Special Issue "Immunohistochemical and Physiological Research on Farm Animals"

A special issue of Animals (ISSN 2076-2615). This special issue belongs to the section "Animal Physiology".

Deadline for manuscript submissions: closed (28 February 2021).

Special Issue Editors

Prof. Dr. Paola Scocco
E-Mail Website
Guest Editor
School of Biosciences and Veterinary Medicine, University of Camerino (MC), Camerino, Italy
Interests: histochemistry; morphometry; animal welfare; food-intake; digestive apparatus; reproductive apparatus; mammary gland; ruminants; environmental sustainability
Special Issues and Collections in MDPI journals
Dr. Elena De Felice
E-Mail Website
Guest Editor
School of Biosciences and Veterinary Medicine, University of Camerino, 62032 Camerino MC, Italy
Interests: molecular biology; immunohistochemistry; food intake; regulation of food intake; gut-brain axis; aging
Prof. Alessandro Malfatti
E-Mail Website
Guest Editor
School of Biosciences and Veterinary Medicine, University of Camerino (MC), Italy
Interests: animal welfare; animal behavior; sexual hormones; ruminant’s fertility

Special Issue Information

Dear Colleagues,

This Special Issue is dedicated to the application of immunohistochemical and physiological studies carried out on farm animals (swine, sheep, goat, bovine, buffalo, rabbit, poultry, horse, and donkey).

Farm species play very important economic and sociocultural roles, such as food supply, source of income, asset saving, source of employment, soil fertility, livelihood, transport, agricultural traction, agricultural diversification, and sustainable agricultural production. For this reason, it is essential to study in deeper detail the anatomy and physiology of these species. In the last couple of decades, there has been an exponential increase in publications on immunohistochemistry (IHC) and immunocytochemistry techniques, reflecting the currently position that IHC mostly holds in pathological laboratories. This Special Issue will attempt to provide the relevance of immunohistochemical analysis and of their relationships with functions also in farm animals. However, taking into account that multiple methods (Western blot, FISH, etc.) are necessary to perform more reliable conclusions, the use of these methods is also welcomed. Original manuscripts, review articles, as well as short communications are invited. Particularly welcome will be research with a potential applied purpose, studies whose results could be useful in improving farm management practices and the responsible use of natural resources, enhancing the nutraceutical properties of animal derived products, and promoting the circular economy and, in turn, the sustainability of livestock.
You are cordially invited to contribute on this theme or related research topics in order to improve the anatomical and physiological knowledge of farm animals.

Prof. Paola Scocco
Dr. Elena De Felice
Prof. Alessandro Malfatti
Guest Editors

Manuscript Submission Information

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Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Animals is an international peer-reviewed open access monthly journal published by MDPI.

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Keywords

  • farm animals (swine, sheep, goat, bovine, buffalo, rabbit, poultry, horse, donkey)
  • immunohistochemistry and special staining
  • immunohistochemical analysis
  • physiological parameters
  • endocrinology
  • animal behavioral features
  • livestock farming
  • sustainability

Published Papers (20 papers)

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Research

Article
Changes of Plasma Analytes Reflecting Metabolic Adaptation to the Different Stages of the Lactation Cycle in Healthy Multiparous Holstein Dairy Cows Raised in High-Welfare Conditions
Animals 2021, 11(6), 1714; https://doi.org/10.3390/ani11061714 - 08 Jun 2021
Viewed by 736
Abstract
Here, we tested the changes occurring in several plasma analytes during different stages of the lactation cycle of high welfare raised multiparous Holstein cows, and provided reference intervals (RI) for plasma analytes concentrations. Eleven high-welfare farms (HWF) located in Northern Italy were selected [...] Read more.
Here, we tested the changes occurring in several plasma analytes during different stages of the lactation cycle of high welfare raised multiparous Holstein cows, and provided reference intervals (RI) for plasma analytes concentrations. Eleven high-welfare farms (HWF) located in Northern Italy were selected and their herds used to recruit 361 clinically healthy cows undergoing the dry (from −30 to −10 days from real calving; DFC), the postpartum (from 3 to 7 DFC), the early lactation (from 28 to 45 DFC) and the late lactation phases (from 160 to 305 DFC). Cows affected by subclinical diseases (SCD) were retrospectively excluded, and a subset of 285 cows was selected. Data of plasma analytes underwent ANOVA testing using physiological phases as predictors. The individual effect of each phase was assessed using a pairwise t-test assuming p ≤ 0.05 as a significance limit. A bootstrap approach was used to define the reference interval (RI) for each blood analyte within physiological phases having a pairwise t-test p ≤ 0.05. The concentration of nonesterified fatty acids, albumin, cholesterol, retinol, paraoxonase and tocopherol changed throughout all the physiological phases, whereas the concentration of K, alkaline phosphatase and thiol groups remained stable. Triglycerides, Zn, and ferric ion reducing antioxidant power in the dry phase and BHB, Ca, myeloperoxidase, haptoglobin, reactive oxygen metabolites and advanced oxidation of protein product in postpartum differed compared with other physiological phases. During the dry phase, Packed cell volume, Cl, and urea concentrations were similar to during the postpartum phase. Similarly, Na, γ-glutamyl transferase and β-carotene concentrations were similar to during the early lactation phase; fructosamine and bilirubin concentrations were similar to during the late lactation phase. During the postpartum phase, fructosamine and P concentrations were similar to during the early lactation phase, and the aspartate transaminase concentration was similar to during the late lactation phase. During the early lactation phase, Mg, creatinine, total protein, globulin and ceruloplasmin concentrations were similar to during the postpartum phase, while the urea concentration was similar to during the late lactation phase. All these plasma analytes differed among the other phases. This study identifies physiological trends affecting plasma analytes concentrations during the different stages of the lactation cycle and provides a guideline for the duration and magnitude of their changes when animals are healthy and raised in optimal welfare conditions. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
A Prototype Skin Substitute, Made of Recycled Marine Collagen, Improves the Skin Regeneration of Sheep
Animals 2021, 11(5), 1219; https://doi.org/10.3390/ani11051219 - 23 Apr 2021
Viewed by 589
Abstract
Skin wound healing is a complex and dynamic process that aims to restore lesioned tissues. Collagen-based skin substitutes are a promising treatment to promote wound healing by mimicking the native skin structure. Recently, collagen from marine organisms has gained interest as a source [...] Read more.
Skin wound healing is a complex and dynamic process that aims to restore lesioned tissues. Collagen-based skin substitutes are a promising treatment to promote wound healing by mimicking the native skin structure. Recently, collagen from marine organisms has gained interest as a source for producing biomaterials for skin regenerative strategies. This preliminary study aimed to describe the application of a collagen-based skin-like scaffold (CBSS), manufactured with collagen extracted from sea urchin food waste, to treat experimental skin wounds in a large animal. The wound-healing process was assessed over different time points by the means of clinical, histopathological, and molecular analysis. The CBSS treatment improved wound re-epithelialization along with cell proliferation, gene expression of growth factors (VEGF-A), and development of skin adnexa throughout the healing process. Furthermore, it regulated the gene expression of collagen type I and III, thus enhancing the maturation of the granulation tissue into a mature dermis without any signs of scarring as observed in untreated wounds. The observed results (reduced inflammation, better re-epithelialization, proper development of mature dermis and skin adnexa) suggest that sea urchin-derived CBSS is a promising biomaterial for skin wound healing in a “blue biotechnologies” perspective for animals of Veterinary interest. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Effects of Feed Supplementation on Nesfatin-1, Insulin, Glucagon, Leptin, T3, Cortisol, and BCS in Milking Ewes Grazing on Semi-Natural Pastures
Animals 2021, 11(3), 682; https://doi.org/10.3390/ani11030682 - 04 Mar 2021
Viewed by 402
Abstract
This study aimed to investigate the effects of feed supplementation on body condition score (BCS) and different metabolic hormones concentration in lactating sheep reared in Italian Central Apennine pastures during the grazing summer period. In this study, 24 Comisana x Appenninica pluriparous ewes [...] Read more.
This study aimed to investigate the effects of feed supplementation on body condition score (BCS) and different metabolic hormones concentration in lactating sheep reared in Italian Central Apennine pastures during the grazing summer period. In this study, 24 Comisana x Appenninica pluriparous ewes from June until August were divided into two homogenous groups: the control group (UNS) was free to graze, while the other group (SUP), in addition to grazing, was supplemented with 600 g/day/head of cereals. At the start of the supplementation and at an interval of 9–10 days until the end of experimentation, BCS evaluation and blood withdrawal to assay nesfatin-1, insulin, glucagon, leptin, triiodothyronine and cortisol levels were performed. Univariable analysis showed no remarkable differences between the groups, while multivariable analysis suggested that the UNS group was characterized by a lower BCS and greater nesfatin-1 than the SUP group. These findings can be considered in relation to milk production, which shows a clear better persistence in the SUP group. Our results indicate that nutritional supplementation has protected ewes from the usual lowering of the body state linked to lactation and provides a good maintenance of milk production, determining also a better overall body and metabolic state of the animals, which is important at the beginning of the sexual season. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Approaches to Identify Pregnancy Failure in Buffalo Cows
Animals 2021, 11(2), 487; https://doi.org/10.3390/ani11020487 - 12 Feb 2021
Viewed by 833
Abstract
The aim of this work was to find the best strategy to diagnose pregnancy failures in buffalo. A total of 109 animals belonging to a buffalo herd subjected to a synchronization and artificial insemination (AI) program were enrolled in this study. Blood samples [...] Read more.
The aim of this work was to find the best strategy to diagnose pregnancy failures in buffalo. A total of 109 animals belonging to a buffalo herd subjected to a synchronization and artificial insemination (AI) program were enrolled in this study. Blood samples were collected at days 0, 14, 25, 28 and 40 after AI for the determination of progesterone (P4) and pregnancy-associated glycoproteins (PAGs) by the radioimmunoassay (RIA) method. Transrectal ultrasonography was performed on day 25, 28 and 40 after AI to monitor pregnancy. The animals included in the data analysis were assigned ex post in pregnant (n = 50) and mortality (n = 12) groups. By ultrasonography, the predictive sign of mortality was the heartbeat. At day 25, the PAGs concentration was significant in predicting embryonic mortality with respect to ultrasonography and P4, at the cut-off of 1.1 ng/mL. At day 28, either PAGs, at a cut-off of 2.2 ng/mL, or ultrasonography, with no detection of heartbeat, were highly predictive of embryonic mortality. PAGs were the best marker (p < 0.05) for predicting embryonic mortality between 25 and 40 days of gestation in buffalo. Its utilization as a diagnostic tool can influence management decisions in order to improve farm reproductive management. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Distinct Pattern of NPY in Gastro–Entero–Pancreatic System of Goat Kids Fed with a New Standardized Red Orange and Lemon Extract (RLE)
Animals 2021, 11(2), 449; https://doi.org/10.3390/ani11020449 - 09 Feb 2021
Cited by 2 | Viewed by 596
Abstract
The use of natural compounds as feed additive is also increasing in farm animals, thanks to the beneficial effect on both animals and consumers health. Here, we questioned whether natural extracts, such as red orange and lemon extract (RLE) rich in flavanones, anthocyanins, [...] Read more.
The use of natural compounds as feed additive is also increasing in farm animals, thanks to the beneficial effect on both animals and consumers health. Here, we questioned whether natural extracts, such as red orange and lemon extract (RLE) rich in flavanones, anthocyanins, and other polyphenols, used as feed additives could display an effect on the neuropeptide Y (NPY) in the gastro–entero–pancreatic tract of goat kids. NPY is one of the most abundant neuropeptides in mammals, known for its orexigenic role although it is involved in many central and peripheral functions. We carried out immunohistochemical analyses on samples of abomasum, duodenum and pancreas collected from two experimental groups: one fed with standard diet and one with standard diet + RLE. For the first time we document NPY distribution in the abomasum, duodenum and pancreas of goats and observe the highest number of NPY positive cells in neuroendocrine cells of duodenum. Remarkably, upon RLE feed supplementation, NPY immunoreactive cells increased significantly in abomasal epithelium and pancreatic islets but not in duodenum, likely due to pH variation of abomasum and duodenum. Our observations represent a baseline for future studies on the interaction between neuropeptides and polyphenols, used as feed additive. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Cannabinoid and Cannabinoid-Related Receptors in the Myenteric Plexus of the Porcine Ileum
Animals 2021, 11(2), 263; https://doi.org/10.3390/ani11020263 - 21 Jan 2021
Viewed by 668
Abstract
An important piece of evidence has shown that molecules acting on cannabinoid receptors influence gastrointestinal motility and induce beneficial effects on gastrointestinal inflammation and visceral pain. The aim of this investigation was to immunohistochemically localize the distribution of canonical cannabinoid receptor type 1 [...] Read more.
An important piece of evidence has shown that molecules acting on cannabinoid receptors influence gastrointestinal motility and induce beneficial effects on gastrointestinal inflammation and visceral pain. The aim of this investigation was to immunohistochemically localize the distribution of canonical cannabinoid receptor type 1 (CB1R) and type 2 (CB2R) and the cannabinoid-related receptors transient potential vanilloid receptor 1 (TRPV1), transient potential ankyrin receptor 1 (TRPA1), and serotonin receptor 5-HT1a (5-HT1aR) in the myenteric plexus (MP) of pig ileum. CB1R, TRPV1, TRPA1, and 5-HT1aR were expressed, with different intensities in the cytoplasm of MP neurons. For each receptor, the proportions of the immunoreactive neurons were evaluated using the anti-HuC/HuD antibody. These receptors were also localized on nerve fibers (CB1R, TRPA1), smooth muscle cells of tunica muscularis (CB1R, 5-HT1aR), and endothelial cells of blood vessels (TRPV1, TRPA1, 5-HT1aR). The nerve varicosities were also found to be immunoreactive for both TRPV1 and 5-HT1aR. No immunoreactivity was documented for CB2R. Cannabinoid and cannabinoid-related receptors herein investigated showed a wide distribution in the enteric neurons and nerve fibers of the pig MP. These results could provide an anatomical basis for additional research, supporting the therapeutic use of cannabinoid receptor agonists in relieving motility disorders in porcine enteropathies. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Communication
Epithelial to Mesenchymal Transition (EMT) in a Laryngeal Squamous Cell Carcinoma of a Horse: Future Perspectives
Animals 2020, 10(12), 2318; https://doi.org/10.3390/ani10122318 - 07 Dec 2020
Viewed by 754
Abstract
Squamous cell carcinoma (SCC) is one of the most frequent tumors of skin and muco-cutaneous junctions in the horse. Equine papillomavirus type 2 (EcPV2) has been detected in equine SCC of the oral tract and genitals, and recently also in the larynx. As [...] Read more.
Squamous cell carcinoma (SCC) is one of the most frequent tumors of skin and muco-cutaneous junctions in the horse. Equine papillomavirus type 2 (EcPV2) has been detected in equine SCC of the oral tract and genitals, and recently also in the larynx. As human squamous cell carcinoma of the larynx (SCCL), it is strongly etiologically associated with high-risk papillomavirus (h-HPV) infection. This study focuses on tumor cells behavior in a naturally occurring tumor that can undergo the so-called epithelial to mesenchymal transition (EMT). A SCCL in a horse was investigated by immunohistochemistry using antibodies against E-cadherin, pan-cytokeratin AE3/AE1, β-catenin, N-cadherin, vimentin, ZEB-1, TWIST, and HIF-1α. EcPV2 DNA detection and expression of oncogenes in SCC were investigated. A cadherin switch and an intermediate filaments rearrangement within primary site tumor cells together with the expression of the EMT-related transcription factors TWIST-1, ZEB-1, and HIF-1α were observed. DNA obtained from the tumor showed EcPV2 positivity, with E2 gene disruption and E6 gene dysregulation. The results suggest that equine SCCL might be a valuable model for studying EMT and the potential interactions between EcPV2 oncoproteins and the EMT process in SCCL. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Effects of Aging on Expression of Mic60 and OPA1 and Mitochondrial Morphology in Myocardium of Tibetan Sheep
Animals 2020, 10(11), 2160; https://doi.org/10.3390/ani10112160 - 20 Nov 2020
Viewed by 565
Abstract
In order to investigate the effects of aging on the expression of Mic60 and OPA1 and mitochondrial morphology in plateau animals, the expression of Mic60 and OPA1 genes and proteins, and the morphology of mitochondria in the myocardium of adult and aged Tibetan [...] Read more.
In order to investigate the effects of aging on the expression of Mic60 and OPA1 and mitochondrial morphology in plateau animals, the expression of Mic60 and OPA1 genes and proteins, and the morphology of mitochondria in the myocardium of adult and aged Tibetan sheep were investigated. The expression of Mic60 and OPA1 genes and OPA1 protein were higher (p < 0.05) in the myocardium of adult Tibetan sheep than in those of the aged ones. The number of mitochondrial cristae in the myocardium of adult was higher than that in aged (p < 0.05). The density of mitochondria in the myocardium of adult was higher than that in aged (p < 0.01). Compared with the adult Tibetan sheep, the mitochondrial crista of aged were relatively sparse, the crista membrane was wide, and the mitochondria were not closely linked, showing fragmentation. These results suggest that the myocardial mitochondria of the adult have better energy supply ability, indicating that aging can lead to the weakening of oxygen supply in the myocardial mitochondria of Tibetan sheep. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Productive and Physiological Response of Male Rabbits to Dietary Supplementation with Thyme Essential Oil
Animals 2020, 10(10), 1844; https://doi.org/10.3390/ani10101844 - 10 Oct 2020
Cited by 4 | Viewed by 622
Abstract
The present study aimed at assessing the efficiency of thyme essential oil (TEO) as an alternative to antibiotics for improving the productive performance and serum metabolic profile of male rabbits. A total of one hundred and fifty 70-day-old male Californian rabbits were assigned [...] Read more.
The present study aimed at assessing the efficiency of thyme essential oil (TEO) as an alternative to antibiotics for improving the productive performance and serum metabolic profile of male rabbits. A total of one hundred and fifty 70-day-old male Californian rabbits were assigned to five dietary treatments, including a basal diet as a negative control (NC), a basal diet supplemented with an antibiotic as a positive control (PC), and a basal diet supplemented with 60, 120, or 180 mg/kg of TEO. The experiment period lasted for 60 days. Supplementation of TEO levels significantly (P < 0.01) increased daily body weight gain and improved feed conversion ratio of male Californian rabbits compared to NC and PC groups. Similarly, the TEO remarkably enhanced the semen characteristics of rabbits compared to NC and PC groups. Supplementation of TEO significantly decreased aspartate transaminase, alanine transaminase, urea, and creatinine compared with NC and PC groups. Supplementation with TEO increased serum testosterone concentration compared to NC and PC treatments. Our data demonstrate that TEO levels up to 180 mg/kg can play a major role as an alternative to dietary antibiotics, in improving the productive performance, semen quality, testosterone levels, and the kidney and liver functions in California male rabbits. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Plasmatic Profile of Pregnancy-Associated Glycoprotein (PAG) during Gestation and Postpartum in Sarda and Lacaune Sheep Determined with Two Radioimmunoassay Systems
Animals 2020, 10(9), 1502; https://doi.org/10.3390/ani10091502 - 25 Aug 2020
Cited by 1 | Viewed by 560
Abstract
This study was carried out to determine ovine pregnancy-associated glycoprotein (oPAG) levels in the plasma of Sarda and Lacaune ewes throughout gestation and in the first month postpartum, using two heterologous radioimmunoassays (RIA-706 and RIA-srPool) and to study the correlations between PAG levels [...] Read more.
This study was carried out to determine ovine pregnancy-associated glycoprotein (oPAG) levels in the plasma of Sarda and Lacaune ewes throughout gestation and in the first month postpartum, using two heterologous radioimmunoassays (RIA-706 and RIA-srPool) and to study the correlations between PAG levels and fetal gender and number. On Day 18 of pregnancy, PAG concentrations were detected in 90.1% and 80.8% of Sarda pregnant ewes with RIA-706 and RIA-srPool, respectively; and in 90% and in 75% of Lacaune pregnant ewes with RIA-706 and RIA-srPool, respectively. From Day 30, PAG concentrations were detected in all pregnant ewes by using both RIA methods. In the postpartum period, the PAG concentrations in Sarda ewes decreased rapidly reaching minimal levels (<1 ng/mL) on day 28 using both RIA-706 and RIA-srPool. In Lacaune ewes, PAG-706 levels were higher than PAG-srPool from parturition until the last day of observation (Day 28 postpartum). It was also observed that mean concentrations were higher in multiple than in single pregnancies in Sarda and Lacaune ewes. Moreover, due to the rapid disappearance of PAG concentration following lambing, a cut-off limit in postpartum animals was not required as a means for detecting a new pregnancy. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Communication
Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Exhibits High Predicted Binding Affinity to ACE2 from Lagomorphs (Rabbits and Pikas)
Animals 2020, 10(9), 1460; https://doi.org/10.3390/ani10091460 - 20 Aug 2020
Cited by 4 | Viewed by 1118
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the pandemic COVID-19. The virus infects human cells by binding of the virus spike to the cell receptor ACE2. The crystal structure of SARS-CoV-2 spikes in complex with human ACE2 has recently been [...] Read more.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is responsible for the pandemic COVID-19. The virus infects human cells by binding of the virus spike to the cell receptor ACE2. The crystal structure of SARS-CoV-2 spikes in complex with human ACE2 has recently been solved, and the main amino acid residues involved in the virus–receptor complex have been detected. To investigate the affinity of ACE2 of lagomorphs to the SARS-CoV-2 spike, ACE2 sequences from rabbits and American pikas were compared with human ACE2 and with ACE2 from mammals with different susceptibility to the virus. Models of the complex formed by SARS-CoV-2 spike and ACE2 from lagomorphs and from other mammals were created for comparative studies. ACE2 of lagomorphs showed fewer substitutions than human ACE2 in residues involved in the ACE2-SARS-CoV-2 spike complex, similar to cats. Analysis of the binding interface of the simulated complexes ACE2-SARS-CoV-2 spike showed high affinity of the ACE2 of lagomorphs to the viral spike protein. These findings suggest that the spike of SARS-CoV-2 could bind the ACE2 receptor of lagomorphs, and future studies should investigate the role of lagomorphs in SARS-CoV-2 epidemiology. Furthermore, the risks to humans coming into close contacts with these animals should be evaluated. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Productive Results, Oxidative Stress and Contaminant Markers in European Sea Bass: Conventional vs. Organic Feeding
Animals 2020, 10(7), 1226; https://doi.org/10.3390/ani10071226 - 18 Jul 2020
Viewed by 681
Abstract
In the present study European sea bass (Dicentrarchus labrax) subjected to two different diets (organic vs. conventional) were evaluated in terms of growing performances, oxidative stress, and contaminant markers. Growing performances were evaluated using biometric measures and condition factor (K), whereas [...] Read more.
In the present study European sea bass (Dicentrarchus labrax) subjected to two different diets (organic vs. conventional) were evaluated in terms of growing performances, oxidative stress, and contaminant markers. Growing performances were evaluated using biometric measures and condition factor (K), whereas insulin-like growth factor (IGF-I and IGF-II) levels were assessed trough Real-Time PCR analysis. For oxidative stress, immunohistochemical staining for 8-hydroxy-2′-deoxyguanosine (8-OHdG) and 4-hydroxy-2-nonenal (HNE) was performed, whereas total glutathione (GSH) in blood serum was determined by an enzymatic method adapted. Cytochrome P4501A (CYP1A) and melanomacrophage centers (MMCs) were evaluated as contaminant markers trough immunohistochemical and histochemical approaches, respectively. The growing performances showed a positive trend in both groups but a greater productivity in conventional fed fish compared to the organic ones. A significant higher expression of MMCs was observed in organic vs. conventional diet fed fish. Fillet analysis showed a higher MUFA content and a lower PUFAs n-6 content in organically fed sea bass indicating that diets with a content in fatty acids closer to that of wild fish will definitely affect the fatty acid profile of the fish flesh. On the other hand, the diet composition did not seem to affect neither the oxidative stress parameters (GSH, 8-OHdG, HNE) nor the CYP1A expression. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Immunohistochemical Analysis of Intestinal and Central Nervous System Morphology in an Obese Animal Model (Danio rerio) Treated with 3,5-T2: A Possible Farm Management Practice?
Animals 2020, 10(7), 1131; https://doi.org/10.3390/ani10071131 - 03 Jul 2020
Cited by 1 | Viewed by 675
Abstract
The 3,5-diiodo-L-thyronine (3,5-T2) is an endogenous metabolite of thyroid hormones, whose administration to rodents fed high-fat diet (HFD) prevents body weight increase and reverts the expression pattern of pro-inflammatory factors associated to HFD. The diet-induced obese (D.I.O.) zebrafish (Danio rerio) has been recently [...] Read more.
The 3,5-diiodo-L-thyronine (3,5-T2) is an endogenous metabolite of thyroid hormones, whose administration to rodents fed high-fat diet (HFD) prevents body weight increase and reverts the expression pattern of pro-inflammatory factors associated to HFD. The diet-induced obese (D.I.O.) zebrafish (Danio rerio) has been recently used as an experimental model to investigate fundamental processes underlying central and peripheral obesity-driven inflammation. Herein, we aim to understand the role of 3,5-T2 in regulating central and peripheral inflammation in D.I.O. model of zebrafish. 3,5-T2 (10 nM and 100 nM) was administered with the obesity-inducing diet (D.I.O. with 3,5-T2) or after 4 weeks of obesity-inducing diet (D.I.O. flw 3,5-T2). 3,5-T2 significantly increased the body weight and serum triglyceride levels in D.I.O. zebrafish in both conditions. Moreover, 3,5-T2 sustained or increased inflammation in the anterior (AI) and mid (MI) intestine when administered with the obesity-inducing diet, as indicated by the immunoexpression of the inflammatory markers tumor-necrosis factor-α (TNFα), cyclooxygenase 2 (COX2), calnexin, caspase 3, and proliferating cell nuclear antigen (PCNA). On the contrary, when 3,5-T2 was administered after the obesity-inducing diet, partly reverted the intestinal alteration induced by D.I.O. In addition, brain inflammation, as indicated by the increase in the activation of microglia, was detected in D.I.O. zebrafish and D.I.O. treated with 3,5-T2. These findings reveal that the effects of 3,5-T2 on fish intestine and brain can deviate from those shown in obese mammals, opening new avenues to the investigation of the potential impact of this thyroid metabolite in different diseases including obesity. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Influence of Different Feed Physical Forms on Mandibular Gland in Growing Pigs
Animals 2020, 10(5), 910; https://doi.org/10.3390/ani10050910 - 24 May 2020
Cited by 1 | Viewed by 855
Abstract
A study was performed on the mandibular gland obtained from growing pigs enrolled in a wide research project aiming to test the effects of different feed physical forms on animal health, production and welfare. We used 48 pigs fed for four weeks with [...] Read more.
A study was performed on the mandibular gland obtained from growing pigs enrolled in a wide research project aiming to test the effects of different feed physical forms on animal health, production and welfare. We used 48 pigs fed for four weeks with different dietary treatments based on different grinding intensities and compactions of the same diet, namely coarsely ground meal (CM), finely ground pelleted (FP) and coarsely ground pelleted (CP) diets. Samples were analyzed by conventional histochemistry to identify the glycohistochemical profile and by immunohistochemistry to localize aquaporin 5, apelin and apelin receptor. Statistical elaborations were performed using the Stats R-package, version 3.5.3. Pig mandibular gland adenomere increased both the quantity and acidity of produced glycoconjugates from CM to FP and CP diets. This probably calls forth higher watery saliva, thus promoting a better feed softening facilitating the amalgamation of the bolus. Mandibular gland increased aquaporin 5 positivity in the CP diet, supporting the hypothesis of an augmented demand for water. Based on apelin/receptor localization, it was hypothesized that in pig mandibular gland the apelinergic system likely performs an endocrine control on the demilunes activity and a paracrine control on ducts, facilitating the production of a more fluid saliva. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Endometritis Changes the Neurochemical Characteristics of the Caudal Mesenteric Ganglion Neurons Supplying the Gilt Uterus
Animals 2020, 10(5), 891; https://doi.org/10.3390/ani10050891 - 20 May 2020
Cited by 3 | Viewed by 716
Abstract
This study analyzed the influence of uterine inflammation on the neurochemical characteristics of the gilt caudal mesenteric ganglion (CaMG) uterus-supplying neurons. The horns of uteri were injected with retrograde tracer Fast Blue on day 17 of the first studied estrous cycle. Twenty-eight days [...] Read more.
This study analyzed the influence of uterine inflammation on the neurochemical characteristics of the gilt caudal mesenteric ganglion (CaMG) uterus-supplying neurons. The horns of uteri were injected with retrograde tracer Fast Blue on day 17 of the first studied estrous cycle. Twenty-eight days later (the expected day 3 of the third studied estrous cycle), either saline or Escherichia coli suspension were administered into each uterine horn. Only the laparotomy was done in the control gilts. After 8 days, the CaMGs and uteri were harvested. The infected gilts presented a severe acute endometritis. In the CaMGs, the populations of uterine perikarya possessing dopamine-β-hydroxylase (DβH) and/or neuropeptide Y (NPY), somatostatin (SOM), galanin (GAL) and vasoactive intestinal polypeptide (VIP) were analyzed using the double immunofluorescence method. In the CaMG, bacterial injection decreased the total number of the perikarya (Fast Blue-positive), the small and large perikarya populations in the dorsal and central regions, and the small and large perikarya populations coded DβH+/GAL- and DβH-/NPY+. After bacterial treatment, there was an increase in the numbers of small and large perikarya coded DβH+/NPY+, small perikarya coded DβH+/GAL+ and DβH+/SOM- and large perikarya coded DβH+/VIP+. To summarize, uterine inflammation influences the neurochemical characteristics of the CaMG uterus-supplying neurons, which may be important for pathologically changed organ functions. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
A Detailed Study of Rainbow Trout (Onchorhynchus mykiss) Intestine Revealed That Digestive and Absorptive Functions Are Not Linearly Distributed along Its Length
Animals 2020, 10(4), 745; https://doi.org/10.3390/ani10040745 - 24 Apr 2020
Cited by 9 | Viewed by 1236
Abstract
To increase the sustainability of trout farming, the industry requires alternatives to fish-based meals that do not compromise animal health and growth performances. To develop new feeds, detailed knowledge of intestinal morphology and physiology is required. We performed histological, histochemical, immunohistochemical and morphometric [...] Read more.
To increase the sustainability of trout farming, the industry requires alternatives to fish-based meals that do not compromise animal health and growth performances. To develop new feeds, detailed knowledge of intestinal morphology and physiology is required. We performed histological, histochemical, immunohistochemical and morphometric analysis at typical time points of in vivo feeding trials (50, 150 and 500 g). Only minor changes occurred during growth whereas differences characterized two compartments, not linearly distributed along the intestine. The first included the pyloric caeca, the basal part of the complex folds and the villi of the distal intestine. This was characterized by a significantly smaller number of goblet cells with smaller mucus vacuoles, higher proliferation and higher apoptotic rate but a smaller extension of fully differentiated epithelial cells and by the presence of numerous pinocytotic vacuolization. The second compartment was formed by the proximal intestine and the apical part of the posterior intestine complex folds. Here we observed more abundant goblet cells with bigger vacuoles, low proliferation rate, few round apoptotic cells, a more extended area of fully differentiated cells and no pinocytotic vacuoles. Our results suggest that rainbow trout intestine is physiologically arranged to mingle digestive and absorptive functions along its length. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Diurnal Rhythm of Plasma Melatonin Concentration in the Domestic Turkey and Its Regulation by Light and Endogenous Oscillators
Animals 2020, 10(4), 678; https://doi.org/10.3390/ani10040678 - 13 Apr 2020
Cited by 1 | Viewed by 639
Abstract
The aim of this study was to characterize the diurnal rhythm of plasma melatonin (MLT) concentration and its regulation by light and endogenous oscillators in 10-week-old domestic turkeys. Three experiments were performed to examine (i) the course of daily changes in plasma MLT [...] Read more.
The aim of this study was to characterize the diurnal rhythm of plasma melatonin (MLT) concentration and its regulation by light and endogenous oscillators in 10-week-old domestic turkeys. Three experiments were performed to examine (i) the course of daily changes in plasma MLT concentration in turkeys kept under a 12 h light: 12 h dark (12L:12D) cycle; (ii) the influence of night-time light exposure lasting 0.5, 1, 2, or 3 h on the plasma MLT level; and (iii) the occurrence of circadian fluctuations in plasma MLT levels in birds kept under continuous dim red light and the ability of turkeys to adapt their pineal secretory activity to a reversed light-dark cycle (12D:12L). The plasma MLT concentration was measured with a direct radioimmunoassay. The plasma MLT concentration in turkeys kept under a 12L:12D cycle changed significantly in a daily rhythm. It was low during the photophase and increased stepwise after the onset of darkness to achieve the maximal level in the middle of the scotophase. Next, it decreased during the second half of the night. The difference between the lowest level of MLT and the highest level was approximately 18-fold. The exposure of turkeys to light during the scotophase caused a rapid, large decrease in plasma MLT concentration. The plasma MLT concentration decreased approximately 3- and 10-fold after 0.5 and 1 h of light exposure, respectively, and reached the day-time level after 2 h of exposure. In turkeys kept under continuous darkness, the plasma MLT level was approximately 2.5-fold higher at 02:00 h than at 14:00 h. In birds kept under 12D:12L, the plasma MLT level was significantly higher at 14:00 h than at 02:00 h. The results showed that plasma MLT concentrations in 10-week-old turkeys have a prominent diurnal rhythm, which is endogenously generated and strongly influenced by environmental light. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Effect of Acrylamide Supplementation on the CART-, VAChT-, and nNOS-Immunoreactive Nervous Structures in the Porcine Stomach
Animals 2020, 10(4), 555; https://doi.org/10.3390/ani10040555 - 26 Mar 2020
Cited by 2 | Viewed by 675
Abstract
Acrylamide is found in food products manufactured with high-temperature processing, and exposure to acrylamide contained in food products may cause a potential risk to human health. The aim of this investigation was to demonstrate the changes in the population of CART-, nNOS-, and [...] Read more.
Acrylamide is found in food products manufactured with high-temperature processing, and exposure to acrylamide contained in food products may cause a potential risk to human health. The aim of this investigation was to demonstrate the changes in the population of CART-, nNOS-, and VAChT-immunoreactive enteric neurons in the porcine stomach in response to supplementation of low and high acrylamide doses. The study was carried out with 15 Danish landrace gilts divided into three experimental groups: the control group—animals were administered empty gelatine capsules; the low-dose group—animals were administrated a tolerable daily intake (TDI) dose (0.5 µg/kg of body weight (b.w.)/day) of acrylamide capsules, and the high-dose group—animals were administrated high-dose (ten times higher than TDI: 5 µg/kg b.w./day) acrylamide capsules for 28 days. Using the double immunofluorescence staining method, it was established that supplementation with low and high doses of acrylamide resulted in alterations of the porcine stomach neuron phenotype, which was reflected in an increased number of CART-, VAChT-, and nNOS-immunoreactive neurons. These changes were accompanied by an increased density of CART-, VAChT-, and nNOS-positive fibres. The results suggest that the enteric nervous system plays an important role in protecting the gastrointestinal tract during acrylamide intoxication. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
Impaired Mitochondrial Function Results from Oxidative Stress in the Full-Term Placenta of Sows with Excessive Back-Fat
Animals 2020, 10(2), 360; https://doi.org/10.3390/ani10020360 - 23 Feb 2020
Cited by 1 | Viewed by 824
Abstract
The aim of this study was to determine the effect of excessive back-fat (BF) of sows on placental oxidative stress, ATP generation, mitochondrial alterations in content and structure, and mitochondrial function in isolated trophoblasts. Placental tissue was collected by vaginal delivery from BFI [...] Read more.
The aim of this study was to determine the effect of excessive back-fat (BF) of sows on placental oxidative stress, ATP generation, mitochondrial alterations in content and structure, and mitochondrial function in isolated trophoblasts. Placental tissue was collected by vaginal delivery from BFI (15–20 mm, n = 10) and BFII (21–27 mm, n = 10) sows formed according to BF at mating. Our results demonstrated that excessive back-fat contributed to augmented oxidative stress in term placenta, as evidenced by excessive production of ROS, elevated protein carbonylation, and reduced SOD, GSH-PX, and CAT activities (p < 0.05). Indicative of mitochondrial dysfunction, reduced mitochondrial respiration in cultured trophoblasts was linked to decreased ATP generation, lower mitochondrial Complex I activity and reduced expression of electron transport chain subunits in placenta of BFII sows (p < 0.05). Meanwhile, we observed negative alterations in mitochondrial biogenesis and structure in the placenta from BFII group (p < 0.05). Finally, our in vitro studies showed lipid-induced ROS production resulted in mitochondrial alterations in trophoblasts, and these effects were blocked by antioxidant treatment. Together, these data reveal that excessive back-fat aggravates mitochondrial injury induced by increased oxidative stress in pig term placenta, which may have detrimental consequences on placental function and therefore impaired fetal growth and development. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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Article
The Expression of ERK1/2 in Female Yak (Bos grunniens) Reproductive Organs
Animals 2020, 10(2), 334; https://doi.org/10.3390/ani10020334 - 20 Feb 2020
Viewed by 751
Abstract
The main reproductive organs undergo different histological appearances and physiological processes under different reproductive statuses. The variation of these organs depends on a delicate regulation of cell proliferation, differentiation, and apoptosis. Extracellular signal-regulated kinases1/2 (ERK1/2) are members of the mitogen-activated protein kinase (MAPK) [...] Read more.
The main reproductive organs undergo different histological appearances and physiological processes under different reproductive statuses. The variation of these organs depends on a delicate regulation of cell proliferation, differentiation, and apoptosis. Extracellular signal-regulated kinases1/2 (ERK1/2) are members of the mitogen-activated protein kinase (MAPK) super family. They have important roles in regulating various biological processes of different cells, tissues, and organ types. Activated ERK1/2 generally promotes cell survival, but under certain conditions, ERK1/2 also have the function of inducing apoptosis. It is widely believed that ERK1/2 play a significant role in regulating the reproductive processes of mammals. The goal of our research is to investigate the expression and distribution of ERK1/2 in the yak’s main reproductive organs during different stages. In the present study, samples of the ovary, oviduct, and uterus of 15 adult female yak were collected and used in the experiment. The ERK1/2 proteins, localization, and quantitative expression of their mRNA were investigated using immunohistochemistry (IHC), western blot (WB) and relative quantitative real-time polymerase chain reaction (RT-PCR). The results indicated that ERK1/2 proteins and their mRNA were highly expressed in the ovary of the luteal phase and gestation period, in the oviduct of the luteal phase, and in the uterus of the luteal phase and gestation period. Immunohistochemical analysis revealed a strong distribution of ERK1/2 proteins in follicular granulosa cells, granular luteal cells, villous epithelial cells of the oviduct, endometrial glandular epithelium, and luminal epithelium. These results demonstrated that the expression of ERK1 and ERK2 proteins and their mRNA in the yak’s ovary, oviduct, and uterus varies with the stage of the reproductive cycle. The variation character of ERK1 and ERK 2 expression in the yak’s main reproductive organs during different stages implies that they play an important role in regulating the reproductive function under different physiological statuses. Full article
(This article belongs to the Special Issue Immunohistochemical and Physiological Research on Farm Animals)
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