in ooplasm of small perinucleolar oocyte and (<b>D</b>) blood vessel plasma. (<b>E</b>) Nests of oogonia and oocytes at the chromatin-nucleolus stage. (<b>F</b>) Ovarian nests among sparse ovarian follicles. Magnification between (<b>E</b>) and (<b>F</b>): Oogonia and oocytes at the chromatin-nucleolus stage inside the ovarian nest. (<b>G</b>) White fat tissue in the germinal region of the gonad. (<b>H</b>) Final phase of follicular atresia. (<b>I</b>) Residual follicular cells and (<b>J</b>) adipocytes inside a completely phagocytized follicle. (<b>K</b>) Advanced stage of follicular atresia. (<b>L</b>) Normal ovarian follicle (at the top), early stage of follicular atresia (in the middle) and advanced stage of follicular atresia (at the bottom). Magnification inside (<b>L</b>): Heterogeneous ooplasm of degenerating perinucleolar oocyte. (<b>M</b>) Vacuoles and (<b>N</b>) autophagosomic bodies within the ooplasm of degenerating perinucleolar oocytes at an early stage of follicular atresia. (<b>O</b>) Large vesicles in the nuclei of degenerating perinucleolar oocytes at early stage of follicular atresia. (<b>P</b>) Follicular cell apoptosis (brown nuclei). (<b>Q</b>) and magnification inside: Apoptosis of oogonia and chromatin-nucleolus oocytes (brown nuclei). (<b>R</b>) Nest with multiple atretic oogonia and oocytes at the chromatin-nucleolus stage. (<b>S</b>) Small cluster of lymphocytes (<b>T</b>) (dark brown) between ovarian follicles. (<b>T</b>) and magnification inside: Oogonia and primary chromatin-nucleolus stage oocyte nests infiltrated by T- (dark brown) and B-cells (blue). <b>Indicators:</b> secretory cells resembling goblet cells (black arrowhead), adipocytes (black stars), basement membrane (green arrowhead), hypertrophied follicular cells (grey arrowheads), autophagosomic bodies (yellow arrowheads), vacuoles (red arrowheads), prefollicular cells (blue arrowhead), pyknotic nuclei at different stages of oogonia/chromatin-nucleolus oocyte atresia (white arrowhead), B lymphocytes (black arrows), and interstitial and superficial fluid containing leukocytes (red stars). <b>Staining:</b> Masson trichrome (<b>A</b>,<b>B</b>,<b>O</b>), immunohistochemical (IHC)—anti-vitellogenin antibody (<b>C</b>,<b>D</b>), Azan trichrome ((<b>E</b>,<b>F</b>) and magnification in between, (<b>H</b>)), PAS with Weigert’s iron hematoxylin (<b>G</b>,<b>M</b>), AB/PAS (<b>I</b>,<b>J</b>), H-E ((<b>K</b>,<b>L</b>) and magnification inside, (<b>N</b>,<b>R</b>)), TUNEL ((<b>P</b>,<b>Q</b>) and magnification inside), IHC—anti-CD 3 antibody ((<b>S</b>,<b>T</b>) and magnification inside (<b>T</b>)). <b>Scale bars</b>: (<b>A</b>), magnification between (<b>F</b>) and (<b>E</b>,<b>I</b>,<b>M</b>) and magnification inside (<b>Q</b>) = 10 µm; (<b>B</b>–<b>H</b>,<b>L</b>,<b>Q</b>–<b>T</b>) = 100 µm; (<b>J</b>,<b>K</b>,<b>N</b>–<b>P</b>) and magnification inside (<b>L</b>,<b>T</b>) = 50 µm.</p>
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<p>Testis (<b>A</b>,<b>B</b>,<b>P</b>) and intersex gonads (<b>C</b>–<b>O</b>,<b>Q</b>–<b>T</b>) of 1600th-day-post-hatching (dph)Russian sturgeon. (<b>A</b>) Low columnar epithelium on the testicular surface with a thin tunica albuginea and seminiferous tubules underneath. Magnification inside (<b>A</b>): Primary spermatocytes. (<b>B</b>) and magnification inside: Accumulation of spermatozoa (red) inside distal seminiferous tubules. (<b>C</b>) Intersex gonad with ovarian follicles located ventrally (female symbol ♀) and seminiferous tubules located dorsally (male symbol ♂). (<b>D</b>) Chromatin-nucleus oocytes inside seminiferous tubule. (<b>E</b>) Spermatogenesis in cysts of an enlarged seminiferous tubule. (<b>F</b>) Pre-meiotic seminiferous tubules with low spermatogonia density. (<b>G</b>) Apoptotic degeneration of spermatogonia in a pre-meiotic seminiferous tubule. (<b>H</b>) Severe apoptosis in meiotic seminiferous tubules. Magnification inside (<b>H</b>): Apoptotic clusters of spermatogenic cells inside seminiferous tubules. (<b>I</b>,<b>J</b>) Chromatin-nucleus oocytes located in the folded ventral area of the gonad. (<b>K</b>) Increased apoptosis in the feminizing area of the intersex gonad. (<b>L</b>) and magnification within: Infiltration of pre-follicular cells into the female area of intersex gonad. (<b>M</b>) Infiltration of T-cells (dark brown area) into the female area of the intersex gonad. (<b>N</b>) Vasodilation in the male area of the intersex gonad. (<b>O</b>) Seminiferous tubule necrosis in the male area of the intersex gonad. (<b>P</b>) Clusters of T-cells (dark brown) resembling lymph nodules in the germinal area of the testis. (<b>Q</b>) Interstitial white fat tissue located in the ovarian component of the intersex gonad. (<b>R</b>) White fat tissue located laterally to male area of intersex gonad. (<b>S</b>) Vitellogenin (brown) in the plasma of intersex gonad blood vessels. (<b>T</b>) Vitellogenin deposition (brown) in a Sertoli cell adjacent to an ovarian follicle. <b>Indicators:</b> tunica albuginea (white star), spermatogonia (green arrowheads), Sertoli cells (red arrowheads), primary spermatocytes (Sc I), secondary spermatocytes (Sc II), spermatid–spermatozoa conversion (St-Sz), degenerating spermatogonia (blue arrowheads), oocytes (yellow arrowheads), blood vessels (black arrows), foci of necrosis (white arrow), adipocytes (black stars). <b>Staining</b>: Azan trichrome ((<b>A</b>,<b>B</b>) and magnifications inside), H-E ((<b>C</b>–<b>F</b>,<b>I</b>,<b>J</b>,<b>L</b>) and magnification inside, (<b>N</b>,<b>O</b>,<b>Q</b>)), TUNEL ((<b>G</b>,<b>H</b>) and magnification inside, (<b>K</b>)), IHC—anti-CD 3 antibody (<b>M</b>,<b>P</b>), PAS with Weigert’s iron hematoxylin (<b>R</b>), IHC—anti-vitellogenin antibody (<b>S</b>,<b>T</b>). <b>Scale bars</b>: (<b>A</b>–<b>C</b>,<b>H</b>,<b>L</b>,<b>N</b>) = 100 µm; (<b>D</b>–<b>F</b>,<b>G</b>,<b>J</b>,<b>K</b>,<b>O</b>,<b>S</b>,<b>T</b>) = 50 µm; (<b>I</b>,<b>M</b>,<b>P</b>–<b>R</b>) = 500 µm; magnifications inside (<b>A</b>,<b>B</b>,<b>H</b>,<b>L</b>) = 10 µm.</p>
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8 pages, 405 KiB  
Brief Report
Long-Term Effect of Environmental Enrichment on Reproductive Performance of Swiss Webster Mice and Their Female Offspring
by María Noel Meikle, Ana Paula Arévalo, Geraldine Schlapp, Gabriel Fernández-Graña, Alejo Menchaca and Martina Crispo
Animals 2020, 10(8), 1438; https://doi.org/10.3390/ani10081438 - 18 Aug 2020
Cited by 4 | Viewed by 3177
Abstract
The aim of this study was to evaluate the effect of an environmental enrichment (EE) plan on the reproductive performance of Swiss Webster mice and their female offspring used as recipients for embryo transfer. A total of 54 breeder mice and 60 F1 [...] Read more.
The aim of this study was to evaluate the effect of an environmental enrichment (EE) plan on the reproductive performance of Swiss Webster mice and their female offspring used as recipients for embryo transfer. A total of 54 breeder mice and 60 F1 females, used as foster mothers, were allocated in two experimental groups to receive or not receive EE for physical well-being. Reproductive outcomes of the Swiss trios such as birth rate and pup number, litter size, pups’ weight at weaning, interlitter interval and time to first litter were analyzed. Environmental enrichment significantly increased pups weight from breeding trios compared to the control group (14.4 ± 0.1 vs. 13.8g ± 0.1, EE vs. control, respectively; p < 0.01). Other parameters did not differ between both groups. Reproductive parameters of female offspring used as recipients for embryo transfer did not differ among groups subjected or not to EE. These data demonstrate that the EE protocol applied in Swiss Webster breeder mice positively enhanced pups weight, and did not interfere with other reproductive outcomes. In conclusion, this study supports the implementation of EE plans usually applied for animal welfare in mouse facilities with slight improvement in reproductive performance. Full article
(This article belongs to the Special Issue Environmental Enrichment for Animals in our Care)
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<p>Schematic representation of the experimental design. In Experiment 1, Swiss Webster breeder trios were exposed (environmental enrichment (EE) group) or not (control group) to an environmental enrichment (EE) program. During 1-year period a total of 18 trios (nine per experimental group) were compared throughout their reproductive life. Reproductive outcomes such as time to first litter, interlitter interval, birth rate, litter size, number of pups weaned and pup’s weight were registered. In Experiment 2, female offspring that were born from Experiment 1 (i.e., with or without EE) were used as recipients for embryo transfer. Recipient females continued with (EE group, <span class="html-italic">n</span> = 35) or without (control group, <span class="html-italic">n</span> = 25) EE during growth, embryo transfer, pregnancy and delivery until pup’s weaning. Pregnancy and birth rate, litter size number and number of pups weaned were compared between both experimental groups.</p>
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17 pages, 6716 KiB  
Article
Tumor Infiltrating Lymphocytes in Pet Rabbit Mammary Carcinomas: A Study with Relevance to Comparative Pathology
by Sandra Schöniger, Sophie Degner, Qian Zhang, Claudia Schandelmaier, Heike Aupperle-Lellbach, Bharat Jasani and Heinz-Adolf Schoon
Animals 2020, 10(8), 1437; https://doi.org/10.3390/ani10081437 - 17 Aug 2020
Cited by 4 | Viewed by 3240
Abstract
Tumor infiltrating lymphocytes (TILs) serve as prognostic biomarker in human breast cancer. Rabbits have the potential to act as animal model for human breast cancer, and close similarities exist between the rabbit and human immune system. The aim of this study is to [...] Read more.
Tumor infiltrating lymphocytes (TILs) serve as prognostic biomarker in human breast cancer. Rabbits have the potential to act as animal model for human breast cancer, and close similarities exist between the rabbit and human immune system. The aim of this study is to characterize TILs in pet rabbit mammary carcinomas and to statistically correlate results with histological and immunohistochemical tumor characteristics. Microscopic evaluation of TILs was performed in hematoxylin and eosin stained sections of 107 rabbit mammary carcinomas according to international guidelines for human breast cancer. Data on histological features of malignancy, estrogen and progesterone receptor status and calponin expression were obtained from the data base. This study revealed a statistical association between stromal TILs in the central tumor (CT) and infiltrative margin. Higher maximal percentages of stromal TILs at the CT were statistically correlated with decreased mitotic count and lower tumor grade. An increased number of calponin positive tumor cells was statistically associated with a lower mitotic count and a higher percentage of stromal TILs. Results suggest that higher percentages of stromal TILs are useful biomarkers that may point toward a favorable prognosis in rabbit mammary carcinomas and support the concept of the use of rabbits for translational research. Full article
(This article belongs to the Special Issue Biomarkers in Animal Health and Disease)
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<p>Stromal tumor infiltrating lymphocytes (TILs) within the central tumor (CT) of pet rabbit mammary carcinomas. Illustrated are two representative tumor areas with very few stromal TILs (<b>A</b>,<b>A’</b>) and multifocal aggregates of stromal TILs (<b>B</b>,<b>B’</b>), respectively. (<b>A</b>,<b>A’</b>) Grade I carcinoma with a tubular growth pattern and moderate secretory activity. Occasional stromal TILs (less than 1%) are present in the 20× objective field of view (FOV) shown in A. The rectangular area delineated in A is illustrated in A’ in higher magnification. Individual rare stromal TILs are labelled by arrows. (<b>B</b>,<b>B’</b>) Stromal TILs (asterisks) form multifocal aggregates that dissect between tubular structures lined by tumor cells (arrowheads). The 20× objective FOV present in B contains an estimate of 50% stromal TILs. The area delineated by a rectangle is shown in B’ in higher magnification.</p>
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<p>Stromal tumor infiltrating lymphocytes (TILs) within the infiltrative margin (IM) of pet rabbit mammary carcinomas. Shown are two representative tumors with nearly complete absence of stromal TILs (<b>A</b>,<b>A’</b>) and multifocal clusters of stromal TILs (<b>B</b>,<b>B’</b>) at the IM, respectively. (<b>A</b>,<b>A’</b>) Grade II carcinoma with tubular and cystic growth patterns and mild secretory activity. The IM contains no unequivocal stromal TILs. In A, the 10× objective field of view (FOV) is depicted. The rectangle in A delineates the area that is shown in A’ in higher magnification. (<b>B</b>,<b>B’</b>) Grade II carcinoma with a predominantly tubular growth. TILs (green asterisks) are mostly located between and adjacent to the infiltrative tumor cell nests (arrowheads). In B, the 20× objective FOV is shown and contains approximately 25% stromal TILs. The area that is contained within the rectangle is depicted in B’ in close up.</p>
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<p>Intra-tumoral infiltrating lymphocytes (IT TILs) of pet rabbit mammary carcinomas. Depicted are two representative tumors with almost lack of IT TILs (<b>A</b>) and some IT TILs (<b>B</b>), respectively. In comparison to stromal TILs (asterisk), IT TILs (arrowheads) are immediately associated with tumor cells (arrowheads). Intra-tumoral cell fragments (arrows) are present (<b>A</b>,<b>B</b>).</p>
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<p>(<b>A</b>) Of the 107 rabbit mammary carcinomas, 57% were grade I, 41% grade II, and 2% grade III. (<b>B</b>) The majority (65%) was hormone receptor negative, whereas 17% of the tumors expressed both receptors, 17% showed immunostaining for solely progesterone receptor, and 1% was immunoreactive for only estrogen receptor. ERα = estrogen receptor; PR = progesterone receptor.</p>
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<p>(<b>A</b>) Rabbit mammary tumors showed mostly mild (45%) and moderate (43%) invasive behavior; only small percentages of tumors displayed minimal (4%) and marked (8%) tissue invasion. (<b>B</b>) Necrosis was absent in 22% of the tumors, whereas minimal and mild necrosis was detected in 44% and 22% of rabbit mammary carcinomas, respectively. Moderate necrosis was present in 10% of the tumors and 2% of tumors had marked necrosis.</p>
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<p>Higher maximal (max.) stromal tumor infiltrating lymphocytes (TILs) per 20× objective field of view (FOV) within the central tumor (CT) are statistically correlated with decreased numbers of mitotic figures per ten 40× HPFs (<b>A</b>) and a lower histological tumor grade (<b>B</b>).</p>
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<p>An increased percentage of calponin positive tumor cells is statistically correlated with higher maximal (max.) stromal tumor infiltrating lymphocytes (TILs) per 20× objective FOV within the central tumor (CT), (<b>A</b>) and higher average stromal TILs within the central tumor (<b>B</b>).</p>
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15 pages, 1281 KiB  
Article
Rederivation by Cryopreservation of a Paternal Line of Rabbits Suggests Exhaustion of Selection for Post-Weaning Daily Weight Gain after 37 Generations
by Jorge Daniel Juarez, Francisco Marco-Jiménez, Raquel Lavara and José Salvador Vicente
Animals 2020, 10(8), 1436; https://doi.org/10.3390/ani10081436 - 17 Aug 2020
Cited by 7 | Viewed by 2905
Abstract
Rabbit selection programmes have mainly been evaluated using unselected or divergently selected populations, or populations rederived from cryopreserved embryos after a reduced number of generations. Nevertheless, unselected and divergent populations do not avoid genetic drift, while rederived animals seem to influence phenotypic traits [...] Read more.
Rabbit selection programmes have mainly been evaluated using unselected or divergently selected populations, or populations rederived from cryopreserved embryos after a reduced number of generations. Nevertheless, unselected and divergent populations do not avoid genetic drift, while rederived animals seem to influence phenotypic traits such as birth and adult weights or prolificacy. The study aimed to evaluate the effect of a long-term selection for post-weaning average daily weight gain (ADG) over 37 generations with two rederived populations. Specifically, two coetaneous populations were derived from vitrified embryos with 18 generational intervals (R19 and R37), reducing or avoiding genetic drift and environmental and cryopreservation effects. After two generations of both rederived populations (R21 vs. R39 generations), all evaluated traits showed some progress as a result of the selection, the response being 0.113 g/day by generation. This response does not seem to affect the estimated Gompertz growth curve parameters in terms of the day, the weight at the inflexion point or the adult weight. Moreover, a sexual dimorphism favouring females was observed in this paternal line. Results demonstrated that the selection programme had improved ADG without variations in adult body weight but, after 37 generations of selection, this trait seems exhausted. Given the reduction in the cumulative reproductive performance and as a consequence in the selection pressure, or possibly/perhaps due to an unexpected effect, rederivation could be the cause of this weak selection response observed from generation 18 onwards. Full article
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<p>Experimental design: Two experimental progenies were developed from vitrified embryos stored in 2000 (19th generation of selection) and 2015 (37th generation of selection). All rabbits were identified and weighted at weaning and end of the fattening period to calculate the average daily gain. A sample of males and females were weighted weekly from birth to 20 weeks age in F1 y F3 generation to estimate Gompertz curve parameters.</p>
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<p>Growth curves from birth to 20-week-old between F1 rederived populations from R19V and R37V generations. V: Rederived from vitrified embryos. Growth curves were fitted using the Gompertz equation for (<b>A</b>) R19V and R37V generations and (<b>B</b>) sex.</p>
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<p>Growth curves from birth to 20-week-old between F3 filial generations from R21 and R39 generations. V: Rederived from vitrified embryos. Growth curves were fitted using the Gompertz equation for (<b>A</b>) R21 and R39 generations and (<b>B</b>) sex.</p>
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20 pages, 1576 KiB  
Article
Changing Human Behavior to Improve Animal Welfare: A Longitudinal Investigation of Training Laboratory Animal Personnel about Heterospecific Play or “Rat Tickling”
by Megan R. LaFollette, Sylvie Cloutier, Colleen M. Brady, Marguerite E. O’Haire and Brianna N. Gaskill
Animals 2020, 10(8), 1435; https://doi.org/10.3390/ani10081435 - 17 Aug 2020
Cited by 8 | Viewed by 5720
Abstract
Despite evidence for rat tickling’s animal welfare benefits, the technique is rarely implemented in part because of a lack of training. This study’s purpose was to determine the efficacy of online-only or online + hands-on training programs on key outcomes for rat tickling [...] Read more.
Despite evidence for rat tickling’s animal welfare benefits, the technique is rarely implemented in part because of a lack of training. This study’s purpose was to determine the efficacy of online-only or online + hands-on training programs on key outcomes for rat tickling in comparison to a waitlist control condition. After completing a baseline survey, laboratory animal personnel currently working with rats in the United States were semi-randomized to receive online-only training (n = 30), online + hands-on training (n = 34), or waitlist control (n = 32). Participants received further surveys directly after training and 2 months later. Data were analyzed using general linear mixed models. At the 2-month follow-up compared to baseline, both training groups reported increased implementation, self-efficacy, knowledge, and familiarity of rat tickling while only the online + hands-on training participants reported increased control beliefs (while the waitlist group stayed the same). At the 2-month follow-up compared to the waitlist, hands-on training participants reported increased self-efficacy and familiarity with rat tickling. Overall, findings show that both online-only and online + hands-on training can improve key outcomes for rat tickling. Although online + hands-on training is slightly more effective, the interactive online-only training has the potential to improve widescale implementation of a welfare-enhancing technique. Full article
(This article belongs to the Special Issue Laboratory Animals)
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<p>Timeline of procedures and assessment for laboratory animal personnel. All participants were given a baseline survey before group assignment. Then, they were either assigned or randomized to groups and then received training. Immediately after training (or a similar time point for waitlist), participants received a post-training survey. Two months later, they received a final follow-up survey.</p>
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<p>Pictorial tickling procedures. (<b>a</b>): Dorsal contact and pin (standard, validated rat tickling procedure), (<b>b</b>): Dorsal contact only or stroking in the cage, (<b>c</b>): Pin only, (<b>d</b>): Two-handed pin only, (<b>e</b>): Stroking in the hand.</p>
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<p>Implementation and intent to provide rat tickling/correct identification of technique. (<b>a</b>) and (<b>b</b>) This figure shows the highest order significant associations from general linear mixed models that impacted participants current implementation and intent to provide rat tickling. Implementation and intent were measured via a self-report survey. Models were run controlling for potential confounding variables. Both scales display only the range of possible responses. Data is presented as least squares mean ± standard error of the mean. The scale of intent is back transformed from log10. (a) * indicates a significant difference from baseline within treatment group. (b) * indicates a significant difference from baseline. (<b>c</b>) The bottom graph shows the percentage of participants that selected a dorsal contact + pin picture when asked to identify which picture indicates the correct rat tickling technique. ^ indicates a significant difference from waitlist.</p>
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<p>Knowledge, self-efficacy, and familiarity with rat tickling. This figure shows the highest order significant associations from general linear mixed models of (<b>a</b>) knowledge, (<b>b</b>) self-efficacy, and (<b>c</b>) familiarity with rat tickling. Variables were measured via a self-report survey. Models were run controlling for demographic, work, and potential confounding variables. The scales of (b) self-efficacy and (c) familiarity are back transformed from log10. Scales display only the range of possible responses. Data is presented as least squares mean ± standard error of the mean. * indicates a significant difference from baseline within the treatment group. ^ indicates a significant difference from the waitlist within the time point.</p>
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<p>Beliefs about rat tickling. This figure shows the highest order significant associations from general linear mixed models of (<b>a</b>) attitudes, (<b>b</b>) subjective norms, and (<b>c</b>) perceived behavioral control with rat tickling. Variables were measured via a self-report survey. Models were run controlling for potential confounding variables. Scales display only the range of possible responses. Data is presented as least squares mean ± standard error of the mean. (a &amp; b) * Indicates a significant difference from baseline. (c) * Indicates a significant difference from baseline within the treatment group.</p>
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<p>Trained personnel—benefits and barriers to rat tickling. The most common themes related to benefits (advantages) and barriers (factors making it difficult) to tickle rats reported by 86 laboratory animal personnel in a 2-month follow-up survey. Graphic includes representative quotes. Sub themes and additional representative quotes are presented in <a href="#app1-animals-10-01435" class="html-app">Supplemental Table S2</a>.</p>
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7 pages, 240 KiB  
Article
Effect of a Multi-Carbohydrase and Phytase Complex on the Ileal and Total Tract Digestibility of Nutrients in Cannulated Growing Pigs
by Jia-Cheng Yang, Li Wang, Ya-Kuan Huang, Lei Zhang, Rui Ma, Si Gao, Chang-Min Hu, Jlali Maamer, Cozannet Pierre, Aurélie Preynat, Xin Gen Lei and Lv-Hui Sun
Animals 2020, 10(8), 1434; https://doi.org/10.3390/ani10081434 - 17 Aug 2020
Cited by 6 | Viewed by 2894
Abstract
The current study evaluated the influence of a multi-carbohydrase and phytase complex (MCPC) on the ileal and total tract digestibility of nutrients in growing pigs. A total of eight barrows (initial BW = 30.7 ± 1.1 kg) were surgically fitted with a T-cannula [...] Read more.
The current study evaluated the influence of a multi-carbohydrase and phytase complex (MCPC) on the ileal and total tract digestibility of nutrients in growing pigs. A total of eight barrows (initial BW = 30.7 ± 1.1 kg) were surgically fitted with a T-cannula at the distal ileum and randomly allotted to four groups. The experiment was conducted according to a 4 × 4 Latin square design, each period lasting 10 days. Pigs were fed four experimental diets, which consisted of two basal diets (BD1, low phytate; BD2, high phytate) with or without MCPC containing at least 1800 U xylanase, 6600 U α-arabinofuranosidase, 1244 U β-glucanase, and 1000 U phytase per/kg corn–soybean meal with 15% corn distillers based diet. The high phytate diet reduced (p < 0.05) the apparent ileal digestibility (AID) of crude protein by 1.4% and the apparent total tract digestibility (ATTD) of organic matter, crude protein, and gross energy by 1.7, 2.3, and 1.9%, respectively, and tended to decrease (p = 0.10) the ATTD of Ca by 17.3%, relative to the low phytate diet. The dietary supplementation of the MCPC increased (p < 0.05) the AID of phosphorus (P) and calcium (Ca) by 34.2% and 31.1% for BD1 and 26.7% and 41.3% for BD2, respectively, and increased (p < 0.05) ATTD of crude fat, P, and Ca by 1.4%, 45.6%, and 9.6% for BD1 and 3.1%, 66.0%, and 52.7% for BD2, respectively. The MCPC supplementation did not significantly increase the AID and (or) ATTD of crude protein, organic matter, and starch. In conclusion, the dietary supplementation of the MCPC could improve the AID of P and Ca and the ATTD of crude fat, P, and Ca. Full article
(This article belongs to the Special Issue Feed Safety and Quality Control)
19 pages, 1219 KiB  
Article
Evaluation of the Impact of Behavioral Opportunities on Four Zoo-Housed Aardvarks (Orycteropus afer)
by Jennifer Hamilton, Grace Fuller and Stephanie Allard
Animals 2020, 10(8), 1433; https://doi.org/10.3390/ani10081433 - 17 Aug 2020
Cited by 8 | Viewed by 4271
Abstract
Evaluations of enrichment are critical to determine if an enrichment program is meeting stated goals. However, nocturnal species can present a challenge if their active periods do not align with caretakers’ schedules. To evaluate enrichment for four aardvarks housed with a natural light [...] Read more.
Evaluations of enrichment are critical to determine if an enrichment program is meeting stated goals. However, nocturnal species can present a challenge if their active periods do not align with caretakers’ schedules. To evaluate enrichment for four aardvarks housed with a natural light cycle, we provided seven different enrichment items aimed at fulfilling two behavioral goals: exploring and foraging. We wanted to understand how the aardvarks used enrichment, if enrichment promoted the defined goals, and how enrichment that achieves its goals affects welfare indicators, including rates of pacing and social behaviors, behavioral diversity, and fecal glucocorticoid metabolites. Twenty-minute observations from video were performed three times a night for a total of 224 observed hours. We found significant differences in how the aardvarks used items from the two enrichment goals throughout the night, with foraging enrichment used more than exploring at first and exploring enrichment used more later. We found that items promoted their defined goals, and aardvarks showed no evidence of habituation throughout the eight-week study. The impact on selected welfare indicators provided evidence of potentially positive changes, including increased affiliative and decreased agonistic interactions accompanying increases in goal behaviors. These results contribute to the current knowledge available on the impact of goal-directed behavioral opportunities on zoo animal welfare. Full article
(This article belongs to the Special Issue Evidence-Based Practice in Zoo Animal Management)
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<p>Percent of time spent using enrichment and the rate of enrichment use for each stated enrichment goal by the time of night (mean ± SE; <span class="html-italic">n</span> = 4). PM observations occurred between 21:00 and 22:00, MID observations occurred between 00:30 and 01:30, and AM observations occurred between 04:00 and 05:00. Significant pairwise comparisons (<span class="html-italic">p</span> &lt; 0.05) within each timeframe are noted on the graph with different letters, using uppercase letters for percent of time and lowercase letters for rate per 20 min. Note that the scale for enrichment use changes between the PM, MID, and AM graphs. Pairwise comparisons can be found in <a href="#app1-animals-10-01433" class="html-app">Table S2</a>.</p>
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<p>Percent of time spent using enrichment (<b>a</b>) and the rate of enrichment use (<b>b</b>) for each enrichment item by the time of night (mean ± SE; <span class="html-italic">n</span> = 4). PM observations occurred between 21:00 and 22:00, MID observations occurred between 00:30 and 01:30, and AM observations occurred between 04:00 and 05:00. Significant pairwise comparisons (<span class="html-italic">p</span> &lt; 0.05) within each timeframe are noted on the graph with different letters, using uppercase letters for percent of time and lowercase letters for rate per 20 min. Note that the scale for enrichment use changes between the PM, MID, and AM graphs. Pairwise comparisons can be found in <a href="#app1-animals-10-01433" class="html-app">Table S4</a>.</p>
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<p>Percent of total time foraging when each enrichment item was available (mean ± SE; <span class="html-italic">n</span> = 4). Significant pairwise comparisons (<span class="html-italic">p</span> &lt; 0.05) are noted on the graph with different uppercase letters. Pairwise comparisons can be found in <a href="#app1-animals-10-01433" class="html-app">Table S5</a>.</p>
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<p>Percent of total time investigating non-food items when each enrichment item was available (mean ± SE; <span class="html-italic">n</span> = 4). Significant pairwise comparisons (<span class="html-italic">p</span> &lt; 0.05) are noted on the graph with different uppercase letters. Pairwise comparisons can be found in <a href="#app1-animals-10-01433" class="html-app">Table S6</a>.</p>
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<p>Percent of time locomoting when each enrichment item was available (mean ± SE; <span class="html-italic">n</span> = 4). Significant pairwise comparisons (<span class="html-italic">p</span> &lt; 0.05) are noted on the graph with different uppercase letters. Pairwise comparisons can be found in <a href="#app1-animals-10-01433" class="html-app">Table S7</a>.</p>
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<p>Depiction of percent of time (<b>a</b>) and rate (<b>b</b>) of using each enrichment item throughout the study (mean ± SE; <span class="html-italic">n</span> = 4). Significant pairwise comparisons (<span class="html-italic">p</span> &lt; 0.05) within each enrichment item are noted on the graph with different letters, using uppercase letters for percent of time and lowercase letters for rate per 20 min. Pairwise comparisons between enrichment items are not shown. Pairwise comparisons can be found in <a href="#app1-animals-10-01433" class="html-app">Table S9</a>.</p>
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19 pages, 4501 KiB  
Article
Investigation of the Prevalence, Virulence Genes, and Antibiogram of Motile Aeromonads Isolated from Nile Tilapia Fish Farms in Egypt and Assessment of their Water Quality
by Fatma A. El-Gohary, Eman Zahran, Eman A. Abd El-Gawad, Adel H. El-Gohary, Fatma M. Abdelhamid, Amany El-Mleeh, Ehab Kotb Elmahallawy and Mona Mohieldin Elsayed
Animals 2020, 10(8), 1432; https://doi.org/10.3390/ani10081432 - 16 Aug 2020
Cited by 34 | Viewed by 4746
Abstract
The aquaculture industry is a fast-growing sector in Egypt; however, the progress of this industry is impeded by many challenges such as poor water quality and associated bacterial infections. Among others, Motile Aeromonas Septicemia (MAS), caused by aeromonads, is among the most important [...] Read more.
The aquaculture industry is a fast-growing sector in Egypt; however, the progress of this industry is impeded by many challenges such as poor water quality and associated bacterial infections. Among others, Motile Aeromonas Septicemia (MAS), caused by aeromonads, is among the most important bacterial diseases affecting aquaculture due to its zoonotic potential. In the present work, motile aeromonads were isolated from water samples (n= 8) and Nile tilapia (n= 240) in four fish farms (farms I, II, III, and IV) in Kafr El-Sheikh province during the period March to August 2017. This step was followed by investigation of the prevalence and phenotypic, molecular, and histopathological characterization of aeromonads. In addition, antimicrobial susceptibility and virulence gene detection were analyzed. Interestingly, physicochemical water analysis revealed different ranges in relation to the fish farms and seasons. More importantly, Aeromonas isolates were phenotypically identified in 33.3% and 12.5% from fish and water samples, respectively. The highest prevalence of motile aeromonads (46.7%) was recorded from farm IV, and only 12.5% of water samples were positive for them. Out of 80 isolates, 65 (81.25%) were molecularly identified at the genus level using gyrase B (gyrB). The prevalence of the virulence genes detected in the isolated motile aeromonads was aerolysin (aer), 52.2%; elastase (ahp), 26.25%; hemolysin (hyl), 35%; and lipase (lip), 3.75%. The antibiogram profile revealed that the highest resistance of aeromonads isolates (80%) was recorded to chloramphenicol, kanamycin, and azithromycin. Meanwhile, lower resistance levels of 40%, 30%, and 20% were found for streptomycin, cefotaxime, and amoxicillin, respectively. The multiple antibiotic resistance (MAR) index values ranged between 0.27 and 0.82 of motile aeromonads isolates. Furthermore, the histopathological examinations of naturally diseased tilapia revealed widespread hepatocellular necrosis with diffuse, numerous rod-shaped bacteria in liver with melanomacrophages and lymphocytic depletion with edema and hemosiderosis in the spleen. Our findings provide an updated epidemiological baseline for future reference and highlight the likely role of the adverse impact of water quality in the outbreaks of motile aeromonads with special reference to virulence genes and antibiotic resistant traits. Full article
(This article belongs to the Special Issue Fish Pathology)
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<p>Histopathological lesions in liver of naturally diseased Nile tilapia showing (<b>a</b>) hepatocellular dissociation (thin arrow) with many necrotic and apoptotic cells (thick arrow). Few rod-shaped bacteria are inside a hepatocyte cytoplasmic vacuole (green arrows). H&amp;E (hematoxylin and eosin), 400×; (<b>b</b>) Widespread hepatocellular necrosis (black arrows) with diffuse, numerous rod-shaped bacteria (green arrows). H&amp;E, 400×; (<b>c</b>) Diffuse, severe necrotizing hepatitis. See widespread, severe hepatocellular lysis (arrows) with few bacteria (green arrow) and widespread infiltrations with numerous macrophages and lymphocytes (thick arrow). H&amp;E, 400×; (<b>d</b>) Hepatopancreatitis with edema widely separated lytic hepatocytes (arrows) and lysis of hepatopancreas (green arrow) and focal aggregations of inflammatory cells (thick arrow).</p>
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<p>Histopathological lesions in spleen of naturally diseased Nile tilapia showing (<b>a</b>) Activation of melanomacrophages (thick arrow) with lymphocytic depletion (arrow), H&amp;E, 400×. (<b>b</b>) Congested blood vessels (thin arrow) with edema widely separated splenic cells (thick arrow) and hemosiderosis (green arrows), H&amp;E, 100×.</p>
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18 pages, 819 KiB  
Article
Isoflurane and Carbon Dioxide Elicit Similar Behavioral Responses in Rats
by Satyajit Kulkarni and Debra Hickman
Animals 2020, 10(8), 1431; https://doi.org/10.3390/ani10081431 - 16 Aug 2020
Cited by 5 | Viewed by 4266
Abstract
Euthanasia in rodents is an ongoing topic of debate due to concerns regarding the aversive nature of gases with anesthetic properties such as carbon dioxide (CO2) and isoflurane. The aim of this study was to expand upon previously published work evaluating [...] Read more.
Euthanasia in rodents is an ongoing topic of debate due to concerns regarding the aversive nature of gases with anesthetic properties such as carbon dioxide (CO2) and isoflurane. The aim of this study was to expand upon previously published work evaluating the aversiveness of CO2 by introducing an isoflurane treatment group in parallel. Aversion was tested using a forced exposure setup and an aversion-avoidance setup. In the first part of the study, 12 naïve female Sprague–Dawley rats were exposed during four consecutive days, once to each of four treatments: isoflurane, fox urine, oxygen, and CO2. In the second part of the study, 24 naïve female Sprague–Dawley rats and 12 rats from the first experiment were exposed to CO2, isoflurane, or both gases. In the forced exposure study, there were no significant differences between CO2 and isoflurane treatments except in line crosses. Overall, rats were more active in the isoflurane and CO2 treatments compared to the control groups, suggesting that isoflurane and CO2 are similarly aversive. In the aversion-avoidance study, rats previously exposed to isoflurane left the dark chamber significantly earlier compared to naïve rats during exposure to isoflurane. We also show that learned aversion to isoflurane is sustained for at least 15 days after initial exposure. Given this result, we suggest that CO2 is superior to isoflurane when euthanizing rodents with prior exposure to isoflurane. Overall, these results confirm previous studies which suggest that care should be taken when considering the serial use of isoflurane as an anesthetic. Full article
(This article belongs to the Special Issue The Welfare of Laboratory Animals)
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<p>Schematic of the chamber used for the forced exposure experiment.</p>
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<p>Schematic of the chamber used for the aversion-avoidance experiment.</p>
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<p>Naïve rats (<b>dark bar</b>) vs. previously exposed rats (<b>light bar</b>) in natural log time to leave the dark chamber when comparing within treatment groups. An asterisk (* or **) signifies when the <span class="html-italic">p</span> value is significant (<span class="html-italic">p</span> &lt; 0.05).</p>
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18 pages, 918 KiB  
Review
A Review of Welfare Indicators of Indoor-Housed Dairy Cow as a Basis for Integrated Automatic Welfare Assessment Systems
by Lisette M.C. Leliveld and Giorgio Provolo
Animals 2020, 10(8), 1430; https://doi.org/10.3390/ani10081430 - 15 Aug 2020
Cited by 26 | Viewed by 6445
Abstract
For on-farm welfare assessment many automatic methods have been developed to detect indicators of reduced welfare. However, there is still a need to integrate data from single sources to obtain a complete picture of the welfare of an animal. This review offers a [...] Read more.
For on-farm welfare assessment many automatic methods have been developed to detect indicators of reduced welfare. However, there is still a need to integrate data from single sources to obtain a complete picture of the welfare of an animal. This review offers a basis for developing integrated automatic systems to assess dairy cow welfare by providing an overview of the main issues that challenge cow welfare (e.g., lameness) and of well-established indicators that could detect these issues on the farm. Based on a literature review of 4 reviews on cow welfare in general and 48 reviews on single welfare issues, we identified 18 different major welfare issues and 76 matching indicators that could be detected automatically on the farm. Several indicators, e.g., feed intake, showed a consistent association with welfare across many different issues. Although some of these indicators are discussed critically, this means there are many indicators that potentially could detect reduced welfare in general. Other types of indicators could detect one specific welfare issue, e.g., increased respiratory rate for heat stress. These different types of indicators combined provide a basis to develop integrated automatic systems that ultimately would help farmers to detect welfare problems at an early stage. Full article
(This article belongs to the Special Issue Dairy Cow Health and Welfare)
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<p>Indicators that have a significant (binomial test, <span class="html-italic">p</span> &lt; 0.05) direction of association with single cow welfare issues (middle). Indicators are categorized as either environment-based (left) or animal-based (right). The welfare issues are categorized according to the welfare concern they relate primarily too. If the indicator has no similar association (in terms of direction) with other welfare issues, the association is indicated by a straight line (specific). If a similar association occurs also with another welfare issue, the association is indicated by a dotted line (non-specific). Positive associations are indicated by green lines and negative associations by red lines.</p>
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21 pages, 3436 KiB  
Article
Effect of Dietary Supplementation with Coarse or Extruded Oat Hulls on Growth Performance, Blood Biochemical Parameters, Ceca Microbiota and Short Chain Fatty Acids in Broiler Chickens
by Deborah Adewole
Animals 2020, 10(8), 1429; https://doi.org/10.3390/ani10081429 - 15 Aug 2020
Cited by 10 | Viewed by 3752
Abstract
The purpose of this study was to determine the effect of dietary supplementation with coarse or extruded oat hulls on growth performance, blood biochemistry, cecal microbiota, and short chain fatty acids (SCFA) in broiler chickens. Chickens were randomly allotted to four dietary treatments [...] Read more.
The purpose of this study was to determine the effect of dietary supplementation with coarse or extruded oat hulls on growth performance, blood biochemistry, cecal microbiota, and short chain fatty acids (SCFA) in broiler chickens. Chickens were randomly allotted to four dietary treatments consisting of a corn-wheat-soybean meal-based diet (Basal), Basal + Bacitracin methylenedisalicylate (BMD), Basal +3% coarse OH (COH), and basal +3% extruded OH (EOH). Feed intake (FI), body weight gain (BWG), and feed conversion ratio (FCR) were recorded weekly. On day 36, eight chickens/treatment were euthanized, blood samples were collected, and organ weights were determined. Cecal digesta samples were collected for the determination of SCFA concentration and microbial DNA sequence. Data were subjected to ANOVA using the mixed procedure of SAS. Alpha diversity was estimated with the Shannon index, and the significance of diversity differences was tested with ANOVA. Birds fed COH and EOH had reduced (p < 0.05) BWG, but there was no effect of treatment on FCR. There was a significant increase (p = 0.0050) in relative gizzard empty weight among birds that were fed COH, compared to the other treatments. Dietary treatments had no effect on blood biochemical parameters and SCFA concentration. Cecal microbial composition of chickens was mostly comprised of Firmicutes and Tenericutes. Seven OTUs that were differentially abundant among treatments were identified. In conclusion, supplementation of broiler chickens’ diets with 3% COH or EOH did not affect the FCR, blood biochemical parameters and SCFA concentration, but modified few cecal microbiota at the species level. Dietary supplementation with COH but not EOH significantly increased the relative gizzard weight. Full article
(This article belongs to the Special Issue Gut Health in Poultry)
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<p>Box-and-whisker plot illustrating the total number of quality filtered per sample. These reads reflect the total number of high-quality sequences that align with 16Sv4, clustered into OTUs, and were assigned a taxonomic classification. BAS = basal diet; BMD = antibiotic diet; COH = diet supplemented with 3% coarse oat hulls; and EOH = diet supplemented with 3% extruded oat hulls.</p>
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<p>Alpha diversity (Shannon index) was computed and illustrated for each sample. BAS = basal diet; BMD = antibiotic diet; COH = diet supplemented with 3% coarse oat hulls; and EOH = diet supplemented with 3% extruded oat hulls.</p>
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<p>pCoA ordination performed by Bray-Curtis dissimilarities showing the microbiome composition for all specimens. BAS = basal diet; BMD = antibiotic diet; COH = diet supplemented with 3% coarse oat hulls; and EOH = diet supplemented with 3% extruded oat hulls.</p>
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<p>Illustration of the mean and standard error of the relative abundances of the most abundant Phyla. BAS = basal diet; BMD = antibiotic diet; COH = diet supplemented with 3% coarse oat hulls; and EOH = diet supplemented with 3% extruded oat hulls.</p>
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<p>Illustration of the mean and standard error of the relative abundances of the most abundant species-level taxa within the four most abundant Phyla. The groupings along the y-axis represent the column of metadata. BAS = basal diet; BMD = antibiotic diet; COH = diet supplemented with 3% coarse oat hulls; and EOH = diet supplemented with 3% extruded oat hulls.</p>
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<p>Illustration of the explanatory taxa for g_Turicibacter (<b>A</b>), f_Lachnospiracea (<b>B</b>), g_Coprococcus (<b>C</b>), and o_RF39_unclassified (<b>D</b>). BAS = basal diet; BMD = antibiotic diet; COH = diet supplemented with 3% coarse oat hulls; and EOH = diet supplemented with 3% extruded oat hulls.</p>
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<p>Illustration of the explanatory taxa for p_Firmicutes (<b>A</b>), o_Clostridilaes (<b>B</b>), and k_Bacteria_unclassified (<b>C</b>). BAS = basal diet; BMD = antibiotic diet; COH = diet supplemented with 3% coarse oat hulls; and EOH = diet supplemented with 3% extruded oat hulls.</p>
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18 pages, 689 KiB  
Article
Productive and Qualitative Traits of Amaranthus Cruentus L.: An Unconventional Healthy Ingredient in Animal Feed
by Fabio Gresta, Giorgia Meineri, Marianna Oteri, Carmelo Santonoceto, Vittorio Lo Presti, Annalisa Costale and Biagina Chiofalo
Animals 2020, 10(8), 1428; https://doi.org/10.3390/ani10081428 - 14 Aug 2020
Cited by 10 | Viewed by 4555
Abstract
Agronomic traits, oil content, fatty acid composition, antioxidant activity, and total phenolic content were studied on eight A. cruentus accessions cultivated in Southern Italy. A one-way ANOVA model was performed to compare accessions and the Principal Components Analysis was applied to identify patterns [...] Read more.
Agronomic traits, oil content, fatty acid composition, antioxidant activity, and total phenolic content were studied on eight A. cruentus accessions cultivated in Southern Italy. A one-way ANOVA model was performed to compare accessions and the Principal Components Analysis was applied to identify patterns in our dataset and highlight similarities and differences. A. cruentus showed valuable seed yield (0.27 kg/m2, on average) comparable to the main tradition cereals used for animal feeding. Seed-oil composition showed significant differences among the accessions. Data showed a higher lipid content than most cereal grains (from 5.6 to 7.3%). Approximately 60% of fatty acids were unsaturated; linoleic fatty acid ranged from 19 to 34%, oleic acid from 29 to 36%, and alfa-linolenic fatty acid from 0.3 to 0.5%, respectively. The saturated/unsaturated fatty acid ratio ranged from 0.5 to 0.8, the hypocholesterolemic:hypercholesterolaemic ratio from 1.7 to 2.7, the Atherogenic Index from 0.38 to 0.66, the Thrombogenic Index from 0.85 to 1.48, the total phenolic content from 0.14 to 0.36 mg/g seeds, and the antioxidant activity (DPPH) from 0.30 to 0.50. The studied seed-oil composition evidenced A. cruentus as a healthy ingredient for animal feed and consequently, as a possible substitute for traditional cereals. Accessions from Mexico and Arizona emerged for their high qualitative traits. Full article
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<p>Principal component (PC) analysis of the studied amaranth accessions; SFA = Saturated fatty acids; MUFA = Monounsaturated fatty acids; PUFA = Polyunsaturated fatty acids; n3 = n-3 Polyunsaturated fatty acids; n6 = n-6 Polyunsaturated fatty acids; SFA/UFA = Saturated/Unsaturated fatty acid ratio; AI = Atherogenic Index; TI = Thrombogenic Index; HH = hypocholesterolaemic/hypercholesterolaemic ratio; PI = peroxidation index; TPC = Total Phenolic Content expressed as gallic acid equivalents (GAE mg/g seeds); DPPH<sup>•</sup> radical scavenging activity was expressed as EC<sub>50</sub> (concentration of dried extract mg/mL solution).</p>
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16 pages, 596 KiB  
Article
Isolates, Antimicrobial Susceptibility Profiles and Multidrug Resistance of Bacteria Cultured from Pig Submissions in New Zealand
by Christopher B. Riley, Kirsty L. Chidgey, Janis P. Bridges, Emma Gordon and Kevin E. Lawrence
Animals 2020, 10(8), 1427; https://doi.org/10.3390/ani10081427 - 14 Aug 2020
Cited by 6 | Viewed by 3171
Abstract
Data on the scope of bacterial pathogens present and the frequency of antimicrobial resistance (AMR) in New Zealand’s pigs are limited. This study describes bacterial isolates, antimicrobial susceptibility data, and multidrug resistance (MDR; resistance to ≥3 antimicrobial classes) from New Zealand pig submissions. [...] Read more.
Data on the scope of bacterial pathogens present and the frequency of antimicrobial resistance (AMR) in New Zealand’s pigs are limited. This study describes bacterial isolates, antimicrobial susceptibility data, and multidrug resistance (MDR; resistance to ≥3 antimicrobial classes) from New Zealand pig submissions. Porcine test data from June 2003 to February 2016 were obtained from commercial veterinary pathology laboratory records. In total, 470/477 unique submissions resulted in bacterial growth, yielding 779 isolates. Sample type was recorded for 360/477 (75.5%); lung (79/360; 21.9%), faecal (61/360; 16.9%) and intestinal (45/360; 12.5%) were most common. The most common isolates were Escherichia coli (186/779, 23.9%), Actinobacillus pleuropneumoniae (43/779; 5.5%), Streptococcus suis (43/779; 5.5%), unidentified Campylobacter spp. (38/779; 4.9%), alpha haemolytic Streptococci (32/779; 4.1%), coagulase negative Staphylococcus spp. (26/779; 3.3%), and Pasteurella multocida (25/779; 3.2%). Susceptibility results were available for 141/779 (18.1%) isolates from 62/470 (13.2%) submissions. Most were susceptible to trimethoprim-sulphonamide (75/81; 92.6%), but fewer were susceptible to penicillin (37/77; 48.1%), tilmicosin (18/43; 41.9%), or tetracyclines (41/114; 36.0%). No susceptibility data were available for Salmonella spp., Campylobacter spp., or Yersinia spp. isolates. MDR was present in 60/141 (42.6%) isolates. More data on sample submission drivers, antimicrobial drug use, and susceptibilities of important porcine bacterial isolates are required to inform guidelines for prudent antimicrobial use, to reduce their prevalence, human transmission, and to minimise AMR and MDR. Full article
(This article belongs to the Section Pigs)
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<p>Percentage of submissions (<span class="html-italic">n</span> = 475) submitted for bacterial culture and antimicrobial susceptibility testing to commercial veterinary diagnostic laboratories between 2003 and 2016, from pigs in different regions of New Zealand as demarcated on the map. The locations of veterinary laboratories where porcine samples were submitted are indicated as red circles.</p>
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15 pages, 255 KiB  
Article
Can Endocrine Dysfunction Be Reliably Tested in Aged Horses That Are Experiencing Pain?
by Heidrun Gehlen, Nina Jaburg, Roswitha Merle and Judith Winter
Animals 2020, 10(8), 1426; https://doi.org/10.3390/ani10081426 - 14 Aug 2020
Cited by 8 | Viewed by 3442
Abstract
The aim of the present study was to evaluate (i) the effects of different intensities and types of treated pain on the basal concentrations of adrenocorticotropic hormone (ACTH) and cortisol, and (ii) the thyrotropin-releasing hormone (TRH) stimulation test, to determine whether treated pain [...] Read more.
The aim of the present study was to evaluate (i) the effects of different intensities and types of treated pain on the basal concentrations of adrenocorticotropic hormone (ACTH) and cortisol, and (ii) the thyrotropin-releasing hormone (TRH) stimulation test, to determine whether treated pain caused a marked increase of ACTH, which would lead to a false positive result in the diagnosis of pituitary pars intermedia dysfunction (PPID). Methods: Fifteen horses with treated low to moderate pain intensities were part of the study. They served as their own controls as soon as they were pain-free again. The horses were divided into three disease groups, depending on their underlying disease (disease group 1 = colic, disease group 2 = laminitis, disease group 3 = orthopedic problems). A composite pain scale was used to evaluate the intensity of the pain. This pain scale contained a general part and specific criteria for every disease. Subsequently, ACTH and cortisol were measured before and after the intravenous application of 1 mg of TRH. Results: There was no significant difference in the basal or stimulated ACTH concentration in horses with pain and controls, between different pain intensities or between disease groups. Descriptive statistics, however, revealed that pain might decrease the effect of TRH on the secretion of ACTH. There was an increase of ACTH 30 min after TRH application (p = 0.007) in the treated pain group, but this difference could not be statistically confirmed. Measuring the basal ACTH concentration and performing the TRH stimulation test for the diagnosis of PPID seem to be possible in horses with low to moderate pain. Full article
(This article belongs to the Section Animal Welfare)
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13 pages, 1059 KiB  
Article
Genome-Wide Assessment of Runs of Homozygosity in Chinese Wagyu Beef Cattle
by Guoyao Zhao, Tianliu Zhang, Yuqiang Liu, Zezhao Wang, Lei Xu, Bo Zhu, Xue Gao, Lupei Zhang, Huijiang Gao, George E. Liu, Junya Li and Lingyang Xu
Animals 2020, 10(8), 1425; https://doi.org/10.3390/ani10081425 - 14 Aug 2020
Cited by 25 | Viewed by 5071
Abstract
Runs of homozygosity (ROH) are continuous homozygous regions that generally exist in the DNA sequence of diploid organisms. Identifications of ROH leading to reduction in performance can provide valuable insight into the genetic architecture of complex traits. Here, we evaluated genome-wide patterns of [...] Read more.
Runs of homozygosity (ROH) are continuous homozygous regions that generally exist in the DNA sequence of diploid organisms. Identifications of ROH leading to reduction in performance can provide valuable insight into the genetic architecture of complex traits. Here, we evaluated genome-wide patterns of homozygosity and their association with important traits in Chinese Wagyu beef cattle. We identified a total of 29,271 ROH segments from 462 animals. Within each animal, an average number of ROH was 63.36 while an average length was 62.19 Mb. To evaluate the enrichment of ROH across genomes, we initially identified 280 ROH regions by merging ROH events across all individuals. Of these, nine regions containing 154 candidate genes, were significantly associated with six traits (body height, chest circumference, fat coverage, backfat thickness, ribeye area, and carcass length; p < 0.01). Moreover, we found 26 consensus ROH regions with frequencies exceeding 10%, and several regions overlapped with QTLs, which are associated with body weight, calving ease, and stillbirth. Among them, we observed 41 candidate genes, including BCKDHB, MAB21L1, SLC2A13, FGFR3, FGFRL1, CPLX1, CTNNA1, CORT, CTNNBIP1, and NMNAT1, which have been previously reported to be related to body conformation, meat quality, susceptibility, and reproductive traits. In summary, we assessed genome-wide autozygosity patterns and inbreeding levels in Chinese Wagyu beef cattle. Our study identified many candidate regions and genes overlapped with ROH for several important traits, which could be unitized to assist the design of a selection mating strategy in beef cattle. Full article
(This article belongs to the Collection Advances in Cattle Breeding, Genetics and Genomics)
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<p>(<b>a</b>) The total numbers and lengths of runs of homozygosity (ROH) of three classes including Small (0.5 to 1 Mb), Medium (1 to 5 Mb), and Large (&gt;5 Mb) classes. (<b>b</b>) Correlation of total ROH numbers and lengths. The ROH number (<span class="html-italic">y</span>-axis) is plotted against the ROH length (<span class="html-italic">x</span>-axis, i.e., the length of Mb covered by ROH) in each individual.</p>
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<p>Distribution characteristics of ROH on chromosomes in Chinese Wagyu beef cattle. The horizontal axis is the single nucleotide polymorphism (SNP) position, which is ordered by the physical location of the genome; the vertical axis is the ROH frequency at this SNP. The horizontal line in the graph is where the ROH frequency equals 10%.</p>
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<p>Circos plot illustrating ROH regions and association results for eight traits. Association significance test for each of ROH was plotted based on −log<sub>10</sub> (<span class="html-italic">p</span>) values using histograms within the gray inner circle. The ROH frequency was determined by occurrence of ROH with each region within the population. The estimated frequency values were shown in the innermost circle. The outermost circle displays the cattle autosomes. The circles from inside to outside represent association results for body height, body length, chest circumference, fat coverage, backfat thickness, ribeye area, carcass length, and slaughter weight, respectively.</p>
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13 pages, 888 KiB  
Article
C57BL/6J and B6129F1 Embryo Transfer: Unilateral and Bilateral Transfer, Embryo Number and Recipient Female Background Control for the Optimization of Embryo Survival and Litter Size
by Sofia Lamas, Filipa Franquinho, Marlene Morgado, João R. Mesquita, Fátima Gärtner and Irina Amorim
Animals 2020, 10(8), 1424; https://doi.org/10.3390/ani10081424 - 14 Aug 2020
Cited by 5 | Viewed by 5134
Abstract
Embryo transfer (ET) is a common procedure in rodent facilities. Optimizing this technique may help to reduce the number of animals, but little information is available regarding wild type strains and the conditions that affect embryo transfer. To explore this theme, 2-cell C57BL/6J [...] Read more.
Embryo transfer (ET) is a common procedure in rodent facilities. Optimizing this technique may help to reduce the number of animals, but little information is available regarding wild type strains and the conditions that affect embryo transfer. To explore this theme, 2-cell C57BL/6J embryos were transferred after overnight culture of freshly collected zygotes using different conditions: unilateral transfers using a total of 6, 8, 12, 15, 20 and 25 embryos were performed initially; then, this strain was also used for bilateral transfers using a total of 6, 12 and 20 embryos equally divided by the two oviducts. Groups of 25 embryos were not tested for the bilateral technique, since this condition produced the lower success rate when using the unilateral technique and 20 embryos would still represent a large number of embryos. A group of 2-cell B6129F1 embryos was also transferred using unilateral and bilateral ET with 6, 12 and 20 embryos. Crl:CD1(ICR) were used as recipient females for non-reciprocal transfers and C57BL/6J were used to test reciprocal transfers (only tested for six C57BL/6J unilateral transfers). Unilateral transfers using C57BL/6J mice produced higher success rates using six embryos, compared to the other groups transferred unilaterally (p-values between 0.0001 and 0.0267), but the mean number of pups per litter was not different among groups. Bilateral transfer produced higher number of pups when 20 embryos were divided by the two oviducts compared to six (p = 0.0012) or 12 (p = 0.0148) embryos, but with no differences in success rates. No statistical differences were found between the groups of B6129F1, but better results were obtained on bilateral transfers using a total of six embryos. For the strain tested (C57BL/6J), the uterine environment (Crl:CD1(ICR) or C57BL/6J recipient) does not impact the outcome of the technique. These results complement previous work published using genetically engineered mice strains and show that unilateral transfers using low number of embryos (6), produce better outcomes when compared to bilateral or unilateral transfers using more embryos. It also highlights differences between the outcome of bilateral transfers in the two strains tested. A set of historical data of genetically engineered mice at a C57BL/6J background was also included, confirming that lower embryo numbers are related to higher success rates. Together, the outcome of these experiments can be important to reduce the number of recipient and donor females, optimize embryo transfers and improve animal welfare discouraging the use of a more invasive technique. Full article
(This article belongs to the Section Animal Reproduction)
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<p>Experimental design and groups.</p>
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<p>(<b>A</b>) Mean number of pups after unilateral and bilateral transfers using 6, 12 and 20 C57BL/6J embryos. Statistically significant differences were found between 6 embryos unilateral group and 6 and 12 embryos bilateral; (<b>B</b>) success rates for unilateral and bilateral transfers using 6, 12 and 20 embryos. Statistically significant differences were found between 6 embryos bilateral and 20 embryos unilateral and bilateral and between 12 and 20 embryos bilateral. Values are mean ± SEM. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>(<b>A</b>) Mean number of pups after unilateral and bilateral transfer for B6129F1 transfer groups (6, 12 and 20 embryos); (<b>B</b>) success rates for unilateral and bilateral transfer for B6129F1 transfer groups (6, 12 and 20 embryos). Values are mean ± SEM. No statistically significant differences were found between the groups.</p>
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17 pages, 1998 KiB  
Article
Mirtazapine Reduces Adipocyte Hypertrophy and Increases Glucose Transporter Expression in Obese Mice
by Ching-Feng Wu, Po-Hsun Hou, Frank Chiahung Mao, Yao-Chi Su, Ching-Yang Wu, Wei-Cheng Yang, Chen-Si Lin, Hsiao-Pei Tsai, Huei-Jyuan Liao and Geng-Ruei Chang
Animals 2020, 10(8), 1423; https://doi.org/10.3390/ani10081423 - 14 Aug 2020
Cited by 11 | Viewed by 5512
Abstract
Metabolic syndrome is known to engender type 2 diabetes as well as some cardiac, cerebrovascular, and kidney diseases. Mirtazapine—an atypical second-generation antipsychotic drug with less severe side effects than atypical first-generation antipsychotics—may have positive effects on blood glucose levels and obesity. In our [...] Read more.
Metabolic syndrome is known to engender type 2 diabetes as well as some cardiac, cerebrovascular, and kidney diseases. Mirtazapine—an atypical second-generation antipsychotic drug with less severe side effects than atypical first-generation antipsychotics—may have positive effects on blood glucose levels and obesity. In our executed study, we treated male high-fat diet (HFD)-fed C57BL/6J mice with mirtazapine (10 mg/kg/day mirtazapine) for 4 weeks to understand its antiobesity effects. We noted these mice to exhibit lower insulin levels, daily food efficiency, body weight, serum triglyceride levels, aspartate aminotransferase levels, liver and epididymal fat pad weight, and fatty acid regulation marker expression when compared with their counterparts (i.e., HFD-fed control mice). Furthermore, we determined a considerable drop in fatty liver scores and mean fat cell size in the epididymal white adipose tissue in the treated mice, corresponding to AMP-activated protein kinase expression activation. Notably, the treated mice showed lower glucose tolerance and blood glucose levels, but higher glucose transporter 4 expression. Overall, the aforementioned findings signify that mirtazapine could reduce lipid accumulation and thus prevent HFD-induced increase in body weight. In conclusion, mirtazapine may be useful in body weight control and antihyperglycemia therapy. Full article
(This article belongs to the Special Issue Animal Endocrinology and Medicine Research)
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<p>Changes in (<b>A</b>) body weight, (<b>B</b>) body weight gain, (<b>C</b>) food intake (per mouse per week), and (<b>D</b>) daily food efficiency in the control and 10 mg/kg/day mirtazapine-treated mice over the 28 treatment days. We present all data in this figure as mean ± SEMs (n = 10) for both groups. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Changes in absolute weights (g) and body-weight-normalized weights (%) of (<b>A</b>) the liver and (<b>B</b>) the epididymal white adipose tissue (EWAT) in the control and 10 mg/kg/day mirtazapine-treated mice over the 28 treatment days. We present all data in this figure as mean ± SEMs (n = 10) for both groups. ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Changes in (<b>A</b>) plasma triglyceride and (<b>B</b>) aspartate aminotransferase (AST) levels in the control and 10 mg/kg/day mirtazapine-treated mice over the 28 treatment days. All data are presented as mean ± SEMs (n = 10) for both groups. *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>(<b>A</b>) Photomicrographs of H&amp;E-stained liver and EWAT samples (magnification, 400×). Changes in (<b>B</b>) fatty liver scores and (<b>C</b>) adipocyte cellularity in the EWAT in the control and 10 mg/kg/day mirtazapine-treated mice over the 28 treatment days. We present all data in this figure as mean ± SEMs (n = 10) for both groups. ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Changes in (<b>A</b>) intraperitoneal glucose tolerance test (IPGTT) (1 g of glucose/kg body weight), (<b>B</b>) area under the curve (AUC) over 120 min after glucose injection, (<b>C</b>) glucose intolerance criterion (Fisher’s exact test), and (<b>D</b>) serum insulin levels in the control and 10 mg/kg/day mirtazapine-treated mice over the 28 treatment days. We present all data in this figure mean ± SEMs (n = 10) for both groups. * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Changes in (<b>A</b>) homeostasis model assessment-insulin resistance (HOMA-IR) index, (<b>B</b>) insulin sensitivity index (ISI), and (<b>C</b>) GLUT4 expression in the gastrocnemius muscle and FASN and p-AMPK expression the liver in the control and 10 mg/kg/day mirtazapine-treated mice over the 28 treatment days. We present all data in this figure as mean ± SEMs (n = 10) for both groups. * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001.</p>
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10 pages, 216 KiB  
Article
Bone Morphology and Strength in the Mid-Diaphysis of the Humerus and Metacarpus in Dairy Calves Prior to Weaning
by Michaela Gibson, Keren Dittmer, Rebecca Hickson, Penny Back and Chris Rogers
Animals 2020, 10(8), 1422; https://doi.org/10.3390/ani10081422 - 14 Aug 2020
Cited by 8 | Viewed by 2479
Abstract
Calf growth rate in relation to future milk production has been thoroughly studied; however, the observation of growth arrest lines in bones from heifers with humeral fractures has highlighted the need to understand bone growth in relation to calf growth. The aim of [...] Read more.
Calf growth rate in relation to future milk production has been thoroughly studied; however, the observation of growth arrest lines in bones from heifers with humeral fractures has highlighted the need to understand bone growth in relation to calf growth. The aim of this study was to describe the relationship of peripheral quantitative computed tomography (pQCT)-derived measures of bone strength and morphology with gross measurements of size and growth in pre-weaning dairy calves. Liveweight, height, body length, girth and leg length were measured at one, six and twelve weeks of age. At these intervals, the mid-diaphysis of the metacarpus was also scanned in the live animal using pQCT. At six and twelve weeks old, a subset of calves were euthanised and the humerus was collected and scanned at the mid-diaphysis using pQCT. Differences in growth patterns were observed between the metacarpus and humerus over time. Weight was the best predictor for measures of periosteal circumference and stress strain index (R2 = 0.49–0.58) in the metacarpus, and also the best predictor for measures of stress strain index at all ages in the humerus (R2 = 0.94). The strong relationship with weight and bone measures emphasises the need for adequate preweaning nutrition for future bone growth. Full article
(This article belongs to the Section Cattle)
19 pages, 588 KiB  
Review
A Review of Current Knowledge on Staphylococcus agnetis in Poultry
by Gustaw M. Szafraniec, Piotr Szeleszczuk and Beata Dolka
Animals 2020, 10(8), 1421; https://doi.org/10.3390/ani10081421 - 14 Aug 2020
Cited by 23 | Viewed by 5741
Abstract
This review aims to summarize recent discoveries and advancements regarding the characteristics of Staphylococcus agnetis (S. agnetis) and its role in poultry pathology. S. agnetis is an emerging pathogen that was primarily associated with mastitis in dairy cattle. After a presumed [...] Read more.
This review aims to summarize recent discoveries and advancements regarding the characteristics of Staphylococcus agnetis (S. agnetis) and its role in poultry pathology. S. agnetis is an emerging pathogen that was primarily associated with mastitis in dairy cattle. After a presumed host jump from cattle to poultry, it was identified as a pathological agent in broiler chickens (Gallus gallus domesticus), causing lameness induced by bacterial chondronecrosis with osteomyelitis (BCO), septicemia, and valvular endocarditis. Economic and welfare losses caused by lameness are global problems in the poultry industry, and S. agnetis has been shown to have a potential to induce high incidences of lameness in broiler chickens. S. agnetis exhibits a distinct repertoire of virulence factors found in many different staphylococci. It is closely related to S. hyicus and S. chromogenes, hence infections caused by S. agnetis may be misdiagnosed or even undiagnosed. As there are very few reports on S. agnetis in poultry, many facts about its pathogenesis, epidemiology, routes of transmission, and the potential impacts on the poultry industry remain unknown. Full article
(This article belongs to the Section Poultry)
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<p>Colonies of <span class="html-italic">S. agnetis</span> (GenBank: MT231940) on Columbia agar with 5% sheep blood.</p>
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13 pages, 743 KiB  
Article
DNA Footprints: Using Parasites to Detect Elusive Animals, Proof of Principle in Hedgehogs
by Simon Allen, Carolyn Greig, Ben Rowson, Robin B. Gasser, Abdul Jabbar, Simone Morelli, Eric R. Morgan, Martyn Wood and Dan Forman
Animals 2020, 10(8), 1420; https://doi.org/10.3390/ani10081420 - 14 Aug 2020
Cited by 5 | Viewed by 5287
Abstract
The Western European Hedgehog (Erinaceous europaeus) is a nocturnal animal that is in decline in much of Europe, but the monitoring of this species is subjective, prone to error, and an inadequate basis for estimating population trends. Here, we report the [...] Read more.
The Western European Hedgehog (Erinaceous europaeus) is a nocturnal animal that is in decline in much of Europe, but the monitoring of this species is subjective, prone to error, and an inadequate basis for estimating population trends. Here, we report the use of Crenosoma striatum, a parasitic nematode specific to hedgehogs as definitive hosts, to detect hedgehog presence in the natural environment. This is achieved through collecting and sampling the parasites within their intermediate hosts, gastropoda, a group much simpler to locate and sample in both urban and rural habitats. C. striatum and Crenosoma vulpis were collected post-mortem from the lungs of hedgehogs and foxes, respectively. Slugs were collected in two sessions, during spring and autumn, from Skomer Island (n = 21), which is known to be free of hedgehogs (and foxes); and Pennard, Swansea (n = 42), known to have a healthy hedgehog population. The second internal transcribed spacer of parasite ribosomal DNA was used to develop a highly specific, novel, PCR based multiplex assay. Crenosoma striatum was found only at the site known to be inhabited by hedgehogs, at an average prevalence in gastropods of 10% in spring and autumn. The molecular test was highly specific: One mollusc was positive for both C. striatum and C. vulpis, and differentiation between the two nematode species was clear. This study demonstrates proof of principle for using detection of specific parasite DNA in easily sampled intermediate hosts to confirm the presence of an elusive nocturnal definitive host species. The approach has great potential as an adaptable, objective tool to supplement and support existing ecological survey methods. Full article
(This article belongs to the Special Issue Applied Hedgehog Conservation Research)
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<p>The sample size (=number of slugs) required to detect at least one infected slug, given true prevalence from 1% (<span class="html-italic">n</span> = 299) to 25% (<span class="html-italic">n</span> = 11). Higher prevalence omitted for clarity: <span class="html-italic">n</span> declines further to 5 (at 50% prevalence) and 3 (75%).</p>
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17 pages, 1611 KiB  
Review
Metabolic Stress in the Transition Period of Dairy Cows: Focusing on the Prepartum Period
by Osvaldo Bogado Pascottini, Jo L. M. R. Leroy and Geert Opsomer
Animals 2020, 10(8), 1419; https://doi.org/10.3390/ani10081419 - 14 Aug 2020
Cited by 61 | Viewed by 8232
Abstract
All modern, high-yielding dairy cows experience a certain degree of reduced insulin sensitivity, negative energy balance, and systemic inflammation during the transition period. Maladaptation to these changes may result in excessive fat mobilization, dysregulation of inflammation, immunosuppression, and, ultimately, metabolic or infectious disease [...] Read more.
All modern, high-yielding dairy cows experience a certain degree of reduced insulin sensitivity, negative energy balance, and systemic inflammation during the transition period. Maladaptation to these changes may result in excessive fat mobilization, dysregulation of inflammation, immunosuppression, and, ultimately, metabolic or infectious disease in the postpartum period. Up to half of the clinical diseases in the lifespan of high-yielding dairy cows occur within 3 weeks of calving. Thus, the vast majority of prospective studies on transition dairy cows are focused on the postpartum period. However, predisposition to clinical disease and key (patho)physiological events such as a spontaneous reduction in feed intake, insulin resistance, fat mobilization, and systemic inflammation already occur in the prepartum period. This review focuses on metabolic, adaptive events occurring from drying off until calving in high-yielding cows and discusses determinants that may trigger (mal)adaptation to these events in the late prepartum period. Full article
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<p>Schematic illustration of the local (and systemic) changes at the dry-off in dairy cows. (<b>A</b>) Milk cessation occurs abruptly 40 to 60 days before expected calving. (<b>B</b>) When milking is ceased abruptly, the cisternal ducts and alveoli of the udder become engorged, raising the intramammary pressure; these events induce the mammary gland involution process. (<b>C</b>) Mammary glands with lesser engorgement do exhibit an inflammatory response; however, the types and numbers of cells involved in the inflammatory response might be different when milk production at the dry-off is high. Mezzetti et al. [<a href="#B19-animals-10-01419" class="html-bibr">19</a>] dichotomized the milk production at the dry-off as high (≥15 kg milk day) or low (&lt;15 kg milk). Cows producing ≥15 kg of milk per day at the dry-off presented a greater inflammatory response than those producing &lt;15 kg milk.</p>
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<p>This schematic figure questions the origin of the systemic inflammation, lipolysis, and spontaneous reduction in feed intake in the prepartum period. It is largely discussed whether cows stop eating because they suffer from systemic inflammation or whether the reduction in the feed intake is responsible for the decreased energy balance and concomitant systemic inflammation in the prepartum period.</p>
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<p>Schematic illustration of the potential origin of the spontaneous reduction in feed intake in the prepartum period in dairy cows. Based on the studies from our research group [<a href="#B68-animals-10-01419" class="html-bibr">68</a>,<a href="#B69-animals-10-01419" class="html-bibr">69</a>,<a href="#B70-animals-10-01419" class="html-bibr">70</a>,<a href="#B71-animals-10-01419" class="html-bibr">71</a>,<a href="#B72-animals-10-01419" class="html-bibr">72</a>,<a href="#B73-animals-10-01419" class="html-bibr">73</a>], it was demonstrated that over-conditioned cows before calving have a greater adipocyte volume, greater numbers of infiltrated macrophages, and upregulation of the tumor necrosis factor (TNF)-α and interleukin (IL)-6 genes in comparison to normal-conditioned cows. Moreover, it was also demonstrated that the insulin resistance (IR), basal lipolysis, and non-esterified fatty acid (NEFA) concentrations were greater in over- than in normal-conditioned cows. This was often associated with higher lipid accumulation in the livers of over-conditioned cows, with upregulation of the haptoglobin gene. Although we did not measure the feed intake of these cows, we hypothesize that over-conditioning cows is associated with the spontaneous reduction in feed intake in the prepartum period, triggered by their “fat cow syndrome’ in the 1 to 2 weeks before calving.</p>
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15 pages, 1605 KiB  
Article
Hibernation Patterns of the European Hedgehog, Erinaceus europaeus, at a Cornish Rescue Centre
by Kathryn E. South, Kelly Haynes and Angus C. Jackson
Animals 2020, 10(8), 1418; https://doi.org/10.3390/ani10081418 - 14 Aug 2020
Cited by 7 | Viewed by 7566
Abstract
The European hedgehog, Erinaceus europaeus, is frequently admitted to rescue centres in the UK. With many overwintering in captivity, there is cause to investigate hibernation patterns in order to inform and improve husbandry and monitoring protocols. Thirty-five hedgehogs were studied over two [...] Read more.
The European hedgehog, Erinaceus europaeus, is frequently admitted to rescue centres in the UK. With many overwintering in captivity, there is cause to investigate hibernation patterns in order to inform and improve husbandry and monitoring protocols. Thirty-five hedgehogs were studied over two winters. Weight change during hibernation for the first winter was used to test for effects of disturbance on different aspects of hibernation, including total duration, frequency and duration of spontaneous arousals. There was no significant difference between the two winters for any of the four aspects studied. Significant positive correlations demonstrated that weight-loss increased with the duration of the hibernation period and with percent of nights spent asleep, but not with the number of arousal events. Thus, weight-loss appears more strongly associated with the proportion of time spent asleep than with the number of arousal events. This was surprising given the assumed energetic expense of repeated arousal and was potentially due to availability of food during arousals. In contrast with previous studies, larger hedgehogs lost less weight per day than did smaller hedgehogs. They also woke up more often (i.e., had more opportunities to feed), which may explain the unexpected pattern of weight-loss. Hibernatory behaviour in captivity differs from that in the wild, likely because of non-natural conditions in hutches and the immediate availability of food. This study provides a basis for further research into the monitoring and husbandry of hedgehogs such that it can be adapted for each individual according to pre-hibernation weight and behaviour during hibernation. Full article
(This article belongs to the Special Issue Applied Hedgehog Conservation Research)
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<p>Mean (±s.e.) components of hibernation by hedgehogs in Cornwall between the winters of 2015–2016 (<span class="html-italic">n</span> = 21) and 2016–2017 (<span class="html-italic">n</span> = 13); (<b>a</b>) total hibernation period, (<b>b</b>) number of spontaneous arousal events, (<b>c</b>) duration of spontaneous arousal events, (<b>d</b>) interval between spontaneous arousal events i.e., number of consecutive days asleep.</p>
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<p>Correlations between total hibernation period and (<b>a</b>) number of spontaneous arousal events and (<b>b</b>) percentage of nights spontaneously aroused (<span class="html-italic">n</span> = 34).</p>
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<p>Correlations between percent weight-loss and (<b>a</b>) total hibernation period, (<b>b</b>) number of arousal events and (<b>c</b>) percentage of nights spent asleep (<span class="html-italic">n</span> = 34).</p>
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<p>Correlations between pre-hibernation weight (g) of hedgehogs in the winter of 2015/16 (<span class="html-italic">n</span> = 17) and (<b>a</b>) number of arousal events, (<b>b</b>) percent daily weight-loss and (<b>c</b>) actual daily weight-loss (with (<b>b</b>,<b>c</b>) excluding individuals that gained weight).</p>
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18 pages, 2807 KiB  
Article
Characterization of microRNAs during Embryonic Skeletal Muscle Development in the Shan Ma Duck
by Chuan Li, Ting Xiong, Mingfang Zhou, Lei Wan, Suwang Xi, Qiuhong Liu, Yi Chen, Huirong Mao, Sanfeng Liu and Biao Chen
Animals 2020, 10(8), 1417; https://doi.org/10.3390/ani10081417 - 14 Aug 2020
Cited by 13 | Viewed by 3351
Abstract
Poultry skeletal muscle provides high quality protein for humans. Study of the genetic mechanisms during duck skeletal muscle development contribute to future duck breeding and meat production. In the current study, three breast muscle samples from Shan Ma ducks at embryonic day 13 [...] Read more.
Poultry skeletal muscle provides high quality protein for humans. Study of the genetic mechanisms during duck skeletal muscle development contribute to future duck breeding and meat production. In the current study, three breast muscle samples from Shan Ma ducks at embryonic day 13 (E13) and E19 were collected, respectively. We detected microRNA (miRNA) expression using high throughput sequencing following bioinformatic analysis. qRT-PCR validated the reliability of sequencing results. We also identified target prediction results using the luciferase reporter assay. A total of 812 known miRNAs and 279 novel miRNAs were detected in six samples; as a result, 61 up-regulated and 48 down-regulated differentially expressed miRNAs were identified between E13 and E19 (|log2 fold change| ≥ 1 and p ≤ 0.05). Enrichment analysis showed that target genes of the differentially expressed miRNAs were enriched on many muscle development-related gene ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, especially mitogen-activated protein kinase (MAPK) signaling pathways. An interaction network was constructed using the target genes of the differentially expressed miRNAs. These results complement the current duck miRNA database and offer several miRNA candidates for future studies of skeletal muscle development in the duck. Full article
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<p>Overview of sequencing data. (<b>A</b>) Pie chart distribution annotation of the small RNAs at embryonic day 13 (E13) and E19; (<b>B</b>) Length distribution percentage of small RNA sequences in embryonic duck breast muscle. All validated reads of 18–26 nucleotides (nt) for E13 and E19 were assessed for size distribution; (<b>C</b>) Length distribution percentage of unique reads in embryonic duck breast muscle. The validated unique reads of 18–26 nt were counted for size distribution.</p>
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<p>Component analysis of miRNAs in embryonic duck breast muscle. (<b>A</b>) PCA plot of miRNAs expressed in each sample. (<b>B</b>) Distribution of miRNAs on duck reference chromosomes. PEDO01000232.1: an unplaced scaffold of duck genome sequence.</p>
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<p>Differentially expressed miRNAs between embryonic day 13 (E13) and E19. (<b>A</b>) Volcano chart of miRNAs expressed at E19 vs. E13. Red dots represent up-regulated miRNAs, blue dots represent down-regulated miRNAs, gray dots indicate no difference in expression. (<b>B</b>) Numbers of up-regulated and down-regulated miRNAs with <span class="html-italic">p</span> &lt; 0.01 and <span class="html-italic">p</span> &lt; 0.05. (<b>C</b>) Heat map of the differentially expressed miRNAs with <span class="html-italic">p</span> &lt; 0.01 at E19 vs. E13 in breast muscle. (<b>D</b>) Venn diagrams of differentially expressed miRNAs at E19 vs. E13 in embryonic breast muscle.</p>
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<p>qRT-PCR validation of the identification of differentially expressed miRNAs. In all panels, values are presented as mean ± SEM. * <span class="html-italic">p</span> &lt; 0.05; ** <span class="html-italic">p</span> &lt; 0.01; *** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Enrichment analysis of the differentially expressed miRNAs. (<b>A</b>) GO function classification of the target genes of differentially expressed miRNAs. The top 25, 15, and 10 GO terms in biological processes, cellular components, and molecular functions, respectively, are shown. (<b>B</b>) The top 20 enriched KEGG signaling pathways of the target genes of differentially expressed miRNAs. (<b>C</b>) Interaction network of differentially expressed miRNAs and their target genes on MAPK signaling pathways. (<b>D</b>) PPI networks for the target genes of differentially expressed miRNAs with a context score percentile &gt;90 and max energy &lt;−25.</p>
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<p>Dual-luciferase reporter assay validates the interaction between hsa-miR-212-5p and target genes. (<b>A</b>) has-miR-212-5p and its target genes (TargetScan and Miranda were used for the prediction. Target genes with context score percentile &gt;80 and max energy &lt;−18 were shown). (<b>B</b>) Interspecies conservation of miR-212-5p. The potential binding sites on FGFRL1 and IGF2BP1 are shown. The mutated sequence is shown in red. hsa, mmu, bta, cfa, gga, and apl represent <span class="html-italic">Homo sapiens</span>, <span class="html-italic">Mus musculus</span>, <span class="html-italic">Bos taurus</span>, <span class="html-italic">Canis lupus familiaris</span>, <span class="html-italic">Gallus gallus</span>, and <span class="html-italic">Anas platyrhynchos</span>. (<b>C</b>,<b>D</b>) Relative luminescence was measured and calculated after co-transfecting a wild type or mutant sequence of <span class="html-italic">FGFRL1</span> and <span class="html-italic">IGF2BP1</span> with miR-212-5p mimic (or mimic NC) in DF-1. In all panels, values are presented as mean ± SEM. * <span class="html-italic">p</span> &lt; 0.05.</p>
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11 pages, 376 KiB  
Article
Analysis of Genetic Diversity in the Czech Spotted Dog
by Karolína Machová, Anita Kranjčevičová, Luboš Vostrý and Emil Krupa
Animals 2020, 10(8), 1416; https://doi.org/10.3390/ani10081416 - 14 Aug 2020
Cited by 10 | Viewed by 3311
Abstract
Loss off genetic diversity negatively affects most of the modern dog breeds. However, no breed created strictly for laboratory purposes has been analyzed so far. In this paper, we sought to explore by pedigree analysis exactly such a breed—the Czech Spotted Dog (CSD). [...] Read more.
Loss off genetic diversity negatively affects most of the modern dog breeds. However, no breed created strictly for laboratory purposes has been analyzed so far. In this paper, we sought to explore by pedigree analysis exactly such a breed—the Czech Spotted Dog (CSD). The pedigree contained a total of 2010 individuals registered since the second half of the 20th century. Parameters such as the mean average relatedness, coefficient of inbreeding, effective population size, effective number of founders, ancestors and founder genomes and loss of genetic diversity—which was calculated based on the reference population and pedigree completeness—were used to assess genetic variability. Compared to the founding population, the reference population lost 38.2% of its genetic diversity, of which 26% is due to random genetic drift and 12.2% is due to the uneven contribution of the founders. The reference population is highly inbred and related. The average inbreeding coefficient is 36.45%, and the mean average relatedness is 74.83%. The effective population size calculated based on the increase of inbreeding coefficient is 10.28. Thus, the Czech Spotted Dog suffered significant losses of genetic diversity that threaten its future existence. Full article
(This article belongs to the Collection Genetic Diversity in Livestock and Companion Animals)
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<p>Coefficient of inbreeding, correction of inbreeding coefficient to pedigree completeness index (PCI) and number of registrations. Years without any registration were excluded (1988 and 1992).</p>
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25 pages, 770 KiB  
Article
Testing an Animal Welfare Assessment Protocol for Growing-Rabbits Reared for Meat Production Based on the Welfare Quality Approach
by Nadina Botelho, Madalena Vieira-Pinto, Pau Batchelli, Joaquim Pallisera and Antoni Dalmau
Animals 2020, 10(8), 1415; https://doi.org/10.3390/ani10081415 - 13 Aug 2020
Cited by 11 | Viewed by 4424
Abstract
The objective of the present study is to present an animal welfare assessment protocol for growing-rabbits for discussion after its implementation in 32 farms from Spain and Portugal. The protocol comprises the principles of Good Feeding, Good Housing, Good Health and Appropriate Behaviour [...] Read more.
The objective of the present study is to present an animal welfare assessment protocol for growing-rabbits for discussion after its implementation in 32 farms from Spain and Portugal. The protocol comprises the principles of Good Feeding, Good Housing, Good Health and Appropriate Behaviour of the Welfare Quality protocols and includes 36 welfare parameters. Overall, the protocol showed a good capacity for discrimination between farms, with scores ranging 44 to 82 points. The protocol seems reliable for the assessment of animal welfare after proper training of auditors. However, for the criteria social behaviour and other behaviours, further research is needed to ascertain if the methodology and times of observation used are appropriate. Some farms had high mortality rates with a low prevalence of health problems, while others had low mortality rates with high prevalence of health problems due to different managements of culling. The protocol should be improved, to impede farms with high mortality rates but a low prevalence of health issues the day of the audit from obtaining better scores than the second type of farms, by limiting the compensation in key measures. The main points to be solved in the growing-rabbit farms were: to provide more space to the animals; register the number of animals culled accurately; change cervical dislocation for another killing method and provide the farmers training in animal welfare. Full article
(This article belongs to the Special Issue Development and Assessment of Animal Welfare Indicators)
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<p>Space allowance (cm<sup>2</sup> per animal) and height (cm) of the cages in growing-rabbit farms (n = 32). For space allowance, excellent is considered when at least 1500 cm<sup>2</sup> are provided per animal in 90% of the cages and acceptable when this is at least 500 cm<sup>2</sup> per animal. The height of the cage is considered excellent when at least 38 cm are provided in 90% of the cages and acceptable when this is at least 32 cm.</p>
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<p>Mortality rates (in white: culled animals; in black: dead animals or overall mortality rates when % of culled animals were not given by the farmer) in growing-rabbit farms (n = 32) in relation to the classification of the animal welfare protocol, where up to 5% of mortality in at least the last thee months is considered excellent, up to 8% of mortality is considered acceptable, and any other value as not acceptable.</p>
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<p>Overall welfare assessment scores, out of a possible 100, for 32 growing-rabbit farms located in Spain and Portugal.</p>
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