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Animals
Volume 12
Issue 9
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Animals, Volume 12, Issue 9 (May-1 2022) – 174 articles

Cover Story (view full-size image): How ferrets are housed and the environmental enrichment provided can directly impact their health and wellbeing. Through an online survey reaching ferret caretakers from pet, laboratory, zoo, rescue, and working sectors, we describe ferret housing, enrichment, the enrichments ferrets most enjoy and those which may be problematic. From 754 responses, most ferrets were housed socially, and housing varied from single-level cages to free-range housing. Environmental enrichments ferrets reportedly enjoyed most were digging, tunnels, human interaction, and exploration. Scent trails were reported to be among the most enjoyable enrichments but were rarely provided. Problematic enrichment included rubber items, which could be chewed and swallowed, and narrow tunnels and fabrics trapping ferrets. Our results suggest all sectors have room to improve housing and enrichment to better ferret welfare. View this paper
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16 pages, 5700 KiB  
Article
Multiobject Tracking of Wildlife in Videos Using Few-Shot Learning
by Jiangfan Feng and Xinxin Xiao
Animals 2022, 12(9), 1223; https://doi.org/10.3390/ani12091223 - 9 May 2022
Cited by 9 | Viewed by 2804
Abstract
Camera trapping and video recording are now ubiquitous in the study of animal ecology. These technologies hold great potential for wildlife tracking, but are limited by current learning approaches, and are hampered by dependence on large samples. Most species of wildlife are rarely [...] Read more.
Camera trapping and video recording are now ubiquitous in the study of animal ecology. These technologies hold great potential for wildlife tracking, but are limited by current learning approaches, and are hampered by dependence on large samples. Most species of wildlife are rarely captured by camera traps, and thus only a few shot samples are available for processing and subsequent identification. These drawbacks can be overcome in multiobject tracking by combining wildlife detection and tracking with few-shot learning. This work proposes a multiobject-tracking approach based on a tracking-by-detection paradigm for wildlife to improve detection and tracking performance. We used few-shot object detection to localize objects using a camera trap and direct video recordings that could augment the synthetically generated parts of separate images with spatial constraints. In addition, we introduced a trajectory reconstruction module for better association. It could alleviate a few-shot object detector’s missed and false detections; in addition, it could optimize the target identification between consecutive frames. Our approach produced a fully automated pipeline for detecting and tracking wildlife from video records. The experimental results aligned with theoretical anticipation according to various evaluation metrics, and revealed the future potential of camera traps to address wildlife detection and tracking in behavior and conservation. Full article
(This article belongs to the Special Issue Use of Camera Trap for a Better Wildlife Monitoring and Conservation)
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<p>We aimed to obtain a few-shot multiobject-tracking model based on few-shot learning. In this framework, we used a few-shot object detector as the detector and a classification network trained based on the few-shot method as the feature extractor. In addition, we also designed a trajectory-reconstruction module to optimize the tracking result.</p> Full article ">Figure 2
<p>The architecture of our proposed few-shot tracker framework: Few-MOT. It consisted of a detection process and a tracking process. The detection process followed a few-shot object detector that directly regressed the objectness score (def), bounding box location (x,y,w,h), and classification score (cls). The tracking process included a few-shot feature-extraction network (Extractor), a matching module, and a trajectory-reconstruction module. The extractor was responsible for extracting the features of each object clip. The matching module then performed the association of targets between frames, and if they met the reconstruction criteria, they were constructed by the trajectory-reconstruction module. The details of this module will be explained in the methods section.</p> Full article ">Figure 3
<p>Filtering similar background processes: (<b>a</b>) calculating the Hamming distance between pairs of images; (<b>b</b>) sorting them in descending order by similarity; (<b>c</b>) removing the remarkably similar samples to ridding unbalance; (<b>d</b>) selecting the top 60% of reasonable samples, as those that could be subsequently blended for the front and back views.</p> Full article ">Figure 4
<p>Example comparison of the EAOD dataset after elastic distortion. Each target was appropriately distorted without distorting the image. In this way, the diversity of target poses was enriched.</p> Full article ">Figure 5
<p>(<b>a</b>) IoU matching; (<b>b</b>) central point matching.</p> Full article ">Figure 6
<p>Schematic diagram of the division of the central area. The diagram on the left is an abstract representation, where we defined the central area as a fixed-scale area at the boundary of the video screen. The real situation is shown in the diagram on the right.</p> Full article ">Figure 7
<p>Tracks 1 and 2 are the respective tracks recorded for two tigers, with the red flag representing the starting point and the blue flag representing the endpoint.</p> Full article ">Figure 8
<p>Tracking example of a giant panda.</p> Full article ">Figure 9
<p>Tracking example of a golden snub-nosed monkey.</p> Full article ">Figure 10
<p>Tracking sequence before and after frame 30: (<b>a</b>) performance without trajectory-reconstruction module; (<b>b</b>) performance with the trajectory-reconstruction module.</p> Full article ">
11 pages, 9578 KiB  
Article
Prevalence and Genotyping of Anaplasma phagocytophilum Strains from Wild Animals, European Bison (Bison bonasus) and Eurasian Moose (Alces alces) in Poland
by Anna W. Myczka, Stanisław Kaczor, Katarzyna Filip-Hutsch, Michał Czopowicz, Elwira Plis-Kuprianowicz and Zdzisław Laskowski
Animals 2022, 12(9), 1222; https://doi.org/10.3390/ani12091222 - 9 May 2022
Cited by 5 | Viewed by 2257
Abstract
Wild large ungulates, like European bison (Bison bonasus) and Eurasian moose (Alces alces), form an important part of the circulation of Anaplasma phagocytophilum, a Gram-negative, intracellular, tick-transmitted bacterium, in the natural environment. Bison and moose tissue samples were [...] Read more.
Wild large ungulates, like European bison (Bison bonasus) and Eurasian moose (Alces alces), form an important part of the circulation of Anaplasma phagocytophilum, a Gram-negative, intracellular, tick-transmitted bacterium, in the natural environment. Bison and moose tissue samples were subjected to 16S rDNA, groEL and ankA partial gene marker amplification with specific primers using various variants of PCR. Out of 42 examined individuals, Anaplasma sp. were detected in 4/13 Eurasian moose (31%) and 7/29 European bison (24%). In addition, 12 groEL and 5 ankA partial gene positive samples were obtained from the examined animals. The phylogenetic analysis of the groEL partial gene classified samples from European bison to ecotype I, and samples from Eurasian moose to ecotype I and II; the analysis of the ankA partial gene assigned the samples to clusters I and IV. This study extends knowledge about A. phagocytophilum in wild large ungulates in Poland. This is the first report about the occurrence of Anaplasma sp. in one of the largest populations of free living European bison in the world. Our findings confirm that strains of A. phagocytophilum from Bison bonasus and Alces alces may constitute a natural reservoir of pathogenic HGA Anaplasma strains. Full article
(This article belongs to the Special Issue Wildlife Diseases)
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Graphical abstract

Graphical abstract
Full article ">Figure 1
<p>Sampling location in western and central parts of Poland. A—Podlaskie Voivodeship; B—Lublin Voivodeship; C—Subcarpathian Voivodeship; D—Masovian Voivodeship.</p> Full article ">Figure 2
<p>Phylogenetic tree of <span class="html-italic">groEL</span> partial gene (528 bp) haplotypes from <span class="html-italic">Anaplasma phagocytophilum</span>, constructed by Bayesian inference (BI) analysis using MrBayes version 3.2. The HKY + G model was chosen as the best-fitting nucleotide substitution model using JModelTest version 2.1.10 software. The scale bars are proportional to the number of substitutions per site. In bold, sequences from this study. I–IV ecotype of <span class="html-italic">A. phagocytophilum</span> according to Jahfari et al. (2014) [<a href="#B11-animals-12-01222" class="html-bibr">11</a>].</p> Full article ">Figure 3
<p>Phylogenetic tree of ankA partial gene (630 bp) haplotypes from Anaplasma phagocytophilum, constructed by Bayesian inference (BI) analysis using MrBayes version 3.2. The GTR + I + G model was chosen as the best-fitting nucleotide substitution model using JModelTest version 2.1.10 software. The scale bars are proportional to the number of substitutions per site. In bold, sequences from this study.</p> Full article ">
14 pages, 2271 KiB  
Article
Minor Sea Turtle Nesting Areas May Remain Unnoticed without Specific Monitoring: The Case of the Largest Mediterranean Island (Sicily, Italy)
by Oleana Olga Prato, Valentina Paduano, Giulia Baldi, Salvatore Bonsignore, Gerlando Callea, Carlo Camera, Girolamo Culmone, Stefania D’angelo, Diego Fiorentino, Gino Galia, Salvatore Coriglione, Laura Genco, Giuseppe Mazzotta, Nicola Napolitano, Francesco Paolo Palazzo, Giuseppe Palilla, Santo Dylan Pelletti, Toni Mingozzi, Luigi Agresti and Paolo Casale
Animals 2022, 12(9), 1221; https://doi.org/10.3390/ani12091221 - 9 May 2022
Cited by 7 | Viewed by 3864
Abstract
Identifying coastal tracts suitable for sea turtle reproduction is crucial for sea turtle conservation in a context of fast coastal development and climate change. In contrast to nesting aggregations, diffuse nesting is elusive and assessing nesting levels is challenging. A total of 323 [...] Read more.
Identifying coastal tracts suitable for sea turtle reproduction is crucial for sea turtle conservation in a context of fast coastal development and climate change. In contrast to nesting aggregations, diffuse nesting is elusive and assessing nesting levels is challenging. A total of 323 nesting events by the loggerhead sea turtle Caretta caretta have been reported in Sicily, the largest Mediterranean island, in the 1944–2021 period, mostly in the last decade. Specific monitoring efforts are the most likely explanation for such an increase and shows that sea turtle nesting may be underestimated or completely ignored in many areas with scattered nesting. The real nesting level along the 464 km sandy shores of Sicily is still unknown and more research is needed. The observed incubation period was relatively long (57 d) suggesting that a majority of males are produced in Sicily, in contrast to the typical female-biased sex ratio of sea turtles. In a context of climate warming producing sex ratios more skewed towards females, the potential of Sicily as a male-producing area should be further investigated. Other reproductive parameters are provided, such as clutch size and hatching and emergence success. A negative effect of relocation on the latter two was observed. Full article
(This article belongs to the Special Issue Sea Turtles: Ecology, Distribution, and Conservation)
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<p>Sea turtle emergences reported in Sicily from 2017 to 2021 (<b>a</b>), 1944 to 2021 (<b>b</b>) and 1944 to 2016 (<b>c</b>), divided in Nests (stars) and False crawls (circles). Black lines perpendicular to the perimeter of the island indicate the 21 Physiographic Units (PU) identified by letters. The number of nests and the total number of emergences (nests + false crawls) are provided for each PU (in the form nests/emergences) and the totals are given in the top left of the figure. PUs letters are color coded with the Temperature Stations (arrows; Trapani Birgi: TB, Palermo: PA, Ustica Island: US, Boccadifalco: BC, Messina: ME, Capo Rizzuto: CR) that originated the data that were used to calculate IPMT. The circular plot in the middle provides mean annual nest density (nests km-1 year-1) for each PU.</p> Full article ">Figure 1 Cont.
<p>Sea turtle emergences reported in Sicily from 2017 to 2021 (<b>a</b>), 1944 to 2021 (<b>b</b>) and 1944 to 2016 (<b>c</b>), divided in Nests (stars) and False crawls (circles). Black lines perpendicular to the perimeter of the island indicate the 21 Physiographic Units (PU) identified by letters. The number of nests and the total number of emergences (nests + false crawls) are provided for each PU (in the form nests/emergences) and the totals are given in the top left of the figure. PUs letters are color coded with the Temperature Stations (arrows; Trapani Birgi: TB, Palermo: PA, Ustica Island: US, Boccadifalco: BC, Messina: ME, Capo Rizzuto: CR) that originated the data that were used to calculate IPMT. The circular plot in the middle provides mean annual nest density (nests km-1 year-1) for each PU.</p> Full article ">Figure 2
<p>Temporal distribution of loggerhead turtle nests (<span class="html-italic">n</span> = 323) and false crawls (<span class="html-italic">n</span> = 180) recorded in Sicily (Italy) in the 1944–2021 period. Grouped years are reported as in the original source of the data [<a href="#B8-animals-12-01221" class="html-bibr">8</a>].</p> Full article ">Figure 3
<p>Temporal distribution of loggerhead turtle nesting date (DATE) in Sicily (<span class="html-italic">n</span> = 188). IP: incubation period; PIP, IP potentially producing and equal number of males and females; n/a, IP not available.</p> Full article ">Figure 4
<p>Frequency distribution of IP (<span class="html-italic">n</span> = 83), <span class="html-italic">HS</span> (<span class="html-italic">n</span> = 155), ES (<span class="html-italic">n</span> = 155) in loggerhead turtle natural nests in Sicily.</p> Full article ">Figure 5
<p>Frequency distribution of CS (<span class="html-italic">n</span> = 192) and DIST (<span class="html-italic">n</span> = 176) for all loggerhead turtle nests in Sicily.</p> Full article ">Figure 6
<p>Predicted values of IP (estimated marginal means) of loggerhead turtle nests in Sicily plotted separately to show the effect of each IV, originated from the fittest GLM model. Dots in plot (<b>A</b>) show the predicted value for each PU, whiskers represent 95% CI. Plot (<b>B</b>,<b>C</b>) represent regression lines based on the GLM model with 95% CI in light gray. DATE: date of nesting (week of the year). IPMT: mean air temperature during the incubation period (°C).</p> Full article ">
13 pages, 567 KiB  
Article
Partial Substitution of Fish Oil with Microalgae (Schizochytrium sp.) Can Improve Growth Performance, Nonspecific Immunity and Disease Resistance in Rainbow Trout, Oncorhynchus mykiss
by Seunghan Lee, Cheol-Oh Park, Wonsuk Choi, Jinho Bae, Jiyoung Kim, Sera Choi, Kumar Katya, Kang-Woong Kim and Sungchul C. Bai
Animals 2022, 12(9), 1220; https://doi.org/10.3390/ani12091220 - 9 May 2022
Cited by 5 | Viewed by 2747
Abstract
The price of fish oil has reached a historical peak due to a consistent downward production trend, and therefore, the search for sustainable alternative sources has received great attention. This research was conducted to evaluate dietary micro-algae, Schizochytrium sp. (SC) as fish oil [...] Read more.
The price of fish oil has reached a historical peak due to a consistent downward production trend, and therefore, the search for sustainable alternative sources has received great attention. This research was conducted to evaluate dietary micro-algae, Schizochytrium sp. (SC) as fish oil (FO) replacer in rainbow trout, Oncorhynchus mykiss. In the first trial, apparent digestibility coefficient (ADC) was 92.4% for dry matter, 91.4% for crude protein, and 94.2% for crude lipid in rainbow trout. In the second trial, six diets were formulated to replace FO at 0% (CON), 20% (T20), 40% (T40), 60% (T60), 80% (T80), and 100% (T100) with SC in the rainbow trout (3.0 ± 0.4 g, mean ± SD) diet. After eight weeks’ feeding trial, weight gain (WG), specific growth rate (SGR), and feed efficiency (FE) of fish fed the T20 diet were significantly higher than those of fish fed other diets (p < 0.05). However, there were no significant differences in these parameters among those of fish fed CON, T40, T60, and T80 diets. Lysozyme activity of fish fed the T20 diet was significantly higher than those of fish fed other experimental diets (p < 0.05). After 10 days of disease challenge testing with pathogenic bacteria (Lactococcus garvieae 1 × 108 CFU/mL), the cumulative survival rate of fish fed the T20 diet was significantly higher than those of fish fed the CON, T80, and T100 diets. Therefore, these results suggest dietary microalgae SC is well-digested and could replace up to 80% of fish oil in the diet of rainbow trout without negative effects on growth and immune responses. Full article
(This article belongs to the Special Issue Nutritional Management to Promote Responsible Aquaculture)
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<p>Non-specific immune responses of juvenile rainbow trout fed experimental diets for 8 weeks. Values are means from triplicate groups of fish where the values in each row with different superscripts are significantly different (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 2
<p>Cumulative survival rate after challenge with <span class="html-italic">Lactococcus garvieae</span> for 10 days in rainbow trout fed the experimental diets for 8 weeks. <span class="html-italic">p</span>-value: 0.038. Values are means from triplicate groups of fish where asterisks (*) indicates significant differences among groups (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">
9 pages, 2288 KiB  
Article
Effect of a Plateau Environment on the Oxidation State of the Heart and Liver through AMPK/p38 MAPK/Nrf2-ARE Signaling Pathways in Tibetan and DLY Pigs
by Hong Hu, Yongxiang Li, Yuting Yang, Kexing Xu, Lijie Yang, Shiyan Qiao and Hongbin Pan
Animals 2022, 12(9), 1219; https://doi.org/10.3390/ani12091219 - 9 May 2022
Cited by 6 | Viewed by 2143
Abstract
This study evaluated the effect of a plateau environment on the heart and liver oxidation state of Tibetan pigs (TPs) and DLY pigs through analyzing AMPK, p38 MAPK, and Nrf2 signaling pathways. Twelve 120-day-old TPs and twelve 120-day-old DLY pigs were randomly divided [...] Read more.
This study evaluated the effect of a plateau environment on the heart and liver oxidation state of Tibetan pigs (TPs) and DLY pigs through analyzing AMPK, p38 MAPK, and Nrf2 signaling pathways. Twelve 120-day-old TPs and twelve 120-day-old DLY pigs were randomly divided into two groups in a plateau environment for three weeks. Exposed to a plateau environment, TPs exhibited a lower (p < 0.05) malondialdehyde level but higher (p < 0.05) glutathione, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and total antioxidant capacity (T-AOC) activities in the liver and heart than those observed in DLY pigs. TPs also showed higher (p < 0.05) mRNA levels of SOD and GSH-Px in the liver and heart compared with those of DLY pigs. The TPs showed higher (p < 0.05) mRNA and protein levels of AMPK and Nrf2 in the liver and heart compared with those of DLY pigs. Furthermore, TPs showed higher (p < 0.05) mRNA and protein levels of p38 MAPK in the heart and higher mRNA levels of p38 MAPK in the liver compared with those of DLY pigs under a plateau environment. In summary, TPs possess a stronger antioxidant capacity in the heart and liver than that of DLY pigs in a plateau environment through AMPK/p38 MAPK/Nrf2-ARE signaling pathways. Full article
(This article belongs to the Section Animal Physiology)
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<p>Effects of the plateau environment on MDA in heart and liver of TPs and DLY pigs (<span class="html-italic">n</span> = 12). * Significant difference (<span class="html-italic">p</span> &lt; 0.05). MDA = malondialdehyde.</p> Full article ">Figure 2
<p>Effects of the plateau environment on GSH (<b>A</b>), T-AOC (<b>B</b>), GSH-Px (<b>C</b>), SOD (<b>D</b>), and CAT (<b>E</b>) in heart and liver of TPs and DLY pigs (<span class="html-italic">n</span> = 12). * Significant difference (<span class="html-italic">p</span> &lt; 0.05). GSH = glutathione; T-AOC = total antioxidant capacity; GSH-Px = glutathione peroxidase; SOD = superoxide dismutase; CAT = catalase.</p> Full article ">Figure 3
<p>Effects of the plateau environment on mRNA levels of SOD (<b>A</b>), GSH-Px (<b>B</b>), and CAT (<b>C</b>) in heart and liver of TPs and DLY pigs (<span class="html-italic">n</span> = 12). * Significant difference (<span class="html-italic">p</span> &lt; 0.05). SOD = superoxide dismutase; GSH-Px = glutathione peroxidase; CAT = catalase.</p> Full article ">Figure 4
<p>Effects of the plateau environment on protein (<b>A</b>) and mRNA (<b>B</b>) expression levels of AMPK in heart and liver of TPs and DLY pigs (<span class="html-italic">n</span> = 12). * Significant difference (<span class="html-italic">p</span> &lt; 0.05). AMPK = adenosine 5′-monophosphate-activated protein kinase.</p> Full article ">Figure 5
<p>Effects of the plateau environment on protein (<b>A</b>) and mRNA (<b>B</b>) expression levels of p38 MAPK in heart and liver of TPs and DLY pigs (<span class="html-italic">n</span> = 12). * Significant difference (<span class="html-italic">p</span> &lt; 0.05). p38 MAPK = p38 mitogen-activated protein kinase.</p> Full article ">Figure 6
<p>Effects of the plateau environment on protein (<b>A</b>) and mRNA (<b>B</b>) expression levels of Nrf2 in heart and liver of TPs and DLY pigs (<span class="html-italic">n</span> = 12). * Significant difference (<span class="html-italic">p</span> &lt; 0.05). Nrf2 = nuclear factor erythroid 2-related factor 2.</p> Full article ">
12 pages, 1847 KiB  
Article
RNA-Seq Analysis Reveals Expression Regulatory Divergence of W-Linked Genes between Two Contrasting Chicken Breeds
by Hongchang Gu, Liang Wang, Xueze Lv, Weifang Yang, Yu Chen, Kaiyang Li, Jianwei Zhang, Yaxiong Jia, Zhonghua Ning and Lujiang Qu
Animals 2022, 12(9), 1218; https://doi.org/10.3390/ani12091218 - 9 May 2022
Viewed by 1940
Abstract
The regulation of gene expression is a complex process involving organism function and phenotypic diversity, and is caused by cis- and trans- regulation. While prior studies identified the regulatory pattern of the autosome rewiring in hybrids, the role of gene regulation in W [...] Read more.
The regulation of gene expression is a complex process involving organism function and phenotypic diversity, and is caused by cis- and trans- regulation. While prior studies identified the regulatory pattern of the autosome rewiring in hybrids, the role of gene regulation in W sex chromosomes is not clear due to their degradation and sex-limit expression. Here, we developed reciprocal crosses of two chicken breeds, White Leghorn and Cornish Game, which exhibited broad differences in gender-related traits, and assessed the expression of the genes on the W chromosome to disentangle the contribution of cis- and trans-factors to expression divergence. We found that female-specific selection does not have a significant effect on W chromosome gene-expression patterns. For different tissues, there were most parental divergence expression genes in muscle, and also more heterosis compared with two other tissues. Notably, a broader pattern of trans regulation in the W chromosome was observed, which is consistent with autosomes. Taken together, this work describes the regulatory divergence of W-linked genes between two contrasting breeds and indicates sex chromosomes have a unique regulation and expression mechanism. Full article
(This article belongs to the Section Poultry)
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<p><b>Samples and experiment design.</b> Group 1 and Group 2 represent the cross and reciprocal cross, respectively. All analyses use female individuals for two groups in order to identify the expression of the W chromosome. Samples of three tissues (brain, liver, and muscle) were collected from all chicks one day after hatching for transcriptome sequencing.</p> Full article ">Figure 2
<p><b>Hypothetical classification of regulatory divergence.</b> Classification according to the expression level in the maternal parent (mother of CL: Cor/mother of LC: WL), paternal parent (father of CL: WL/father of LC: Cor), and F1 hybrids (CL/LC). Subdivide the ‘Cis’ (<b>a</b>,<b>b</b>) and ‘Trans’ (<b>c</b>–<b>h</b>) main categories are listed. “M”, “F1”, and “P” are the abbreviations for “mother”, “F1 hybrid”, and “female of father breeds” respectively.</p> Full article ">Figure 3
<p><b>Differences in gene expression between Cor and WL.</b> The histogram shows the direction and magnitude of changes in the expression in genes exhibiting divergent parental expression. The two vertical dashed lines represent the expression thresholds of parental divergence. Negative values indicate up-regulated expression of WL, and positive values indicate up-regulated expression of Cor.</p> Full article ">Figure 4
<p><b>Principal Component Analysis using RNA-seq data.</b> (<b>A</b>) PCA results of all three tissues, the sample is divided into 3 different clusters according to the 99% confidence interval. (<b>B</b>) PCA results of the brain. (<b>C</b>) PCA results of the liver. (<b>D</b>) PCA results of muscle. Each dot represents an individual, and dots with different shapes and colors indicate different varieties.</p> Full article ">Figure 5
<p><b>The scatterplot compares the W-linked expressional differences between hybrids and their parental breeds.</b> The visualized results are respectively displayed in (<b>A</b>) all three tissues; (<b>B</b>) brain; (<b>C</b>) liver; and (<b>D</b>) muscle. The different colored dots represent the different regulatory divergence categories. Log<sub>2</sub> Expression F1/F0(maternal) on the X-axis specifically includes two cases, namely log<sub>2</sub> Expression CL/Cor or log<sub>2</sub> Expression LC/WL, corresponding to the two cases on the Y-axis, log<sub>2</sub> Expression CL/WL or log<sub>2</sub> Expression LC/Cor.</p> Full article ">
18 pages, 2293 KiB  
Article
Cardiopulmonary Effects and Pharmacokinetics of Dexmedetomidine Used as an Adjunctive Analgesic to Regional Anesthesia of the Oral Cavity with Levobupivacaine in Dogs
by Matic Pavlica, Mojca Kržan, Ana Nemec, Tina Kosjek, Anže Baš and Alenka Seliškar
Animals 2022, 12(9), 1217; https://doi.org/10.3390/ani12091217 - 9 May 2022
Cited by 3 | Viewed by 2782
Abstract
This study investigated the cardiopulmonary effects and pharmacokinetics of dexmedetomidine (DEX) used as an adjunctive analgesic for regional anesthesia of the oral cavity with levobupivacaine in anesthetized dogs. Forty dogs were randomly assigned to four groups of 10 dogs. All dogs received levobupivacaine [...] Read more.
This study investigated the cardiopulmonary effects and pharmacokinetics of dexmedetomidine (DEX) used as an adjunctive analgesic for regional anesthesia of the oral cavity with levobupivacaine in anesthetized dogs. Forty dogs were randomly assigned to four groups of 10 dogs. All dogs received levobupivacaine (4 blocks) with DEX IO (infraorbital block, n = 10) or IA (inferior alveolar block, n = 10) or placebo (PLC; n = 10) or DEX (n = 10) was injected intravenously (IV) after administration of levobupivacaine. The dose of DEX was always 0.5 µg/kg. Cardiopulmonary parameters were recorded, and blood was drawn for the quantification of DEX in plasma using LC-MS/MS. Heart rate was lower in all LB + DEX groups, while mean arterial pressure (MAP) was higher in the LB + DEX IV and LB + DEX IA groups compared to the LB + PLC IV group. Compared to DEX IV, IO and IA administration resulted in lower MAP up to 2 min after application. Absorption of DEX was faster at IO administration (Cmax and Tmax were 0.47 ± 0.08 ng/mL and 7.22 ± 1.28 min and 0.76 ± 0.09 ng/mL and 7.50 ± 1.63 min for the IO and IA block, respectively). The IA administration resulted in better bioavailability and faster elimination (t1/2 was 63.44 ± 24.15 min and 23.78 ± 3.78 min for the IO and IA block, respectively). Perineural administration of DEX may be preferable because of the less pronounced cardiovascular response compared to IV administration. Full article
(This article belongs to the Special Issue Loco-Regional Anaesthesia in Veterinary Medicine)
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<p>Dogs were randomly assigned to four groups of 10 animals. All dogs were injected with levobupivacaine (all blocks). Dexmedetomidine 0.5 µg/kg was administered into the infraorbital canal (LB + DEX IO), submucosally near the inferior alveolar nerve (LB + DEX IA) or intravenously (LB + DEX IV). The placebo IV (LB + PLC IV) group was administered Sterile Water for Injection IV. The dose of levobupivacaine was calculated as b.w.<sup>0.67</sup> × 0.11 mL/kg for the infraorbital block and b.w.<sup>0.67</sup> × 0.18 mL/kg for the inferior alveolar block. The volume of perineural or IV placebo was equal to the volume of dexmedetomidine. DEX, dexmedetomidine; IA, inferior alveolar block; IO, infraorbital block; IV, intravenously; L, left; LB, levobupivacaine; PLC, placebo (Sterile Water for Injection); R, right.</p> Full article ">Figure 2
<p>Timeline of events. DEX, dexmedetomidine; LB, levobupivacaine; IV, intravenous; MAP, mean arterial pressure; HR, heart rate; <span class="html-italic">f</span><sub>R</sub>, respiratory rate; S, sample; S 0, sample taken prior to administration of oral blocks (B, baseline values); S 1, sample taken immediately after the completion of the last block (BL).</p> Full article ">Figure 3
<p>Concentration-versus-time profile of a single 0.5 µg/kg intravenous dose of dexmedetomidine administered to 10 anesthetized dogs. Dogs were premedicated with methadone intravenously, and anesthesia was induced with propofol and maintained with isoflurane in oxygen. Levobupivacaine was used for both infraorbital and inferior alveolar blocks. The data are presented as mean ± SEM.</p> Full article ">Figure 4
<p>Concentration-versus-time profile of a single 0.5 µg/kg dose of dexmedetomidine injected together with levobupivacaine perineurally into the left (<span class="html-italic">n</span> = 5) or right (<span class="html-italic">n</span> = 5) infraorbital canal in 10 anesthetized dogs. Dogs were premedicated with methadone intravenously, and anesthesia was induced with propofol and maintained with isoflurane in oxygen. Levobupivacaine was used for both infraorbital and inferior alveolar blocks. The data are presented as mean ± SEM.</p> Full article ">Figure 5
<p>Concentration-versus-time profile of a single 0.5 µg/kg dose of dexmedetomidine injected together with levobupivacaine perineurally into the submucosa at the opening of the left (<span class="html-italic">n</span> = 5) or right (<span class="html-italic">n</span> = 5) mandibular foramen in 10 anesthetized dogs. Dogs were premedicated with methadone intravenously, and anesthesia was induced with propofol and maintained with isoflurane in oxygen. Levobupivacaine was used for both infraorbital and inferior alveolar blocks. The data are presented as mean ± SEM.</p> Full article ">Figure 6
<p>Heart rate (HR) before and after administration of dexmedetomidine (DEX) and/or placebo with time points (5, 10, 15, 20, 25 and 30 min) presenting values after completion of IV application of DEX or placebo. Levobupivacaine was used for both infraorbital and inferior alveolar blocks. Values are expressed as median and IQR. B, baseline value, prior to administration of oral blocks; BL, immediately after administration of oral blocks; HR, heart rate; IV1, after one min/completion of IV application of DEX or placebo; IV2, after two min/completion of IV application of DEX or placebo; LB + DEX IA, dogs (<span class="html-italic">n</span> = 10) which received levobupivacaine and dexmedetomidine perineurally into the submucosa at the opening of the mandibular foramen; LB + DEX IO, dogs (<span class="html-italic">n</span> = 10) which received levobupivacaine and dexmedetomidine perineurally into the infraorbital canal; LB + DEX IV, dogs (<span class="html-italic">n</span> = 10) which received levobupivacaine perineurally and dexmedetomidine IV (positive control); LB + PLC IV, dogs (<span class="html-italic">n</span> = 10) which received levobupivacaine perineurally and placebo IV (negative control). * significant difference (<span class="html-italic">p</span> &lt; 0.05) between the LB + PLC IV and LB + DEX IV groups. † significant difference (<span class="html-italic">p</span> &lt; 0.05) between the LB + PLC IV and LB + DEX IA groups. ‡ significant difference (<span class="html-italic">p</span> &lt; 0.05) between the LB + PLC IV and LB + DEX IO groups.</p> Full article ">Figure 7
<p>Mean arterial pressure (MAP) before and after administration of dexmedetomidine (DEX) and/or placebo with time points (5, 10, 15, 20, 25 and 30 min) presenting values after completion of IV application of DEX or placebo. Levobupivacaine was used for both infraorbital and inferior alveolar blocks. Values are expressed as median and IQR. B, baseline value, prior to administration of oral blocks; BL, immediately after administration of oral blocks; IV1, after one min/completion of IV application of DEX or placebo; IV2, after two min/completion of IV application of DEX or placebo; LB + DEX IA, dogs (<span class="html-italic">n</span> = 10) which received levobupivacaine and dexmedetomidine perineurally into the submucosa at the opening of the mandibular foramen; LB + DEX IO, dogs (<span class="html-italic">n</span> = 10) which received levobupivacaine and dexmedetomidine perineurally into the infraorbital canal; LB + DEX IV, dogs (<span class="html-italic">n</span> = 10) which received levobupivacaine perineurally and dexmedetomidine IV (positive control); LB + PLC IV, dogs (<span class="html-italic">n</span> = 10) which received levobupivacaine perineurally and placebo IV (negative control); MAP, mean arterial pressure. * significant difference (<span class="html-italic">p</span> &lt; 0.05) between the LB + PLC IV and LB + DEX IV groups. † significant difference (<span class="html-italic">p</span> &lt; 0.05) between the LB + PLC IV and LB + DEX IA groups. ¢ significant difference (<span class="html-italic">p</span> &lt; 0.05) between the LB + DEX IV and LB + DEX IA groups. ¥ significant difference (<span class="html-italic">p</span> &lt; 0.05) between the LB + DEX IV and LB + DEX IO groups.</p> Full article ">Figure 8
<p>Respiratory rate (<span class="html-italic">f</span><sub>R</sub>) before and after administration of dexmedetomidine (DEX) and/or placebo. Time points (5, 10, 15, 20, 25 and 30 min) present values after completion of IV application of DEX or placebo. Levobupivacaine was used for both infraorbital and inferior alveolar blocks. Values are expressed as median and IQR. B, baseline value, prior to administration of oral blocks; BL, immediately after administration of oral blocks; <span class="html-italic">f</span><sub>R,</sub> respiratory rate; IV1, after one min/completion of IV application of DEX or placebo; IV2, after two min/completion of IV application of DEX or placebo; LB + DEX IA, dogs (<span class="html-italic">n</span> = 10) which received levobupivacaine and dexmedetomidine perineurally into the submucosa at the opening of the mandibular foramen; LB + DEX IO, dogs (<span class="html-italic">n</span> = 10) which received levobupivacaine and dexmedetomidine perineurally into the infraorbital canal; LB + DEX IV, dogs (<span class="html-italic">n</span> = 10) which received levobupivacaine perineurally and dexmedetomidine IV (positive control); LB + PLC IV, dogs (<span class="html-italic">n</span> = 10) which received levobupivacaine perineurally and placebo IV (negative control). * significant difference (<span class="html-italic">p</span> &lt; 0.05) between the LB + PLC IV and LB + DEX IV groups.</p> Full article ">
11 pages, 1538 KiB  
Article
Epidemiology and Genetic Diversity of Spirometra Tapeworm Isolates from Snakes in Hunan Province, China
by Tengfang Gong, Xiaoyi Su, Fen Li, Junlin He, Shuyu Chen, Wenchao Li, Xinrui Xie, Yisong Liu, Xi Zhang and Wei Liu
Animals 2022, 12(9), 1216; https://doi.org/10.3390/ani12091216 - 9 May 2022
Cited by 2 | Viewed by 2009
Abstract
Sparganosis, caused by the plerocercoid larvae of Spirometra tapeworms, is a public health hazard worldwide. The prevalence and genetics of sparganum from snakes remain unclear. In this study, we investigated the prevalence of sparganum infection in wild snakes in Hunan province and compared [...] Read more.
Sparganosis, caused by the plerocercoid larvae of Spirometra tapeworms, is a public health hazard worldwide. The prevalence and genetics of sparganum from snakes remain unclear. In this study, we investigated the prevalence of sparganum infection in wild snakes in Hunan province and compared the prevalence of Spirometra tapeworms in snakes worldwide. Furthermore, the genetic diversity of collected isolates was analyzed using mitochondrial cytb and cox1 genes. The result shows that the sparganum infection rate in wild snakes (89.50%, 402/449) was higher in Hunan than in other regions. Genetic diversity analysis based on concatenated sequences revealed high genetic diversity but no distinct genetic structure among Spirometra populations. Phylogenetic analysis supported the division of European and Chinese Spirometra isolates and a single species in Chinese Spirometra isolates. The prevalence of Spirometra tapeworms in snakes is serious, and the risk of sparganosis should be further publicized. Full article
(This article belongs to the Collection Advances in Parasite Epidemiology and Population Genetics)
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<p>Sampling sites in Hunan province, China. The geographic location is shown in the inset. The sampling sites were added according to GPS data.</p> Full article ">Figure 2
<p>Median-joining network of 67 sequences of <span class="html-italic">cyt</span>b and <span class="html-italic">cox</span>1 genes in sparganum isolates colored by sampling sites (<b>a</b>) and host species (<b>b</b>) in Hunan province, China. The area of circles represents the number of individuals with that haplotype. Perpendicular short lines on the branches indicate unsampled intermediate haplotypes.</p> Full article ">Figure 3
<p>Estimate of demographic expansion of sparganum isolates from Hunan province. (a) Mismatch distribution analyses. The line charts represent the observed frequencies of pairwise differences among haplotypes. (b) Bayesian skyline plot calculations. The <span class="html-italic">X</span>-axis is in units of million years in the past, and the <span class="html-italic">Y</span>-axis is Ne × μ (effective population size × mutation rate per site per generation). The median estimates are shown as thick solid lines, and the 95% HPD limits are represented by the colored areas.</p> Full article ">Figure 4
<p>Phylogenetic relationships among the examined sparganum isolates from different locations in China and Poland were inferred by maximum parsimony (MP) analysis based on the concatenated sequences of <span class="html-italic">cyt</span>b and <span class="html-italic">cox</span>1. The numbers along branches indicate bootstrap values, and bootstrap values above 60 are shown. Circled numbers represent the main clades discussed in the text.</p> Full article ">
22 pages, 1075 KiB  
Article
Social Behavior and Group Formation in Male Asian Elephants (Elephas maximus): The Effects of Age and Musth in Wild and Zoo-Housed Animals
by Chase A. LaDue, Rajnish P. G. Vandercone, Wendy K. Kiso and Elizabeth W. Freeman
Animals 2022, 12(9), 1215; https://doi.org/10.3390/ani12091215 - 8 May 2022
Cited by 10 | Viewed by 4288
Abstract
Asian elephants are endangered, and the long-term viability of the species depends on integrative approaches to address the sustainability of in-situ and ex-situ populations. Growing evidence shows that male elephants exhibit extensive and flexible social behavior that rivals the complexity of that of [...] Read more.
Asian elephants are endangered, and the long-term viability of the species depends on integrative approaches to address the sustainability of in-situ and ex-situ populations. Growing evidence shows that male elephants exhibit extensive and flexible social behavior that rivals the complexity of that of females. Male elephant sociality is expected to change dramatically around the unique sexual state of musth. However, data related to male Asian elephant sociality is lacking. Here, we conducted complementary observations in Wasgamuwa National Park, Sri Lanka, and North American zoos of male Asian elephant social behavior. Age and musth status, along with other factors, were associated with variation in social behavior and group formation of males. In wild male elephants, both musth status and age impacted elephant associations within all-male and mixed-sex groups: non-musth elephants were generally sighted less often in mixed-sex groups as they aged, while the inverse occurred with musth elephants. Musth status interacted with age to predict the number of conspecifics with which a wild male elephant associated: younger males were observed with more females during non-musth (but the opposite was true during musth), and male elephants between 20 and 30 years were observed with the highest number of male conspecifics except during musth. Finally, we found variation in aggression, prosocial behavior, and submissive behavior was influenced by intrinsic (age and musth status) and extrinsic factors (group size and type) in similar ways in both populations; prosocial behavior was most common and was influenced by the number of conspecifics present (both populations), and age, group type, and musth status (zoo population), while aggression was rare, especially among older elephants. We suggest that longitudinal studies of this threatened species will be particularly helpful to promote the reproduction and conservation of Asian elephants in in-situ and ex-situ environments. Full article
(This article belongs to the Special Issue Advances in Elephant Reproduction and Conservation)
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<p>Relative frequencies of group-type sightings (solitary, all-male group, mixed-sex group) as a function of the age class and binary musth status (top panel, non-musth; bottom panel, musth) of the focal male elephants in Wasgamuwa National Park, Sri Lanka. The sample size (number of sightings) for each category is given on the horizontal axes.</p> Full article ">Figure 2
<p>Boxplots of the association of age and binary musth status of male Asian elephants in Wasgamuwa National Park, Sri Lanka, with the number of (<b>a</b>) adult females and (<b>b</b>) adult males in a male’s group. For simplicity, ages and age classes are condensed into younger (10 to 30 years) and older (&gt;30 years) age categories in this plot. Boxes extend from the first to the third quartile, with the median indicated by a thick line; fences extend to 1.5 times the interquartile range, and closed circles indicate values outside this range.</p> Full article ">Figure 3
<p>Relative frequencies of social behavior (aggression, prosocial behavior, dominance behavior, and submissive behavior) between wild and zoo-housed male Asian elephants, separated by social group-type (all-male or mixed sex) and age of focal animal (younger = 10–30 years old, older = 30+ years old). For simplicity, ages and age classes are condensed into younger and older age categories in this plot.</p> Full article ">Figure 4
<p>Boxplot showing the association between age and rates of aggression for wild and zoo-housed male Asian elephants in all-male and mixed-sex groups. For simplicity, ages and age classes are condensed into younger (10 to 30 years) and older (&gt;30 years) age categories in this plot. Boxes extend from the first to the third quartile, with the median indicated by a thick line; fences extend to 1.5 times the interquartile range, and closed circles indicate values outside this range.</p> Full article ">Figure 5
<p>Relationship between rates of prosocial behavior and the number of conspecifics present for wild and zoo-housed male Asian elephants. Closed circles represent individual observation sessions, with regression lines shown for each age class (darker shades represent older age classes). The absence of a regression line for an age class indicates a lack of adequate data for the construction of a relationship. Note difference in scale on horizontal ax is for wild and zoo-housed elephants.</p> Full article ">
18 pages, 1238 KiB  
Article
Impact of the COVID-19 Pandemic on Primate Research and Conservation
by Kim E. Reuter, Seheno Andriantsaralaza, Malene Friis Hansen, Marni LaFleur, Leandro Jerusalinsky, Edward E. Louis, Jonah Ratzimbazafy, Elizabeth A. Williamson and Russell A. Mittermeier
Animals 2022, 12(9), 1214; https://doi.org/10.3390/ani12091214 - 8 May 2022
Cited by 8 | Viewed by 3721
Abstract
There is evidence to suggest that the Coronavirus Disease 2019 (COVID-19) pandemic may hamper our achievement of the Sustainable Development Goals (SDGs). Here, we use non-human primates as a case study to examine the impacts of COVID-19 on the ability to achieve biodiversity [...] Read more.
There is evidence to suggest that the Coronavirus Disease 2019 (COVID-19) pandemic may hamper our achievement of the Sustainable Development Goals (SDGs). Here, we use non-human primates as a case study to examine the impacts of COVID-19 on the ability to achieve biodiversity conservation and management sustainability targets. We collected data through a survey of members of the IUCN SSC Primate Specialist Group from January to March 2022. Of the 93 experts that responded to our survey, we found that 39% had not been able to visit any of their field sites since March 2020, 54% said they had less funding available for their primate-related work, and only one out of ten said they had managed to achieve at least 76–100% of their planned primate-related work since March 2020. Six out of ten respondents (61%) felt that primate conservation efforts in protected areas were worse than before the onset of the COVID-19 pandemic and one-third (33%) felt hunting was happening more frequently than before. This study provides evidence of the impacts of COVID-19 on progress towards achieving the SDGs, and provides practical lessons learned for biodiversity conservation efforts moving forward. Full article
(This article belongs to the Special Issue Primatology and the Achievement of the UN 2030 Sustainable Development Goals)
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<p>Self-reported work productivity by respondents in the past (from March 2020 to March 2022) and into the future (into the next two years). Respondents were asked to estimate how much they had managed to achieve (from four categories) relative to what they would have achieved had the COVID-19 pandemic not occurred.</p> Full article ">Figure 2
<p>Opinions given by respondents on the how the COVID-19 pandemic had affected different aspects of protected area functioning and governance.</p> Full article ">Figure 3
<p>Opinions given by respondents on the how the COVID-19 pandemic had affected different aspects of primate hunting in sites and countries where they worked.</p> Full article ">Figure 4
<p>Opinions given by respondents on the how the COVID-19 pandemic had affected different aspects of pet primate ownership in sites and countries where they worked.</p> Full article ">
10 pages, 3139 KiB  
Article
Liver Receptor homolog-1 Regulates Apoptosis of Bovine Ovarian Granulosa Cells by Progestogen Receptor Signaling Pathway
by Dejun Xu, Xiaohan Jiang, Yukun Wang and Shuaifei Song
Animals 2022, 12(9), 1213; https://doi.org/10.3390/ani12091213 - 8 May 2022
Cited by 1 | Viewed by 1982
Abstract
The purpose of the present investigation was to assess the function of LRH-1 on GCs and the mechanisms involved. Here, LRH- was highly expressed in the bovine GCs of atretic follicles. Treatment with 50 μM of LRH-1 agonist (DLPC) significantly induced the expression [...] Read more.
The purpose of the present investigation was to assess the function of LRH-1 on GCs and the mechanisms involved. Here, LRH- was highly expressed in the bovine GCs of atretic follicles. Treatment with 50 μM of LRH-1 agonist (DLPC) significantly induced the expression of LRH-1 (p < 0.05). In particular, LRH-1 activation blocked the progestogen receptor signaling pathway via downregulating progesterone production and progestogen receptor levels (p < 0.05), but had no effect on 17 beta-estradiol synthesis. Meanwhile, LRH-1 activation promoted the apoptosis of GCs and increased the activity of caspase 3 (p < 0.05). Importantly, upregulating the progestogen receptor signaling pathway with progestogen could attenuate the LRH-1-induced proapoptotic effect. Moreover, treatment with progestogen decreased the activity of the proapoptotic gene caspase 3 and increased the expression of antiapoptotic gene Bcl2 in LRH-1 activated GCs (p < 0.05). Taken together, these results demonstrate that LRH-1 might be dependent on the progestogen receptor signaling pathway to modulate bovine follicular atresia. Full article
(This article belongs to the Special Issue Advances of Endocrinology in Animal Reproduction)
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<p>LRH-1 high expression in the atretic follicles. (<b>A</b>) Immunochemistry assay was performed to visualize the localization of LRH-1 in bovine ovarian cells. Immuno-specific staining was brown, indicating LRH-1-positive cells. Immunohistochemistry was performed on three different ovarian slides from each of three bovines. Atretic follicles were indicated with the red arrow, whereas healthy follicles were indicated with black arrows. Bar: 200 μm. (<b>B</b>) The LRH-1 levels of granulosa cells in atretic follicles or healthy follicles. Data are shown as the means ± SEM of three biological replicates. ** <span class="html-italic">p</span> &lt; 0.01, comparing the indicated groups. TCs, theca cells; GCs, granulosa cells.</p> Full article ">Figure 2
<p>DLPC induced LRH-1 expression. (<b>A</b>) The mRNA levels of LRH-1 in granulosa cells by treatment with various concentrations of LRH-1 agonist (DLPC). (<b>B</b>) Western blotting of LRH-1 protein in granulosa cells by treatment with various concentrations of DLPC. (<b>C</b>) The protein levels of LRH-1 in the different groups. Data are expressed as the mean ± SEM from three biological replicates. * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001, comparing the indicated groups.</p> Full article ">Figure 3
<p>DLPC blocked progestogen receptor signaling pathway. (<b>A</b>) The effect of DLPC on 17 beta-estradiol (E2) production of granulosa cells. The concentration of oestradiol was detected by ELISA assay. (<b>B</b>) The effect of DLPC on progestogen (P4) production of granulosa cells. (<b>C</b>) The mRNA levels of progestogen receptor in granulosa cells by treatment with DLPC. (<b>D</b>) Immunofluorescence of progestogen receptor in granulosa cells by treatment with 50 μM DLPC or/and 10 μM P4. (<b>E</b>) Quantification of progestogen receptor was analyzed with Image J software. Bar: 200 μm. Data are expressed as the mean ± SEM from three independent replicates. Values with different letters (a, b, c) indicate significant differences in bars (<span class="html-italic">p</span> &lt; 0.05). * <span class="html-italic">p</span> &lt; 0.05, *** <span class="html-italic">p</span> &lt; 0.001, comparing the indicated groups.</p> Full article ">Figure 4
<p>DLPC activation promoted apoptosis via progestogen signaling. (<b>A</b>) The viability of granulosa cells was shown by CCK-8 assay in different treatments. (<b>B</b>) Representative images of apoptosis with annexin V-FITC staining by a flow cytometry instrument in 50 μM DLPC-, 50 μM DLPC + 5 μM P4-, 50 μM DLPC + 10 μM P4-, 50 μM DLPC + 100 μM P4-exposed granulosa cells. (<b>C</b>) The percentage of apoptosis in the different treatment groups. (<b>D</b>) Western blotting of Bcl2 and cleaved-caspase 3 in granulosa cells by treatment with 50 μM DLPC, 50 μM DLPC + 1 μM P4, 50 μM DLPC + 5 μM P4, 50 μM DLPC + 10 μM P4 and 50 μM DLPC + 100 μM P4, respectively. (<b>E</b>,<b>F</b>) The ratios of Bcl-2 to β-actin, cleaved-caspase 3 to β-actin expression were normalized, and the values were shown, respectively. Data are shown as the means ± SEM of three biological replicates. Values with different letters (a, b, c) indicate significant differences in bars (<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, comparing the indicated groups.</p> Full article ">
15 pages, 1322 KiB  
Article
Use of Gnawing Hay Blocks: Effects on Productive Performance, Behavior and Reactivity of Growing Rabbits Kept in Parks with Different Sex-Group Compositions
by Marco Birolo, Angela Trocino, Andrea Zuffellato, Fabrizio Pirrone, Francesco Bordignon and Gerolamo Xiccato
Animals 2022, 12(9), 1212; https://doi.org/10.3390/ani12091212 - 8 May 2022
Cited by 1 | Viewed by 2216
Abstract
To improve animal welfare in collective park housing systems, this study assessed the effects of the provision of gnawing hay blocks and the group composition (F: females, M: males, FM: mixed sex) on performance, behavior, and reactivity of 288 growing rabbits reared in [...] Read more.
To improve animal welfare in collective park housing systems, this study assessed the effects of the provision of gnawing hay blocks and the group composition (F: females, M: males, FM: mixed sex) on performance, behavior, and reactivity of 288 growing rabbits reared in 18 parks (16 rabbits/park) from 31 to 73 days of age. The presence of gnawing blocks inside the feeding area of the parks scarcely affected performance and budget time, but decreased the time spent in the resting area compared to parks without blocks (p < 0.001); it increased the time spent moving during the open field test (p < 0.05) and the rate of rabbits that approached the object in the novel-object test (p < 0.05). As for sex-group composition, the feed conversion ratio was lower in parks F and FM than in park M (p < 0.05). During the open-field test, FM rabbits spent more time moving (p < 0.05), whereas M rabbits displayed self-grooming for a longer time (p < 0.01). Results related to production, behavior, and reactivity indicate the provision of gnawing blocks for welfare improvement, but do not support the change from current mixed-sex to single-sex rearing. Full article
(This article belongs to the Special Issue Poultry and Rabbit Production in Alternative Systems)
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<p>Time (% of observation time) spent (<b>a</b>) in the resting area, (<b>b</b>) resting in the resting area, (<b>c</b>) resting in the feeding area, and (<b>d</b>) resting with stretched body in growing rabbits kept in collective parks with only females (F), with only males (M), or with females and males (FM) in parks without (absence) or with (presence) gnawing hay blocks (significant interactions between group composition and presence/absence of gnawing blocks). Data are reported as the means ± standard deviations.</p> Full article ">Figure 2
<p>Time (% of observation time) spent (<b>a</b>) in the resting area, (<b>b</b>) resting in the resting area, (<b>c</b>) and resting in the feeding area of the parks by growing rabbits kept in collective parks without (absence) or with (presence) gnawing hay blocks at 42 and 70 days of age (significant interactions age × absence/presence of gnawing blocks). Data are reported as means ± standard deviations.</p> Full article ">Figure 3
<p>Time (% of observation time) spent (<b>a</b>) sniffing, (<b>b</b>) resting in the feeding area of the park, (<b>c</b>) resting with crouched body, and (<b>d</b>) resting with stretched body in growing rabbits kept in collective parks with only females (F), only males (M) or with males and females (FM) at 42 and 70 days of age (significant interactions between sex-group composition and age). Data are reported as means ± standard deviations.</p> Full article ">
41 pages, 578 KiB  
Review
Alternative Proteins for Fish Diets: Implications beyond Growth
by Cláudia Aragão, Ana Teresa Gonçalves, Benjamín Costas, Rita Azeredo, Maria João Xavier and Sofia Engrola
Animals 2022, 12(9), 1211; https://doi.org/10.3390/ani12091211 - 7 May 2022
Cited by 72 | Viewed by 8700
Abstract
Aquaculture has been challenged to find alternative ingredients to develop innovative feed formulations that foster a sustainable future growth. Given the most recent trends in fish feed formulation on the use of alternative protein sources to decrease the dependency of fishmeal, it is [...] Read more.
Aquaculture has been challenged to find alternative ingredients to develop innovative feed formulations that foster a sustainable future growth. Given the most recent trends in fish feed formulation on the use of alternative protein sources to decrease the dependency of fishmeal, it is fundamental to evaluate the implications of this new paradigm for fish health and welfare. This work intends to comprehensively review the impacts of alternative and novel dietary protein sources on fish gut microbiota and health, stress and immune responses, disease resistance, and antioxidant capacity. The research results indicate that alternative protein sources, such as terrestrial plant proteins, rendered animal by-products, insect meals, micro- and macroalgae, and single cell proteins (e.g., yeasts), may negatively impact gut microbiota and health, thus affecting immune and stress responses. Nevertheless, some of the novel protein sources, such as insects and algae meals, have functional properties and may exert an immunostimulatory activity. Further research on the effects of novel protein sources, beyond growth, is clearly needed. The information gathered here is of utmost importance, in order to develop innovative diets that guarantee the production of healthy fish with high quality standards and optimised welfare conditions, thus contributing to a sustainable growth of the aquaculture industry. Full article
(This article belongs to the Special Issue Advances in Dietary Protein Research: Shaping Innovative Feeds for a Sustainable Ocean)
9 pages, 1042 KiB  
Article
Screening and Carrier Rate of Neuronal Ceroid Lipofuscinosis in Chihuahua Dogs in Japan
by Shahnaj Pervin, Md Shafiqul Islam, Naomi Tada, Toshihiko Tsutsui, Mohammad Mahbubur Rahman, Akira Yabuki, Martia Rani Tacharina, Tofazzal Md Rakib, Shinichiro Maki and Osamu Yamato
Animals 2022, 12(9), 1210; https://doi.org/10.3390/ani12091210 - 7 May 2022
Cited by 3 | Viewed by 2607
Abstract
Neuronal ceroid lipofuscinosis (NCL) is a group of rare lethal neurodegenerative lysosomal storage diseases that occur in a range of dog breeds, including Chihuahuas. Recently, a homozygous single base-pair deletion (c.846delT), which causes a frame shift generating a premature stop codon (p.Phe282Leufs13*) in [...] Read more.
Neuronal ceroid lipofuscinosis (NCL) is a group of rare lethal neurodegenerative lysosomal storage diseases that occur in a range of dog breeds, including Chihuahuas. Recently, a homozygous single base-pair deletion (c.846delT), which causes a frame shift generating a premature stop codon (p.Phe282Leufs13*) in the canine CLN7/MFSD8 gene, has been identified as a causative mutation for NCL in Chihuahuas. The objective of this study was to determine the frequency of the mutant allele and/or carrier rate of NCL in Chihuahuas in Japan using a newly designed real-time PCR assay. Samples of saliva were randomly collected from 1007 Chihuahua puppies during physical examinations prior to the transportation to pet shops. Screening results revealed a carrier rate of 1.29%, indicating a mutant allele frequency (0.00645) that is considered sufficiently high to warrant measures for the control and prevention of this lethal disease. The genotyping assay designed in this study could make a valuable contribution to the control and prevention of NCL. Full article
(This article belongs to the Special Issue Breeding Dogs and Cats: For the Suitability of Kittens, Puppies and Their Breeders)
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<p>Real-time PCR amplification plots of wild-type and mutant alleles for neuronal ceroid lipofuscinosis in Chihuahuas. Amplification was plotted as fluorescence intensity (ΔRn values) against the cycle number. ΔRn values represent the reporter dye signal normalized to the internal reference dye and corrected for the baseline signal established in the initial few PCR cycles. The three amplification plots show the homozygous wild-type (<b>A</b>), heterozygous carrier (<b>B</b>), and homozygous mutant (affected) (<b>C</b>) genotypes. The solid and dotted lines indicate amplification in the presence of wild-type and mutant alleles, respectively.</p> Full article ">Figure 2
<p>An allelic discrimination plot of end point fluorescence real-time PCR data showing the three genotypes (homozygous wild-type, heterozygous carrier, and homozygous mutant) of neuronal ceroid lipofuscinosis in Chihuahuas. This plot presents the data obtained for 45 representative DNA samples of Chihuahuas that had been previously genotyped. The plot shows the fluorescence intensities (ΔRn values) for each allele. ×: No template control; ○: Homozygous wild-type genotype (20 samples); ∆: Heterozygous carrier genotype (13 samples); □: Homozygous mutant (affected) genotype (12 samples).</p> Full article ">
11 pages, 2510 KiB  
Article
Anti-Müllerian Hormone Inhibits FSH-Induced Cumulus Oocyte Complex In Vitro Maturation and Cumulus Expansion in Mice
by Xue Yu, Zan Li, Xinzhe Zhao, Liping Hua, Shuanghang Liu, Changjiu He, Liguo Yang, John S. Davis and Aixin Liang
Animals 2022, 12(9), 1209; https://doi.org/10.3390/ani12091209 - 7 May 2022
Cited by 8 | Viewed by 5828
Abstract
Anti-Müllerian hormone (AMH) is secreted by the ovaries of female animals and exerts its biological effects through the type II receptor (AMHR2). AMH regulates follicular growth by inhibiting the recruitment of primordial follicles and reducing the sensitivity of antral follicles to FSH. Despite [...] Read more.
Anti-Müllerian hormone (AMH) is secreted by the ovaries of female animals and exerts its biological effects through the type II receptor (AMHR2). AMH regulates follicular growth by inhibiting the recruitment of primordial follicles and reducing the sensitivity of antral follicles to FSH. Despite the considerable research on the actions of AMH in granulosa cells, the effect of AMH on the in vitro maturation of oocytes remains largely unknown. In the current study, we showed that AMH is only expressed in cumulus cells, while AMHR2 is produced in both cumulus cells and oocytes. AMH had no significant effect on COCs nuclear maturation, whereas it inhibited the stimulatory effects of FSH on COCs maturation and cumulus expansion. Moreover, AMH treatment effectively inhibited the positive effect of FSH on the mRNA expressions of Hyaluronan synthase 2 (Has2), Pentraxin 3 (Ptx3), and TNF-alpha-induced protein 6 (Tnfaip 6) genes in COCs. In addition, AMH significantly decreased the FSH-stimulated progesterone production, but did not change estradiol levels. Taken together, our results suggest that AMH may inhibit the effects of FSH-induced COCs in vitro maturation and cumulus expansion. These findings increase our knowledge of the functional role of AMH in regulating folliculogenesis. Full article
(This article belongs to the Special Issue Reproductive Biology of Animals: A Focus on Gamete Biology and Role of Antioxidants)
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<p>Expression of <span class="html-italic">Amh</span> and <span class="html-italic">Amhr</span> in mouse cumulus cells and oocytes. PCR amplification product of representative agarose gel electrophoresis. RNA was isolated from cumulus cells (Lane 1), cumulus oocyte complexes (Lane 2), and cumulus-free oocytes (Lane 3). One-microliter amounts of cDNA were used as templates for <span class="html-italic">Amh</span> (<b>A</b>) and <span class="html-italic">Amhr</span> (<b>B</b>) amplification, ddH<sub>2</sub>O was used as negative control for PCR amplification (Lane 4).</p> Full article ">Figure 2
<p>Effect of rhAMH on COCs in vitro maturation. COCs were supplemented with AMH and FSH alone or in combination for 16 h, the first polar body extrusion, mRNA expression in cell lysate, cAMP and MPF levels in the cytoplasm were analyzed. (<b>A</b>), Polar body extrusion percentage; (<b>B</b>–<b>E</b>), the relative mRNA expression of <span class="html-italic">Fshr</span>, <span class="html-italic">Amhr2, Bmp15</span>, and <span class="html-italic">Gdf9</span>; (<b>F</b>), cAMP levels; (<b>G</b>), MPF levels. Data were expressed as mean ± SEM from at least three independent experiments. Bars with different letters represent significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 3
<p>Effect of rhAMH on cumulus cell expansion. COCs were supplemented with AMH and FSH alone or in combination for 16 h, cumulus cell expansion was analyzed by the cumulus expansion index (CEI). (<b>A</b>), representative images of different grades of cumulus cell expansion, a = 0, b = 1, c = 2, d = 3, e = 4, the scale bar represents 50 μm. (<b>B</b>), the representative cumulus cell expansion of each group, the scale bar represents 50 μm. (<b>C</b>), the CEI of each group. Data were from at least three independent experiments. Bars with different letters represent significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 4
<p>Effect of rhAMH on expression of cumulus expansion related genes. COCs were supplemented with AMH and FSH alone or in combination for 16 h,relative expression of transcripts for <span class="html-italic">Ptgs2</span> (<b>A</b>), <span class="html-italic">Has2</span> (<b>B</b>), <span class="html-italic">Ptx3</span> (<b>C</b>), and <span class="html-italic">Tnfaip6</span> (<b>D</b>) genes in cumulus cells were detected by RT-qPCR. The results were evaluated as the relative ratio of the expression level of each mRNA to that of <span class="html-italic">β-actin</span> and were expressed as mean ± SEM from three independent experiments. Bars with different letters represent significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5
<p>Effect of rhAMH treatment on estradiol (<b>A</b>) and progesterone (<b>B</b>) productions in COCs. The data were expressed as mean ± SEM from three independent experiments. Bars with different letters represent significant differences (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">
14 pages, 943 KiB  
Article
Follicular Dynamics during Estrous Cycle of Pubertal, Mature and Postpartum Crossbred (Nili Ravi × Jianghan) Buffaloes
by Adili Abulaiti, Umair Riaz, Zahid Naseer, Zulfiqar Ahmed, Guohua Hua and Liguo Yang
Animals 2022, 12(9), 1208; https://doi.org/10.3390/ani12091208 - 7 May 2022
Cited by 3 | Viewed by 3884
Abstract
The follicular dynamics is used as a reliable indicator for reproductive management in livestock. However, the follicular dynamics (follicle wave emergence, estrus cycle length, diameter of dominant follicle, follicular growth and atretic phases) during the estrous cycle of crossbred (Nili Ravi-Jianghan) buffalo is [...] Read more.
The follicular dynamics is used as a reliable indicator for reproductive management in livestock. However, the follicular dynamics (follicle wave emergence, estrus cycle length, diameter of dominant follicle, follicular growth and atretic phases) during the estrous cycle of crossbred (Nili Ravi-Jianghan) buffalo is still unexplored. Therefore, the present study aimed to observe the follicular dynamics in estrous cycle of crossbred buffaloes at different physiological stages (pubertal; n = 28, sexual mature; n = 22 and postpartum; n = 18). In the present study, the follicular dynamics were ultrasonically examined at 12 h intervals throughout an estrous cycle during the breeding season. The results indicate that about 86.76% (59/68) crossbred buffaloes, irrespective of physiological stage, exhibited two follicular waves in estrous cycle with an average estrus cycle length was 20.7 ± 0.4 days. The estrus cycle length was significantly shorter (p < 0.05) in pubertal buffaloes (19.4 ± 0.4 days) compared with sexual mature (21.5 ± 0.3 days) and postpartum (21.9 ± 0.4 days) buffaloes. The first follicular wave emerged on same day during one- (pubertal vs. postpartum), two- (pubertal vs. mature vs. postpartum) or three-wave (mature vs. postpartum) estrous cycle buffaloes. The maximum diameter of dominant follicle (DF) in pubertal, sexually mature and postpartum crossbred buffaloes was 9.6 ± 2.0 mm, 10.6 ± 0.5 mm and 12.6 ± 0.7 mm with growth rate of 1.08 ± 0.04 mm/day, 0.92 ± 0.04 mm/day, and 0.9 ± 0.07 mm/day, respectively. In conclusion, similar to other buffalo breeds, Nili Ravi-Jianghan crossbred buffaloes showed the two-wave follicular pattern dominantly with an average duration of ~20 days estrous cycle. The observed follicular dynamics can be used as a reliable indicator for synchronization and fixed-time artificial insemination (FTAI) programs to improve the fertility of crossbred buffaloes. Full article
(This article belongs to the Special Issue Recent Developments in Buffalo Reproduction)
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<p>The growth profile of DF in pubertal buffaloes with a one- ((<b>A</b>), <span class="html-italic">n</span> = 3) and two-wave ((<b>B</b>), <span class="html-italic">n</span> = 25) estrous cycle.</p> Full article ">Figure 2
<p>The growth profile of DF in sexually mature crossbred buffaloes with a two- ((<b>A</b>), <span class="html-italic">n</span> = 20) and three-wave ((<b>B</b>), <span class="html-italic">n</span> = 2) estrous cycle.</p> Full article ">Figure 3
<p>The growth profile of DF in postpartum crossbred buffaloes with one- ((<b>A</b>), <span class="html-italic">n</span> = 2), two- ((<b>B</b>), <span class="html-italic">n</span> = 14) and three-wave ((<b>C</b>), <span class="html-italic">n</span> = 2) estrous cycle.</p> Full article ">
18 pages, 1273 KiB  
Article
Is the Hitchcock Story Really True? Public Opinion on Hooded Crows in Cities as Input to Management
by László Kövér, Petra Paládi, Isma Benmazouz, Andrej Šorgo, Natalija Špur, Lajos Juhász, Péter Czine, Péter Balogh and Szabolcs Lengyel
Animals 2022, 12(9), 1207; https://doi.org/10.3390/ani12091207 - 7 May 2022
Cited by 4 | Viewed by 2424
Abstract
In recent years, the Hooded crow (Corvus cornix) has become one of the most successful wild bird species in urban environments across Europe. Hooded crows can cause several problems in cities, including trash scattering, noise disturbance, and aggressive behavior toward humans [...] Read more.
In recent years, the Hooded crow (Corvus cornix) has become one of the most successful wild bird species in urban environments across Europe. Hooded crows can cause several problems in cities, including trash scattering, noise disturbance, and aggressive behavior toward humans or pets, and they can be potential vectors of pathogens. To find effective solutions, the public has to be involved in the decision-making process in urban planning management, managed by the city administration. In this study, we surveyed the attitude of people in Hungary towards crows and crow management by collecting information using an online questionnaire containing 65 questions published in 14 Facebook groups. We found that many people were familiar with corvid species and had personal experience with them. In most cases, these experiences were not negative, so the crows were not or only rarely perceived to cause problems to people, such as aggressive behavior, damage to cars or stealing something. Most respondents recognized that the presence of large numbers of hooded crows is a problem to be solved and acknowledged that they do not know how to resolve it. The majority of people expressed their interest in raising public awareness of crows but not in their management actions, which they believe should be implemented by experts. Most respondents preferred passive, harmless methods. More direct methods such as egg/chick removal from the nest, control by trapping, poisoned baits or firearms, or oral contraceptives were the least acceptable. These results express the difficulty in identifying a control method for managing hooded crow populations that is both acceptable to most people and effective at the same time. This study demonstrates the importance of involving public opinion in wildlife management and providing more information to citizens to reduce human-crow conflicts. Full article
(This article belongs to the Section Birds)
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<p>Locations of local community Facebook user groups in which the questionnaire was published. Source: Google Earth.</p> Full article ">Figure 2
<p>Results of the principal component analysis on responses to statements on coexistence with hooded crows. Note: total explained variance: 55.2%; Bartlett’s test of sphericity: <span class="html-italic">χ</span><sup>2</sup> = 14219.47, <span class="html-italic">p</span> &lt; 0.001; KMO = 0.932; Cronbach’s α = 0.815. OP1: I would like to be involved in projects that aim to raise attention to and awareness of crows. OP2: Colonization of cities by crows is a problem that should be solved. OP3: For the management of crow numbers, all measures by experts are acceptable. OP4: I find measures to control crow populations acceptable and support them. OP5: Damages caused by crows are minor and do not justify population control measures. OP6: The number of crows should be reduced regardless of the type of their habitat. OP7: Crows should not be bothered as their numbers will reach a natural balance. OP8: We should protect crows regardless of the type of their habitat. OP9: I would like to participate in projects aiming to control the population of crows. OP10: The hooded crow is just one of many bird species that should enjoy unlimited protection. OP11: I would sign a petition against measures to control the population of crows. OP12: Damages caused by crows should be reimbursed but should not justify population control. OP13: Crows should be de-listed as game species, which would make their protection easier. OP14: Only the numbers of crows living in cities should be reduced. OP15: City crows should enjoy unlimited legal protection. OP16: Claims to control populations come from the hunting lobby, who aim to shoot more crows. OP17: Crows are wild and do not belong in cities, so they should be removed from urban areas. OP18: Crows should be de-listed as game species, which allows unlimited control measures. OP20: Urban crow numbers should increase as they contribute to the diversity of the urban areas.</p> Full article ">Figure 3
<p>Results of the principal component analysis on responses regarding the acceptability of management methods of hooded crow populations. Note: total explained variance: 51.2%; Bartlett’s test of sphericity: <span class="html-italic">χ</span><sup>2</sup> = 11703.235, <span class="html-italic">p</span> &lt; 0.001; KMO = 0.918; Cronbach α = 0.897. ME1: Scaring crows using techniques that do not cause noise. ME2: All measures for scaring crows. ME3: Scaring crows with noise. ME4: All measures to control the number of adult crows by authorized persons. ME5: All measures to control nesting success. ME6: Use of traps to capture crows in urban areas. ME7: Use of traps on farmland or in the countryside. ME8: Persecution of breeding crows using domesticated birds of prey (falconry). ME9: Physical removal of crows’ nests in urban areas. ME10: Chemical sterilisation of crows. ME11: Shooting adult crows on farmland or in the countryside. ME12: Removal of chicks or eggs from nests. ME13: Shooting adult crows in urban areas. ME14: Crows should be considered as rats, and could be eliminated by anyone. ME15: Shooting crow chicks in nests. ME16: Setting poisoned bait for crows.</p> Full article ">
12 pages, 421 KiB  
Article
Evaluation of the Nutritive Value and Digestibility of Sprouted Barley as Feed for Growing Lambs: In Vivo and In Vitro Studies
by Hani H. Al-Baadani, Abdullah N. Alowaimer, Mohammed A. Al-Badwi, Mutassim M. Abdelrahman, Walid H. Soufan and Ibrahim A. Alhidary
Animals 2022, 12(9), 1206; https://doi.org/10.3390/ani12091206 - 7 May 2022
Cited by 8 | Viewed by 3147
Abstract
The main objective of this study was to investigate the effects of freshly sprouted barley on the growth of lambs, in addition to its nutritional value and digestibility. In addition, sprouted barley digestibility and rumen fermentation were studied in vitro on a dry [...] Read more.
The main objective of this study was to investigate the effects of freshly sprouted barley on the growth of lambs, in addition to its nutritional value and digestibility. In addition, sprouted barley digestibility and rumen fermentation were studied in vitro on a dry matter (DM) basis. A total of 45 three-month-old Awassi lambs were randomly assigned to five treatments of sprouted barley (0, 25, 50, 75, 100%) diets. Bodyweight, weight gain, feed intake and feed efficiency were recorded every two weeks. Nutrient analyses were performed on feed, faecal, and urine samples. DM and non-fibrous carbohydrates were measured. Digestibility of DM, organic matter (OM), and neutral detergent fiber (NDF), as well as gas production, pH value, ammonia-N, and volatile fatty acids (VFAs), were determined in vitro using continuous culture. The results showed that final bodyweight was lower (p < 0.05), while feed intake and the feed-to-gain ratio were increased (p < 0.05) in sprouted barley treatments. Nutrient analysis indicators of sprouted barley treatments (25 to100%) were lower (p < 0.05) for DM, crude protein, acid detergent fiber, lignin and ash, and higher for total digestible nutrients, NDF, fat, phosphorus, zinc, copper, and net energy than the traditional diet. In the in vivo study, the digestibility of nutrients in sprouted barley treatments was improved (p < 0.05), while the diet (sprouted barley 100%) had the lowest digestibility of DM, OM, and NDF compared with the other treatments in the in vitro study. In conclusion, the addition of sprouted barley improved digestibility, and fermentation characteristics, while having a negative effect on growth. Further studies are recommended for optimal growth performance. Full article
(This article belongs to the Collection Feeding Strategies to Improve Sustainability and Welfare in Animal Production)
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<p>Cumulative CO2 gas production during the in vitro digestion for dietary treatments from 2 to 24 h.</p> Full article ">
18 pages, 482 KiB  
Article
Sex Differences in Mate Choice Preference Characteristics of Aequidens rivulatus
by Haixia Li, Jie Wang, Xu Zhang, Yu Hu, Qinglin Cai, Ying Liu and Zhen Ma
Animals 2022, 12(9), 1205; https://doi.org/10.3390/ani12091205 - 7 May 2022
Cited by 2 | Viewed by 16968
Abstract
The mating roles of males and females, to a certain extent, are dynamic and variable. Several factors influence the mate choice process. Nonetheless, the main preference features have not yet been fully understood in Aequidens rivulatus. In this study, because of its [...] Read more.
The mating roles of males and females, to a certain extent, are dynamic and variable. Several factors influence the mate choice process. Nonetheless, the main preference features have not yet been fully understood in Aequidens rivulatus. In this study, because of its natural pairing characteristics, A. rivulatus was selected to explore the mate choice preferences of different sexes. Specifically, male and female behavioral performances were described and quantified through a “no-choice paradigm” during mate choice. A total of 12 behavioral performances were defined in male mate choice (experiment 1), whereas 14 behavioral performances were defined in female mate choice (experiment 2). According to the obtained results, unselected females did not display any proactive behaviors in experiment 1, whereas unselected males exhibited proactive behaviors in experiment 2, including quivering, nipping, tail beating, swimming up and down, and aggression. It was also found that both male and female individuals tend to express dislike rather than like. Those behaviors with higher frequencies (e.g., quivering) often mean less energy expenditure, thus easier repeatability. Moreover, principal component analysis (PCA) was employed to extract and identify mate choice preference features. Preliminary results indicated that male preferences for a mate were mainly associated with body size, behavioral intention, and appearance, whereas the intensity of female preferences was in the order of body size, appearance, and behavioral intention. In addition, sex hormone levels were associated with mate choices. Full article
(This article belongs to the Collection Behavioral Ecology of Aquatic Animals)
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<p>The behavioral intention of male and female fish in Experiment 1. (<b>a</b>) Frequency of male’s behavioral performance; (<b>b</b>) Proportion of duration of behavioral performance in males; (<b>c</b>) Frequency of female’s behavior performance; (<b>d</b>) Proportion of the duration of behavior performance in females.</p> Full article ">Figure 2
<p>Body size preference for the different sexes. (<b>a</b>) Female body size preference in Experiment 1; (<b>b</b>) Male body size preference in Experiment 2. A significant difference is denoted by an asterisk.</p> Full article ">Figure 3
<p>The ornamental preference for the different sexes. (<b>a</b>) Male ornamental preference in Experiment 1; (<b>b</b>) Female ornamental preference in Experiment 2. A significant difference is denoted by an asterisk.</p> Full article ">Figure 4
<p>Comparison of sex hormone levels. (<b>a</b>) Estradiol levels in females in Experiment 1; (<b>b</b>) Testosterone levels in males in Experiment 2. A significant difference is denoted by an asterisk.</p> Full article ">Figure 5
<p>The behavioral intention of male and female fish in Experiment 2. (<b>a</b>) Frequency of female’s behavioral performance; (<b>b</b>) Proportion of the duration of behavioral performance in females; (<b>c</b>) Frequency of male’s behavior performance; (<b>d</b>) Proportion of the duration of behavior performance in males.</p> Full article ">
13 pages, 2768 KiB  
Article
Transcriptome Revealed Exposure to the Environmental Ammonia Induced Oxidative Stress and Inflammatory Injury in Spleen of Fattening Pigs
by Yongjie Chen, Runxiang Zhang, Susu Ding, Haoyang Nian, Xiangyin Zeng, Honggui Liu, Houjuan Xing, Jianhong Li, Jun Bao and Xiang Li
Animals 2022, 12(9), 1204; https://doi.org/10.3390/ani12091204 - 7 May 2022
Cited by 5 | Viewed by 2407
Abstract
Ammonia is one of the major environmental pollutants that seriously threaten human health. Although many studies have shown that ammonia causes oxidative stress and inflammation in spleen tissue, the mechanism of action is still unclear. In this study, the ammonia poisoning model of [...] Read more.
Ammonia is one of the major environmental pollutants that seriously threaten human health. Although many studies have shown that ammonia causes oxidative stress and inflammation in spleen tissue, the mechanism of action is still unclear. In this study, the ammonia poisoning model of fattening pigs was successfully established. We examined the morphological changes and antioxidant functions of fattening pig spleen after 30-day exposure to ammonia. Effects of ammonia in the fattening pig spleen were analyzed from the perspective of oxidative stress, inflammation, and histone methylation via transcriptome sequencing technology (RNA-seq) and real-time quantitative PCR validation (qRT-PCR). We obtained 340 differential expression genes (DEGs) by RNA-seq. Compared with the control group, 244 genes were significantly upregulated, and 96 genes were significantly downregulated in the ammonia gas group. Some genes in Gene Ontology (GO) terms were verified and showed significant differences by qRT-PCR. The KEGG pathway revealed significant changes in the MAPK signaling pathway, which is strongly associated with inflammatory injury. To sum up, the results indicated that ammonia induces oxidative stress in pig spleen, activates the MAPK signaling pathway, and causes spleen necrosis and injury. In addition, some differential genes encoding epigenetic factors were found, which may be involved in the response mechanism of spleen tissue oxidative damage. The present study provides a transcriptome database of ammonia-induced spleen poisoning, providing a reference for risk assessment and comparative medicine of ammonia. Full article
(This article belongs to the Section Pigs)
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<p>Histological structure of the spleens stained by hematoxylin and eosin (×20) (<span class="html-italic">n</span> = 3/group): (<b>A</b>) control group; (<b>B</b>) ammonia group. The histopathological lesions included the volume of splenic nodules decreased. Lymphocyte count (black arrow) was reduced.</p> Full article ">Figure 2
<p>Changes in oxidative stress indicators induced by ammonia exposure (GSH-Px activity, SOD activity, GSH activity, and MDA content) in pig spleen. Data are expressed as mean ± standard deviation. * indicates a significant difference (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 3
<p>(<b>A</b>) Heat map of DEGs. The abscissa represents the sample, and the ordinate represents the gene. Different colors indicate different levels of gene expression. Red represents highly expressed genes, and dark blue represents low expressed genes. (<b>B</b>) Volcano plots of DEGs. The x-coordinate is log2 (fold-change), and the y-coordinate is −log10 (<span class="html-italic">p</span>-value). Red and blue dots represent upregulated and downregulated DEGs, respectively. (<b>C</b>) Column diagram of DEGs. The red column represents the number of upregulated genes, and the blue column represents the number of downregulated genes.</p> Full article ">Figure 4
<p>Top 20 pathways showed significant changes in histone methylation-related genes. MAPK signaling pathway was highlighted in red circles.</p> Full article ">Figure 5
<p>(<b>A</b>) GO enrichment analysis of DEGs. (<b>B</b>) Top 20 GO terms in GO enrichment analysis. Rich factor represents the number of DEGs enriched in the GO terms.</p> Full article ">Figure 6
<p>Forecast tendency of histone methyltransferase genes, oxidative stress, inflammatory, and MAPK signaling pathways in the pig spleen (<span class="html-italic">n</span> = 6/group): (<b>A</b>) histone methyltransferase genes; (<b>B</b>) oxidative stress; (<b>C</b>) inflammatory; (<b>D</b>) MAPK signaling pathways. (<b>E</b>) Comparison and analysis of RNA-Seq and qRT-PCR results.</p> Full article ">
17 pages, 3683 KiB  
Article
Immune Functions Alterations Due to Racing Stress in Thoroughbred Horses
by Mônica Cristina Zandoná Meleiro, Hianka Jasmyne Costa de Carvalho, Rafaela Rodrigues Ribeiro, Mônica Duarte da Silva, Cristina Massoco Salles Gomes, Maria Angélica Miglino and Irvênia Luiza de Santis Prada
Animals 2022, 12(9), 1203; https://doi.org/10.3390/ani12091203 - 7 May 2022
Cited by 3 | Viewed by 2338
Abstract
Racehorses are constantly exposed to stress. Aiming to verify the state of blood components and cortisol alterations during their routine and after races, phagocytosis and oxidative neutrophil burst assays, serum cortisol determination, erythrocytes apoptosis evaluation, lymphoproliferation assays, and blood count tests were performed [...] Read more.
Racehorses are constantly exposed to stress. Aiming to verify the state of blood components and cortisol alterations during their routine and after races, phagocytosis and oxidative neutrophil burst assays, serum cortisol determination, erythrocytes apoptosis evaluation, lymphoproliferation assays, and blood count tests were performed in thirty Thoroughbred racehorses, which were divided in two groups. The samples were taken right after races (moment 0 d), during rest periods (−11 d, +1 d, +3 d), and after training (−8, +2, +5). In both groups, the phagocytosis showed a decrease in percentage and intensity immediately after the race when comparing samples collected during rest or training periods. In the mean values of oxidative burst on samples collected immediately after the race, group I animals demonstrated a decrease (524.2 ± 248.9) when compared with those samples collected in other moments. No significant differences were found between the results of different moments regarding the apoptotic cells and lymphoproliferation assays. The mean values of serum cortisol levels were increased immediately after racing. There was an increase in the percentage of neutrophils found immediately after the race. It was possible to conclude that, although a transient reduction was found in the number of neutrophils, the horses’ adaptive function was not affected. Full article
(This article belongs to the Special Issue Hormones and the Welfare of Animals)
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<p>Representation of peripheral blood leukocyte flow cytometry data of the studied horses, after lysis of erythrocytes, in a dot plot. Cell populations were selected according to their forward scatter (FSC) versus side scatter (SSC) profiles indicating cell size versus granularity and complexity. R1 corresponds to the neutrophil gate, R2 corresponds to lymphocytes, and R3 corresponds to monocytes.</p> Full article ">Figure 2
<p>(<b>A</b>,<b>B</b>): % of phagocytosis performed by neutrophils from animals of group I (<b>A</b>) and group II (<b>B</b>) samples. (<b>C</b>,<b>D</b>): phagocytosis intensity expressed as mean fluorescence intensity (geometric mean of the number of bacteria destroyed by neutrophils) from animals in group I (<b>C</b>) and group II (<b>D</b>). (<b>E</b>,<b>F</b>): mean fluorescence on a logarithmic scale of the neutrophils in the samples of animals of group I (<b>E</b>) and group II (<b>F</b>). Group I—moments −11 d, 0 d, +1 d, +2 d and +3 d. Group II—moments −8 d, 0 d, +1 d and +5 d. a,b = significant statics difference.</p> Full article ">Figure 3
<p>Mean values of total apoptotic cells in moments −8 d, 0 d, +1 d, and +5 d, in the samples of the animals of group II. Statistical difference according to the Tukey–Kramer test—Beltsville, 2003.</p> Full article ">Figure 4
<p>Flow cytometry data, in dot plot, of the peripheral blood mononuclear leukocytes of the studied horses, after a 72-h culture. Cell populations identified according to their forward scatter (FSC) versus side scatter (SSC) profiles, cell size versus granularity and complexity. R4—lymphocyte gate.</p> Full article ">Figure 5
<p>CFSE fluorescence histogram (carboxyfluorescein diacetate) of equine peripheral blood lymphocytes after culture for 72 h at 5% CO<sub>2</sub> and 37 °C without use of mitogens (control). M1 and M2 represent the markers used to enumerate the events of each division cycle that the software used for calculating the proliferation index.</p> Full article ">Figure 6
<p>CFSE fluorescence histogram (carboxyfluorescein diacetate) of lymphocytes from equine peripheral blood after 72 h culture at 5% CO<sub>2</sub> and 37 °C without concavalin A. M1 and M2 represent the markers used to list the events of each division cycle that the software used for calculating the proliferation index.</p> Full article ">Figure 7
<p>Representation of the average values referring to the counting of the relative number (%) of neutrophils (No) at the different collection days defined as −8 d, 0 d, +1 d, and +5 d. Statistical difference according to the Tukey–Kramer test—Beltsville, 2003. a = significant statics difference.</p> Full article ">Figure 8
<p>Representation of the average values of the relative number (%) of lymphocytes (Lo) on the different collection days defined as −8 d, 0 d, +1 d, and +5 d. Statistical difference performed according to the Tukey–Kramer test—Beltsville, 2003. b = significant statics difference.</p> Full article ">Figure 9
<p>Representation of the mean values of serum cortisol levels from the samples of the animals in group I (1 to 15), taken at the different collection moments at −11 d, 0 d, +1 d, +2 d, and +3 d. Statistical difference according to the Tukey–Kramer test—Beltsville, 2003. a = significant statics difference.</p> Full article ">Figure 10
<p>Representation of the mean values of serum cortisol levels of samples from animals in group II (16 to 30), found at the different moments of collection defined as −8 d, 0 d, +1 d, and +5 d. Statistical difference according to the Tukey–Kramer test—Beltsville, 2003. a = significant statics difference.</p> Full article ">
23 pages, 3058 KiB  
Article
Associations between Milk Fatty Acid Profile and Body Condition Score, Ultrasound Hepatic Measurements and Blood Metabolites in Holstein Cows
by Diana Giannuzzi, Alessandro Toscano, Sara Pegolo, Luigi Gallo, Franco Tagliapietra, Marcello Mele, Andrea Minuti, Erminio Trevisi, Paolo Ajmone Marsan, Stefano Schiavon and Alessio Cecchinato
Animals 2022, 12(9), 1202; https://doi.org/10.3390/ani12091202 - 6 May 2022
Cited by 12 | Viewed by 2691
Abstract
Dairy cows have high incidences of metabolic disturbances, which often lead to disease, having a subsequent significant impact on productivity and reproductive performance. As the milk fatty acid (FA) profile represents a fingerprint of the cow’s nutritional and metabolic status, it could be [...] Read more.
Dairy cows have high incidences of metabolic disturbances, which often lead to disease, having a subsequent significant impact on productivity and reproductive performance. As the milk fatty acid (FA) profile represents a fingerprint of the cow’s nutritional and metabolic status, it could be a suitable indicator of metabolic status at the cow level. In this study, we obtained milk FA profile and a set of metabolic indicators (body condition score, ultrasound liver measurements, and 29 hematochemical parameters) from 297 Holstein–Friesian cows. First, we applied a multivariate factor analysis to detect latent structure among the milk FAs. We then explored the associations between these new synthetic variables and the morphometric, ultrasonographic and hematic indicators of immune and metabolic status. Significant associations were exhibited by the odd-chain FAs, which were inversely associated with β-hydroxybutyrate and ceruloplasmin, and positively associated with glucose, albumin, and γ-glutamyl transferase. Short-chain FAs were inversely related to predicted triacylglycerol liver content. Rumen biohydrogenation intermediates were associated with glucose, cholesterol, and albumin. These results offer new insights into the potential use of milk FAs as indicators of variations in energy and nutritional metabolism in early lactating dairy cows. Full article
(This article belongs to the Special Issue Solutions for Breeding Healthy and Efficient Animals: Novel Phenotypes and Omics Approaches for Improving Yield and Quality of Animal Products)
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<p>Least square means (LSM) for F1-de novo synthesis/preformed (<b>a</b>,<b>b</b>) and F2-branched chain fatty acids (BCFA; (<b>c</b>–<b>f</b>)) across the traits of concern. (<b>a,e</b>): advanced oxidation protein products (AOPP); (<b>b</b>,<b>f</b>): ceruloplasmin; c: predicted liver triacylglycerol (pTAG); (<b>d</b>): cholesterol. Black dots indicate the LSM and error bars indicate the standard error. Trendlines define linear or quadratic pattern according to polynomial contrast calculations (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 2
<p>Least square means (LSM) for F3-long chain—polyunsaturated fatty acids (LC-PUFA; (<b>a</b>–<b>d</b>) and F4-alternative rumen biohydrogenation of α-linolenic acid (alternative RBH ALA) (<b>e</b>–<b>h</b>) across the traits of concern. (<b>a</b>): body condition score (BCS); (<b>b</b>,<b>h</b>): alkaline phosphatase (ALP); (<b>c</b>): total thiol groups (SHp); (<b>d</b>): ceruloplasmin; (<b>e</b>): predicted liver triacylglycerol (pTAG); (<b>f</b>): non-esterified fatty acids (NEFA); (<b>g</b>): glucose. Black dots indicate the LSM and error bars indicate the standard error. Trendlines define linear or quadratic pattern according to polynomial contrast calculations (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 3
<p>Least square means (LSM) for F5-production (<b>a</b>,<b>b</b>,<b>c</b>) and F6-short chain fatty acids (SCFA; (<b>d</b>)) across the traits of concern. (<b>a</b>,<b>d</b>): predicted liver triacylglycerol (pTAG); (<b>b</b>): cholesterol; (<b>c</b>): albumin. Black dots indicate the LSM and error bars indicate the standard error. Trendlines define linear or quadratic pattern according to polynomial contrast calculations (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 4
<p>Least square means (LSM) for F7-odd chain fatty acids (OCFA) across the traits of concern. (<b>a</b>): = γ-glutamyl transferase (GGT); (<b>b</b>): total thiol groups (SHp); (<b>c</b>): albumin; (<b>d</b>): glucose; (<b>e</b>): β-hydroxybutyrate acid (BHB); (<b>f</b>): ceruloplasmin; (<b>g</b>): globulins; (<b>h</b>): total proteins. Black dots indicate the LSM and error bars indicate the standard error. Trendlines define linear or quadratic pattern according to polynomial contrast calculations (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">Figure 5
<p>Least square means (LSM) for F8-rumen biohydrogenation (RBH) across the traits of concern. (<b>a</b>): glucose; (<b>b</b>): cholesterol; (<b>c</b>): albumin; (<b>d</b>): advanced oxidation protein products (AOPP). Black dots indicate the LSM and error bars indicate the standard error. Trendlines define linear or quadratic pattern according to polynomial contrast calculations (<span class="html-italic">p</span> &lt; 0.05).</p> Full article ">
17 pages, 2768 KiB  
Article
Supplementing a Phytogenic Feed Additive Modulates the Risk of Subacute Rumen Acidosis, Rumen Fermentation and Systemic Inflammation in Cattle Fed Acidogenic Diets
by Raul Rivera-Chacon, Ezequias Castillo-Lopez, Sara Ricci, Renee M. Petri, Nicole Reisinger and Qendrim Zebeli
Animals 2022, 12(9), 1201; https://doi.org/10.3390/ani12091201 - 6 May 2022
Cited by 18 | Viewed by 3219
Abstract
Feeding with high-concentrate diets increases the risk of subacute ruminal acidosis (SARA). This experiment was conducted to evaluate whether supplementing a phytogenic feed additive based on L-menthol, thymol, eugenol, mint oil (Mentha arvensis) and cloves powder (Syzygium aromaticum) (PHY) [...] Read more.
Feeding with high-concentrate diets increases the risk of subacute ruminal acidosis (SARA). This experiment was conducted to evaluate whether supplementing a phytogenic feed additive based on L-menthol, thymol, eugenol, mint oil (Mentha arvensis) and cloves powder (Syzygium aromaticum) (PHY) can amend the ruminal fermentation profile, modulate the risk of SARA and reduce inflammation in cattle. The experiment was designed as a crossover design with nine non-lactating Holstein cows, and was conducted in two experimental runs. In each run, cows were fed a 100% forage diet one week (wk 0), and were then transitioned stepwise over one week (0 to 65% concentrate, wk adapt.) to a high concentrate diet that was fed for 4 weeks. Animals were fed diets either with PHY or without (CON). The PHY group had an increased ruminal pH compared to CON, reduced time to pH < 5.8 in wk 3, which tended to decrease further in wk 4, reduced the ruminal concentration of D-lactate, and tended to decrease total lactate (wk 3). In wk 2, PHY increased acetate, butyrate, isobutyrate, isovalerate, and the acetate to propionate ratio compared to CON. Phytogenic supplementation reduced inflammation compared to CON in wk 3. Overall, PHY had beneficial effects on ruminal fermentation, reduced inflammation, and modulated the risk of SARA starting from wk 3 of supplementation. Full article
(This article belongs to the Special Issue Advances in Animal Novel Alternative Feed)
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<p>Diurnal variation of ruminal pH in various weeks in cows fed either forage-only or with 65% concentrate in a diet, without supplementation (CON) or supplemented (PHY) with a phytogenic feed additive based on L-menthol, thymol, eugenol, mint oil (<span class="html-italic">Mentha arvensis</span>) and cloves powder (<span class="html-italic">Syzygium aromaticum</span>). Time of feeding: 08:00 h (<span class="html-fig-inline" id="animals-12-01201-i001"> <img alt="Animals 12 01201 i001" src="/animals/animals-12-01201/article_deploy/html/images/animals-12-01201-i001.png"/></span>). (<b>A</b>) Week 0 forage, SEM = 0.05. (<b>B</b>) Day 4 of adaptation week, 40% concentrate, SEM = 0.08. (<b>C</b>) Week 1 of high concentrate, SEM = 0.09. (<b>D</b>) Week 2 of high concentrate, SEM = 0.12. (<b>E</b>) Week 3 of high concentrate, SEM = 0.12. (<b>F</b>) Week 4 of high concentrate, SEM = 0.12. <span class="html-italic">p</span>-Values: Time &lt; 0.01; Week &lt; 0.01; Time × Week &lt; 0.01. SEM: The largest standard error of the mean.</p> Full article ">Figure 2
<p>Variation of ruminal short chain fatty acid fermentation from 0 to 12 h post-feeding in cows fed either forage-only (F) or a high concentrate (HC), without supplementation (CON) or supplemented (PHY) with a phytogenic feed additive based on L-menthol, thymol, eugenol, mint oil (<span class="html-italic">Mentha arvensis</span>) and cloves powder (<span class="html-italic">Syzygium aromaticum</span>). <span class="html-italic">p</span>-Values: Acetate, Time &lt; 0.01, Trt = 0.26, Diet &lt; 0.01, Time × Trt × Diet × Week &lt; 0.01; Propionate, Time &lt; 0.01, Trt &lt; 0.05, Diet &lt; 0.01, Time × Trt × Diet × Week &lt; 0.01; Butyrate, Time &lt; 0.01, Trt = 0.23, Diet &lt; 0.01, Time × Trt × Diet × Week &lt; 0.01; Isobutyrate, Time = 0.36, Trt = 0.28, Diet &lt; 0.01, Time × Trt × Diet × Week &lt; 0.01; Valerate, Time &lt; 0.01, Trt = 0.30, Diet &lt; 0.01, Time × Trt × Diet × Week &lt; 0.01; Isovalerate, Time = 0.45, Trt &lt; 0.05, Diet &lt; 0.01, Time × Trt × Diet × Week &lt; 0.01.</p> Full article ">Figure 3
<p>Variation of ruminal D-lactate, L-lactate (mM), and total ammonia concentration (mg/dL) from 0 to 12 h post-feeding in cows fed either all-forage (F) or a high concentrate (HC), without supplementation (CON) or supplemented (PHY) with a phytogenic feed additive based on L-menthol, thymol, eugenol, mint oil (<span class="html-italic">Mentha arvensis</span>) and cloves powder (<span class="html-italic">Syzygium aromaticum</span>). <span class="html-italic">p</span>-Values: D-lactate, Time = 0.99, Trt = 0.42, Diet &lt; 0.01, Time × Trt × Diet × Week &lt; 0.01; L-lactate, Time = 0.63, Trt = 0.60, Diet &lt; 0.01, Time × Trt × Diet × Week &lt; 0.05; Total ammonia, Time &lt; 0.01, Trt = 0.90, Diet &lt; 0.01, Time × Trt × Diet × Week &lt; 0.01.</p> Full article ">
12 pages, 470 KiB  
Article
Effect of Dietary Organic and Inorganic Sulfur on the Performance of Coccidiosis Vaccine Challenged Broiler Chickens
by Yong-Sung Jeon, Yoo-Bhin Kim, Hyun-Gwan Lee, Jina Park, Yun-Ji Heo, Gyo-Moon Chu and Kyung-Woo Lee
Animals 2022, 12(9), 1200; https://doi.org/10.3390/ani12091200 - 6 May 2022
Cited by 5 | Viewed by 2483
Abstract
The objective of this study was to evaluate the effects of dietary sulfur from either organic (methyl sulfonyl methane, MSM) or inorganic (sodium sulfate, SS) sources on the growth performance of broiler chickens challenged against a high-dose coccidiosis vaccine. A total of 320 [...] Read more.
The objective of this study was to evaluate the effects of dietary sulfur from either organic (methyl sulfonyl methane, MSM) or inorganic (sodium sulfate, SS) sources on the growth performance of broiler chickens challenged against a high-dose coccidiosis vaccine. A total of 320 day-old Ross 308 broiler chicks were randomly placed into 32 pens of 10 birds each (keeping 16 pens/control group and 8 pens/treatment group until 21 days post-hatch) and reared for 28 days. The experimental diets were formulated by mixing a corn and soybean meal-based control diet with MSM or SS. At 21 days post-hatch, half (n = 8) of the control and all of the sulfur-added diet-fed (i.e., MSM and SS) groups were challenged with a 30-fold dose of a commercially available Eimeria vaccine (Livacox® T coccidiosis vaccine). Unchallenged control chicks (n = 8) were considered as the negative control group. At 21 days (before coccidiosis vaccine challenge), the production parameters and cecal short-chain fatty acids were not affected by dietary treatments. The concentrations of total antioxidant capacity in liver samples were elevated in both the MSM and SS groups compared with the control group (p = 0.001). During 21 to 28 days (i.e., one week post coccidiosis vaccine challenge), challenge tended to lower body weight and feed intake by an average of 5.3% (p = 0.262) and 2.8% (p = 0.504), respectively, but to increase the feed conversion ratio by an average of 2.7% (p = 0.087) compared with the non-challenged control groups. None of dietary sulfur groups affected the body weight gain, feed intake, or feed conversion ratio of vaccine-challenged chickens. Mild Eimeria-specific lesions were noted in duodenum (p = 0.006), jejunum (p = 0.017), and ceca (p = 0.047), but dietary sulfur treatments did not affect the Eimeria-induced gut lesion scores. At 28 days, Eimeria challenge significantly impaired (p = 0.001) the apparent ileal digestibility of crude protein and crude ash compared with the naïve control group. Dietary MSM increased the apparent ileal digestibility of crude ash by 15.5% on average compared with the coccidiosis vaccine control group. We conclude that dietary antioxidant sulfur of organic or inorganic origins at the inclusion level (i.e., 0.7 g sulfur/kg of diet) has a limited effect on the growth performance of chickens challenged with coccidiosis vaccine. Full article
(This article belongs to the Section Poultry)
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<p>Schematic outline of the experimental design. Chickens were fed diets containing none, methyl sulfonyl methane, or sodium sulfate until the end of the experiment. Blood, liver, and intestine samples were obtained at 21 days, and all chickens except for the non-challenged control groups were orally challenged with coccidiosis vaccine at 21 days. Intestinal samples were obtained for scoring gut lesions at 27 days (i.e., 6 days of post vaccine challenge). At 28 days, ileal digesta were sampled for assessing ileal nutrient digestibility.</p> Full article ">
15 pages, 952 KiB  
Article
Response of Rambouillet Lambs to an Artificial Gastrointestinal Nematode Infection
by Jacob W. Thorne, Scott A. Bowdridge, Brenda M. Murdoch and R. Reid Redden
Animals 2022, 12(9), 1199; https://doi.org/10.3390/ani12091199 - 6 May 2022
Cited by 6 | Viewed by 1861
Abstract
Gastrointestinal nematodes (GIN) threaten the productivity and health of sheep worldwide, prompting the need for genetic selection to reduce GIN susceptibility. Fecal egg count (FEC), packed-cell volume (PCV), and various production traits were examined in parasitized Rambouillet sheep and compared to sire FEC [...] Read more.
Gastrointestinal nematodes (GIN) threaten the productivity and health of sheep worldwide, prompting the need for genetic selection to reduce GIN susceptibility. Fecal egg count (FEC), packed-cell volume (PCV), and various production traits were examined in parasitized Rambouillet sheep and compared to sire FEC estimated breeding value (EBV). Rambouillet lambs (n = 77) were inoculated with 10,000 H. contortus L3 larvae. Subsequently, FEC, PCV, and body weight (BW) were captured at seven-day intervals for six weeks. Lambs were sired by one of two rams with post-weaning FEC EBV of −9% or +9%. Mean FEC differed (p = 0.0132) with lambs from the lower EBV sire (“Sire L”) being reduced, versus those from the higher EBV sire (“Sire H”), being 2135 ± 211 vs. 2912 ± 207 eggs per gram, respectively. Males and females did not differ for FEC, but females exhibited a higher mean PCV than males, (33.74 vs. 29.65%, p < 0.0001). Lambs were shorn ~120 d post artificial infection and wool measurements were captured. A negative correlation between FEC and grease fleece weight was observed. Our results describe the response of Rambouillet lambs to artificial H. contortus infection and suggest FEC EBV can reduce susceptibility to GIN in this breed. Full article
(This article belongs to the Special Issue Parasites and Parasitic Diseases)
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<p>Response of Rambouillet lambs to an inoculation of 10,000 <span class="html-italic">Haemonchus contortus</span> L3 larvae. Indicators of parasite infection are displayed and include (<b>a</b>) mean fecal egg count and (<b>b</b>) mean packed-cell volume percentage for all lambs on the trial.</p> Full article ">Figure 2
<p>Effects of a <span class="html-italic">H contortus</span> infection on Rambouillet lambs. (<b>a</b>) Mean fecal egg count (FEC) for all lambs, and for each sire group, for weeks three, four, five, and six. At weeks one and two, FEC were zero. <span class="html-italic">p</span>-values for within-week sire comparisons for weeks three, four, five, and six were 0.0082, 0.0094, 0.0503, and 0.2160, respectively. (<b>b</b>) Mean packed-cell volume % (PCV) for all lambs, and for each sire group, across the trial.</p> Full article ">Figure 3
<p>Differing responses to <span class="html-italic">H. contortus</span> infection between female and male Rambouillet lambs. (<b>a</b>) Fecal egg count by sex. Female lambs tended to have higher FEC at week six (<span class="html-italic">p</span> = 0.0562). (<b>b</b>) Mean packed-cell volume % (PCV) for lambs by sex. Female lambs appeared to be more “resilient” to H. contortus infection than males, as evidenced by the greater reduction in PCV of male lambs. For all lambs, <span class="html-italic">H. contortus</span> larvae had begun feeding on host blood by week two, however FEC were not above zero until week three, confirming that larvae begin consuming host blood prior to sexual maturity.</p> Full article ">Figure 4
<p>Average lamb body weight (BW), left axis, and average daily gain (ADG), right axis, across the trial of lambs artificially infected with <span class="html-italic">H. contortus</span>. Lambs were weighed at weekly intervals and ADG was calculated by subtracting the lamb’s BW at the previous week from the BW of the current week, then dividing by the number of days in between those two measurements.</p> Full article ">
17 pages, 332 KiB  
Article
Olive Pomace and Soybean-Sunflower Acid Oils as Alternative Fat Sources in European Seabass (Dicentrarchus labrax) Diets: Effects on Performance, Digestibility and Flesh Fatty Acid Composition and Quality Parameters
by Gerard Verge-Mèrida, Ana Cristina Barroeta, Carlos Ferrer, Tània Serrano, Francesc Guardiola, María Dolores Soler and Roser Sala
Animals 2022, 12(9), 1198; https://doi.org/10.3390/ani12091198 - 6 May 2022
Cited by 5 | Viewed by 2534
Abstract
The effects of dietary inclusion of soybean-sunflower and olive pomace acid oils on growth, digestibility and flesh composition were studied in European seabass. Eight diets were fed for 100 days (101.37 ± 0.33 g initial weight, mean ± SD), differing in the added [...] Read more.
The effects of dietary inclusion of soybean-sunflower and olive pomace acid oils on growth, digestibility and flesh composition were studied in European seabass. Eight diets were fed for 100 days (101.37 ± 0.33 g initial weight, mean ± SD), differing in the added fat source (25% fish oil, 75% experimental oil): S (crude soybean oil), SA (soybean-sunflower acid oil), O (crude olive pomace oil) or OA (olive pomace acid oil); 3 blends: S-O, S-OA, SA-OA at a 1:1 ratio; and a diet containing only fish oil (F) as a control. Animals fed OA showed the worst performance among dietary treatments, with the lowest weight, specific growth ratio, average daily gain and the highest feed conversion ratio (p < 0.01). In contrast, other diets including acid oils did not impair performance. Acid oil diets did not affect the apparent digestibility of dry matter, crude protein or total fatty acids (p > 0.05), but a lower digestibility of lipids and saturated fatty acids was observed (p < 0.001). Flesh composition and fatty acid profile were not affected by the high dietary free FA content (p > 0.05). Hence the results suggest that the studied acid oils may potentially be used in fish diets although further studies are needed. Full article
(This article belongs to the Special Issue Advances in Animal Novel Alternative Feed)
15 pages, 1808 KiB  
Article
Establishment and Application of an Indirect Enzyme-Linked Immunosorbent Assay for Measuring GPI-Anchored Protein 52 (P52) Antibodies in Babesia gibsoni-Infected Dogs
by Qin Liu, Xueyan Zhan, Dongfang Li, Junlong Zhao, Haiyong Wei, Heba Alzan and Lan He
Animals 2022, 12(9), 1197; https://doi.org/10.3390/ani12091197 - 6 May 2022
Cited by 1 | Viewed by 2114
Abstract
Babesia gibsoni is a malaria-like protozoan that parasitizes the red blood cells of canids to cause babesiosis. Due to its high expression and essential function in the survival of parasites, the Glycosylphosphatidylinositol (GPI) anchor protein family is considered an excellent immunodiagnostic marker. Herein, [...] Read more.
Babesia gibsoni is a malaria-like protozoan that parasitizes the red blood cells of canids to cause babesiosis. Due to its high expression and essential function in the survival of parasites, the Glycosylphosphatidylinositol (GPI) anchor protein family is considered an excellent immunodiagnostic marker. Herein, we identified a novel GPI-anchored protein named as BgGPI52-WH with a size of 52 kDa; the recombinant BgGPI52-WH with high antigenicity and immunogenicity was used as a diagnostic antigen to establish a new iELISA method. The iELISA had a sensitivity of 1:400, and no cross-reaction with other apicomplexan parasites occurred. We further demonstrated that the degree of variation was less than 10% using the same samples from the same or different batches of an enzyme-labeled strip. It was found that the method was able to detect early infection (6 days after infection) in the sera of the B. gibsoni-infected experimental dogs in which antibody response to rBgGPI52-WH was evaluated. Clinical sera from pet hospitals were further tested, and the average positive rate was about 11.41% (17/149). The results indicate that BgGPI52-WH is a reliable diagnostic antigen, and the new iELISA could be used as a practical method for the early diagnosis of B. gibsoni. Full article
(This article belongs to the Collection Advances in Parasite Epidemiology and Population Genetics)
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<p>Sequence analysis of BgGPI52-WH. (<b>a</b>) Identification of target gene sequences. Lane M—marker; lane 1—the BgGPI52-WH target band amplified from <span class="html-italic">B. gibsoni</span> gDNA; lane 2—the BgGPI52-WH target band amplified from <span class="html-italic">B. gibsoni</span> cDNA; lane 3—control group. (<b>b</b>) A schematic diagram showing the TM domain, signal peptide (SP) and GPI anchor site of the target protein sequence. (<b>c</b>) Prediction of the antigen epitopes of the BgGPI52-WH protein. Purple represents the antigenic index; the higher the antigenic index, the better the antigenicity. Yellow represents the probability index of protein on the surface.</p> Full article ">Figure 2
<p>Prokaryotic expression of recombinant protein BgGPI52-WH. M—marker; 1—non-induced BgGPI52-WH; 2—induced BgGPI52-WH; 3—induced BgGPI52-WH precipitate; 4—induced BgGPI52-WH supernatant; 5—purified recombinant BgGPI52-WH protein.</p> Full article ">Figure 3
<p>The antigenic properties of BgGPI52-WH were detected by Western blot. (<b>a</b>) Immunogenicity detection of <span class="html-italic">B. gibsoni</span> BgGPI52-WH protein. Lane M—marker; lane 1, 3—the iRBC lysates of dogs detected by mouse anti-BgGPI52-WH serum; lane 2, 4—the RBC lysates of uninfected dogs detected by mouse anti-BgGPI52-WH serum. (<b>b</b>) Antigenicity detection of <span class="html-italic">B. gibsoni</span> BgGPI52-WH protein. Lane M—marker; lane 1—recombinant BgGPI52-WH protein detected by the serum of <span class="html-italic">B. gibsoni</span>-infected dogs; Lane 2—recombinant BgGPI52-WH protein detected by normal serum from uninfected dogs.</p> Full article ">Figure 4
<p>Sensitivity of indirect ELISA to BgGPI52-WH. Serum1–Serum5—five known positive sera; cut-off—cut-off value obtained from the above experiment (0.296).</p> Full article ">Figure 5
<p>Indirect ELISA specific test. Bands 1–7 reflect serum from dogs infected with <span class="html-italic">Toxoplasma gondii</span>, <span class="html-italic">Echinococcus granulosus</span>, <span class="html-italic">Strongyloides stercoralis</span>, <span class="html-italic">Rabies virus</span>, <span class="html-italic">Canine Parvovirus</span>, <span class="html-italic">Babesia canis</span>, and <span class="html-italic">Babesia gibsoni</span> (positive control), respectively. Point 8—negative control.</p> Full article ">Figure 6
<p>The antibody change curve of BgGPI52-WH detected by BgGPI52-WH-ELISA. The three experimental beagles infected with <span class="html-italic">B. gibsoni</span> under laboratory conditions are labelled A, B, and C.</p> Full article ">Figure 7
<p>The detection of BgGPI52-WH-ELISA from a clinical sample. The dashed read line indicates the cut-off value. The critical value is X + 3SD = 0.296. Results criteria: serum samples with S/P values greater than 0.296 could be judged as positive. The positive rate after the ELISA test was 11.41% (17/149).</p> Full article ">
14 pages, 1795 KiB  
Article
The Mantle Transcriptome of Chamelea gallina (Mollusca: Bivalvia) and Shell Biomineralization
by Federica Carducci, Maria Assunta Biscotti, Alessandro Mosca, Samuele Greco, Marco Gerdol, Francesco Memmola, Marco Barucca and Adriana Canapa
Animals 2022, 12(9), 1196; https://doi.org/10.3390/ani12091196 - 6 May 2022
Cited by 2 | Viewed by 2437
Abstract
The striped venus Chamelea gallina is a bivalve mollusc that represents one of the most important fishery resources of the Adriatic Sea. In this work, we investigated for the first time the ability of this species to modulate the expression of genes encoding [...] Read more.
The striped venus Chamelea gallina is a bivalve mollusc that represents one of the most important fishery resources of the Adriatic Sea. In this work, we investigated for the first time the ability of this species to modulate the expression of genes encoding proteins involved in biomineralization process in response to biotic and abiotic factors. We provided the first comprehensive transcriptome from the mantle tissue of clams collected in two sampling sites located along the Italian Adriatic coast and characterized by different environmental features. Moreover, the assessment of environmental parameters, scanning electron microscopy (SEM), and X-ray diffraction (XRD) measurements on valves were conducted to better contextualize RNA sequencing (RNA-Seq) data. Functional annotation of differentially expressed genes (DEGs) and SEM observations highlighted a different shell mineralization behaviour in C. gallina clams collected from two selected sites characterized by diverse environmental parameters. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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<p>Geographical localization of sampling sites considered in the present work.</p> Full article ">Figure 2
<p>Circos plot obtained from functional enrichment analysis performed on differentially expressed genes derived from the comparison S vs. SM.</p> Full article ">Figure 3
<p>Scanning electron microscopy (SEM) observations obtained at 500× of magnification. (<b>A</b>–<b>C</b>) are referred to specimens of the Senigallia (S) sampling site. (<b>D</b>–<b>F</b>) are referred to specimens of the Silvi Marina (SM) sampling site.</p> Full article ">Figure 4
<p>Timeseries of environmental parameters. Daily temperature, salinity, and chlorophyll at the sampling stations of Senigallia (S, red line) and Silvi Marina (SM, black line) were reported. Computed monthly means are superimposed as circles.</p> Full article ">
16 pages, 351 KiB  
Article
Creatinine as a Urinary Marker of the Purine Derivatives Excretion in Urine Spot Samples of Lambs Fed Peach Palm Meal
by Taiala Cristina de Jesus Pereira, Mara Lúcia Albuquerque Pereira, Gleidson Giordano Pinto de Carvalho, Herymá Giovane de Oliveira Silva, Alana Batista dos Santos, Douglas dos Santos Pina and Leandro Borges Sousa
Animals 2022, 12(9), 1195; https://doi.org/10.3390/ani12091195 - 6 May 2022
Cited by 1 | Viewed by 2091
Abstract
The objective was to evaluate the influence of diets on lambs using different levels of peach palm meal as a replacement for maize (0, 10, 40, 60, and 85% of diet dry matter) on the endogenous creatinine clearance (CC), urine concentration ratio of [...] Read more.
The objective was to evaluate the influence of diets on lambs using different levels of peach palm meal as a replacement for maize (0, 10, 40, 60, and 85% of diet dry matter) on the endogenous creatinine clearance (CC), urine concentration ratio of purine derivatives to creatinine (PDC index), and daily creatinine excretion (DCE) as a marker to estimate purine derivatives (PD) excretion from urinary spot samples collected at different time points (4, 8, 12, 16, 20, 24 h after morning feeding) compared to 24-h total urine collection. The measured parameters were voluntary intake, urinary volume, CC, DCE, the concentration of plasma creatinine, and PD and purine derivatives’ excretion (PDE). Five lambs were allocated to metabolic cages and distributed in a 5 × 5 Latin square. Urine collection was taken daily on days 16 to 19 of each experimental period. The inclusion of peach palm meal linearly reduced the intake of dry matter (g kg BW−0.75, p = 0.005), crude protein (g kg BW−0.75, p = 0.010), metabolizable energy (MJ kg BW−0.75, p = 0.010) and CC (p < 0.0001). It also quadratically affected the urinary volume (p = 0.008) and DCE (p = 0.004). There was a linear decrease for PDC index (p = 0.032) and PDE (p < 0.0001) measured in the 24-h total urine with peach palm meal levels. The different times of spot urine sampling did not affect (p > 0.05) the PDC index and PDE. Peach palm meal decreases the CC thereby compromising the use of a mean value of DCE as a PDE marker in spot urine samples. There is greater accuracy when using different values of DCE obtained for each diet as markers for the PDE in spot urine samples. Unconventional foodstuffs of low palatability affecting the voluntary intake of feed change the renal function. Full article
(This article belongs to the Section Animal Physiology)
15 pages, 2722 KiB  
Article
The Multi-Omics Analysis Revealed a Metabolic Regulatory System of Cecum in Rabbit with Diarrhea
by Jie Wang, Kaisen Zhao, Zhe Kang, Meigui Wang, Yang Chen, Huimei Fan, Siqi Xia and Songjia Lai
Animals 2022, 12(9), 1194; https://doi.org/10.3390/ani12091194 - 6 May 2022
Cited by 8 | Viewed by 2801
Abstract
With the comprehensive prohibition of antibiotics in the feed industry in China, the incidence of diarrhea in rabbits increased, such as loss of appetite, vomiting, and excretion of atheromatous feces. In order to explore the pathological and the molecular mechanisms of the diarrhea [...] Read more.
With the comprehensive prohibition of antibiotics in the feed industry in China, the incidence of diarrhea in rabbits increased, such as loss of appetite, vomiting, and excretion of atheromatous feces. In order to explore the pathological and the molecular mechanisms of the diarrhea in the rabbitry fed with antibiotic-free diet, we used microbial metagenomics, transcriptome, and non-targeted metabolomics sequencing. The results showed that the Firmicutes level was significantly decreased (p < 0.001) and the Proteobacteria level was significantly increased (p < 0.05). The functional enrichment of cecum revealed that most differentially expressed genes (DEGs) were expressed in immune, inflammatory, and metabolic processes. The enrichment of the cecal fecal metabolites focused on the bile secretion, antifolate resistance, and tryptophan metabolism pathways, which are mainly associated with inflammation. The results of correlation analysis showed that Fournierella was positively correlated with myricetin, ursolic acid, and furtherly might cause bile secretion and tryptophan metabolism disorder, aggravate intestinal inflammation, change intestinal permeability, and reduce host immunity, leading to diarrhea in rabbits. This study provides a theoretical basis for illustrating the reason for diarrhea and developing new feeds for the health of rabbits. Full article
(This article belongs to the Section Animal Physiology)
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Figure 1
<p>Pathological observation of cecum tissue in rabbits by microscope (HE-staining, 100×). (<b>A</b>) Dia_Ce. (<b>B</b>) Con_Ce.</p> Full article ">Figure 2
<p>Comparison of fecal flora in cecum between Dia_Ce and Con_Ce. (<b>A</b>) ANOSIM analysis: R &gt; 0, significant difference in two groups. (<b>B</b>) The number of unique and common OTUs between the two groups shown in the Venn diagram. (<b>C</b>) Relative abundance histogram of species at phylum level. (<b>D</b>) Relative abundance histogram of species at genus level. (<b>E</b>) LDA score of LEfSe–PICRUSt.</p> Full article ">Figure 3
<p>Differentially expressed genes in cecum of diarrhea-afflicted rabbits. (<b>A</b>) Volcano map of DEGs in cecum tissue of Con_Ce and Dia_Ce. (<b>B</b>) GO terms of Dia_Ce vs. Con_Ce. (<b>C</b>) KEGG pathways of Dia_Ce vs. Con_Ce.</p> Full article ">Figure 4
<p>PCA and PLS-DA of fecal metabolites in cecum. (<b>A</b>,<b>B</b>) PCA score graph between Dia_Ce and Con_Ce; (<b>C</b>,<b>D</b>) permutation test from PLS-DA models. Note: (<b>A</b>,C): cationic mode; (<b>B</b>,<b>D</b>): anion mode.</p> Full article ">Figure 5
<p>Differential metabolites and metabolic pathways in cecal feces of diarrhea-afflicted rabbits. (<b>A</b>,<b>B</b>) Volcano of metabolites with significant differences between Dia_Ce and Con_Ce, where A is a positive ion mode and B is a negative ion mode. (<b>C</b>,<b>D</b>) KEGG enrichment bubble diagram, where C is a positive ion mode and D is a negative ion mode.</p> Full article ">Figure 6
<p>Correlation of DM with cecum bacteria and DEGs. (<b>A</b>,<b>B</b>) Correlation analysis results of metabolites top20 and 16S differential bacteria top10. (<b>C</b>,<b>D</b>) KEGG enrichment analysis of metabolism and transcription. Note: (<b>A</b>,<b>C</b>) are positive ion modes; (<b>B</b>,<b>D</b>) are negative ion modes.</p> Full article ">
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