Animals

17 pages, 4032 KiB

Open AccessArticle

Gradual Thickening of the Peritubular Lamina Propria in Healthy Boar Seminiferous Tubules Due to Cryptorchidism: Increased Immunoexpression of Diverse Proteins in Sertoli and Myoid Cells

by Vicente Seco-Rovira, Jesús Martínez-Hernández, Laís Freire-Brito, Ester Beltrán-Frutos, Juan Francisco Madrid and Luis Miguel Pastor

Animals 2025, 15(12), 1696; https://doi.org/10.3390/ani15121696 (registering DOI) - 8 Jun 2025

Abstract

In testicular pathology, the accumulation of extracellular matrix and thickening of the peritubular lamina propria are associated with altered spermatogenesis. This study evaluated the immunoexpression of certain proteins in Sertoli and myoid cells of cryptorchid boar seminiferous tubule sections to determine the role [...] Read more.

In testicular pathology, the accumulation of extracellular matrix and thickening of the peritubular lamina propria are associated with altered spermatogenesis. This study evaluated the immunoexpression of certain proteins in Sertoli and myoid cells of cryptorchid boar seminiferous tubule sections to determine the role of these cells in peritubular lamina propria thickening. Sections of normal seminiferous tubules and those with varying degrees of atrophy (I–III) were studied using histochemical and immunohistochemical techniques. A semiquantitative analysis of histochemical staining or immunostaining intensity in these sections was performed. Gradual thickening of the peritubular lamina propria was observed between stage II and III sections. HSP47 was present in myoid and Sertoli cells in normal sections and was significantly increased in stage II and III sections. The increase in stage II was dependent on Sertoli cells, while in stage III, it depended on myoid cells. The vimentin increases in stage I sections were due to Sertoli cells and later to myoid cells in stages II and III. α-actin and collagen IV immunoreactivity were observed from the early stage of atrophy, with a significant increase in stage III. In conclusion, in porcine spontaneous cryptorchidism, the seminiferous tubule exhibits gradual fibrotic alterations in the peritubular lamina propria, initially related to changes in the Sertoli cell phenotype and later, in the final stages, to myoid cells. Collagen I deposition appears to be caused by myoid cells, initiating the sclerosis of the seminiferous tubules. Full article

(This article belongs to the Section Animal Reproduction)

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19 pages, 1319 KiB

Open AccessArticle

Microsporidia in Rodents—Mus musculus, Rattus norvegicus, and Rattus rattus—A Public Health Concern in the Canary Islands, Spain

by Sergio Llorens-Berzosa, Edgar Baz-González, Natalia Martin-Carrillo, Katherine García-Livia, Virginia Amaro-Ramos, Néstor Abreu-Acosta, Carmen del Aguila, Jordi Miquel, Román Pino-Vera, Estefanía Abreu-Yanes, Carlos Feliu, Fernando Izquierdo and Pilar Foronda

Animals 2025, 15(12), 1695; https://doi.org/10.3390/ani15121695 (registering DOI) - 8 Jun 2025

Abstract

Rodents are recognized as reservoirs of a wide range of pathogens, including microsporidia. The presence of microsporidia in the environment of mainland Spain and its islands has become increasingly known, as the number of studies has multiplied over time. The present study was [...] Read more.

Rodents are recognized as reservoirs of a wide range of pathogens, including microsporidia. The presence of microsporidia in the environment of mainland Spain and its islands has become increasingly known, as the number of studies has multiplied over time. The present study was conducted to determine the occurrence and diversity of microsporidia in three rodent species (Mus musculus, Rattus norvegicus, and Rattus rattus) in the Canary Islands, Spain. Ninety-three fecal samples were obtained from wild rodents on La Gomera and Gran Canaria Islands. Each sample was tested using Weber’s modified trichrome staining and immunofluorescence antibody tests (IFATs) against the Encephalitozoon genus and Enterocytozoon bieneusi. The microscopy-positive samples were subsequently analyzed using a nested polymerase chain reaction (PCR) followed by Sanger sequencing. The staining technique showed 38.7% (36/93) positivity, whereas the IFATs for Encephalitozoon spp. and Ent. bieneusi revealed 3.2% (3/93) and 6.5% (6/93) positivity, respectively. Finally, the nested PCR and nucleotide sequence analysis confirmed a 9.7% (9/93) occurrence of Ent. bieneusi and 17.2% occurrence (16/93) of different undetermined microsporidia species, whereas no Encephalitozoon spp. were detected. Seven different Ent. bieneusi genotypes were detected as follows: three known (AAE1, D, and SBM1) and four novel (GRE1, GRE2, LGE1, and LGE2), all of which belonged to Group 1. The results demonstrate, for the first time, that microsporidia are present in the rodent populations of the Canary Islands. Further studies are needed to determine the impact of the presence of microsporidia in rodents on the zoonotic transmission of these parasites. Full article

(This article belongs to the Section Mammals)

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13 pages, 947 KiB

Open AccessArticle

New Insights into the Phylogeographic History of Dirofilaria immitis in the Canary Islands, Spain

by Rodrigo Morchón, Alfonso Balmori-de la Puente, Manuel Collado-Cuadrado, Iván Rodríguez-Escolar, Noelia Costa-Rodríguez, Elena Infante González-Mohino, Elena Carretón and José Alberto Montoya-Alonso

Animals 2025, 15(12), 1694; https://doi.org/10.3390/ani15121694 (registering DOI) - 8 Jun 2025

Abstract

Heartworm disease (Dirofilaria immitis) is an important zoonotic infection of major clinical importance in dogs widespread, and transmitted by culicid vectors. Although D. immitis mostly affects dogs with an overall low incidence, some islands of the Atlantic archipelagos such as the [...] Read more.

Heartworm disease (Dirofilaria immitis) is an important zoonotic infection of major clinical importance in dogs widespread, and transmitted by culicid vectors. Although D. immitis mostly affects dogs with an overall low incidence, some islands of the Atlantic archipelagos such as the island of Gran Canaria (Canary Islands, Spain) had one of the highest historical prevalence/seroprevalence values in dogs, cats and humans. Molecular tools allow us to perform species identification diagnosis, phylogeographic and population genetics analysis that can provide key information about the factors making the disease still a threat (uncover untreated range of hosts, putative origin, etc.). In this study, we have optimized primers to amplify mitochondrial (COI, 12S) and nuclear (ITS) molecular markers from adult D. immitis worms. The genetic diversity and structure of D. immitis at the global level is limited, especially when compared with results obtained for other species of the same genus, such as D. repens. New minor haplotypes in the mitochondrial COI marker have been identified from adult D. immitis worms from infected dogs from the hyperendemic island of Gran Canaria, suggesting that the disease may have originated locally or may have been introduced from the mainland in historical times and evolved in isolation. To obtain a more complete understanding of its evolutionary history, structure and genomic diversity, comparative studies using next-generation sequencing data from endemic areas are needed, which will help in the long term to implement monitoring and control measures in a given area and to better understand its global phylogeographic history. Full article

(This article belongs to the Special Issue Vector-Borne and Zoonotic Diseases in Dogs and Cats: Second Edition)

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11 pages, 854 KiB

Open AccessArticle

Fecal Cortisol Metabolites Indicate Increased Stress Levels in Horses During Breaking-In: A Pilot Study

by Julia Krieber, Aurelia C. Nowak, Jakob Geissberger, Oliver Illichmann, Sabine Macho-Maschler, Rupert Palme and Franziska Dengler

Animals 2025, 15(12), 1693; https://doi.org/10.3390/ani15121693 (registering DOI) - 7 Jun 2025

Abstract

Sport horses are frequently exposed to situations that were identified as stressors, indicated by an increased cortisol release, which might impair animal welfare. However, while many studies deal with the impact of exercise, transport, and competition on stress in horses, little is known [...] Read more.

Sport horses are frequently exposed to situations that were identified as stressors, indicated by an increased cortisol release, which might impair animal welfare. However, while many studies deal with the impact of exercise, transport, and competition on stress in horses, little is known about the early phase of a horse’s sports career and studies investigating the stress level of young horses during breaking-in are limited. To compare stress levels in unridden horses, horses during breaking-in, and horses in training we collected fecal samples of young, unridden horses (n = 28) and of horses in different training stages (n = 13) and measured fecal cortisol metabolite (FCM) concentrations. Our preliminary results showed that FCM concentrations of unridden horses were significantly lower than those of horses in training (Mann–Whitney rank sum test, p < 0.001). Particularly in the first year under the saddle FCMs were significantly higher than in unridden horses (one way ANOVA + post hoc Holm–Sidak test, p < 0.05), with a tendency for FCM levels to decrease with time in training. Furthermore, we observed that within the group of ridden horses there was a larger range of variability in FCM levels, suggesting individual variations regarding their ability to deal with (training-induced) stress. These results indicate that breaking-in is a stressful time for young horses, underlining the importance of carrying out the initial training as carefully as possible. Full article

(This article belongs to the Special Issue Recent Advances in Equine Behavior and Welfare)

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11 pages, 259 KiB

Open AccessArticle

Effect of Guanidinoacetic Acid and Zilpaterol Hydrochloride Feed Additions on Lambs’ Productive Performance, Carcass Characteristics, and Blood Chemistry

by Daniel López-Aguirre, Javier Hernández-Meléndez, José F. Vázquez-Armijo, Luz Y. Peña-Avelino and Jorge Alva-Pérez

Animals 2025, 15(12), 1692; https://doi.org/10.3390/ani15121692 (registering DOI) - 7 Jun 2025

Abstract

This study evaluated the effects of dietary supplementation with guanidinoacetic acid (GAA) and zilpaterol hydrochloride (ZLH) on productive performance, carcass traits, and blood chemistry in non-castrated male lambs over 60 days. Twenty-four Pelibuey × Dorper crossbred lambs (16.3 ± 2.7 kg) were adapted [...] Read more.

This study evaluated the effects of dietary supplementation with guanidinoacetic acid (GAA) and zilpaterol hydrochloride (ZLH) on productive performance, carcass traits, and blood chemistry in non-castrated male lambs over 60 days. Twenty-four Pelibuey × Dorper crossbred lambs (16.3 ± 2.7 kg) were adapted to housing and diet for 14 days before being randomly assigned to one of three treatments: (1) Control: total mixed ration (TMR) without additives; (2) GAA: TMR with 0.2% GAA; and (3) ZLH: TMR with 6 mg/kg dry matter (DM) of ZLH for the last 30 days. No significant differences were observed in productive performance or carcass traits among treatments. However, lambs fed GAA showed higher serum glucose and creatinine levels than the control group (p < 0.05), suggesting a potential effect on energy metabolism. ZLH supplementation had no measurable impact on the parameters evaluated. These findings indicate that while GAA may influence certain metabolic indicators, further research with extended feeding periods or varying dosages is needed to clarify its effects on growth and carcass characteristics in lambs. Full article

(This article belongs to the Special Issue Sustainable Animal Production and Product Quality)

15 pages, 1891 KiB

Open AccessArticle

Effect of Pre-Freezing 18 °C Holding Time on Post-Thaw Motility and Morphometry of Cryopreserved Boar Epididymal Sperm

by Mamonene Angelinah Thema, Ntuthuko Raphael Mkhize, Maleke Dimpho Sebopela, Mahlatsana Ramaesela Ledwaba and Masindi Lottus Mphaphathi

Animals 2025, 15(12), 1691; https://doi.org/10.3390/ani15121691 (registering DOI) - 7 Jun 2025

Abstract

The study investigated the sperm motility and morphometry of pre-freeze and post-thaw boar epididymal semen cooled at increasing holding times at 18 °C. A total of 50 testes of heterogeneous boars were collected (5 testes/day) from the local abattoir and transported to the [...] Read more.

The study investigated the sperm motility and morphometry of pre-freeze and post-thaw boar epididymal semen cooled at increasing holding times at 18 °C. A total of 50 testes of heterogeneous boars were collected (5 testes/day) from the local abattoir and transported to the laboratory at 5 °C within 30 min after slaughter. Semen was retrieved from the caudal part of the epididymis using the slicing float-up method, diluted with Beltsville Thawing Solution extender, pooled in a 50 mL centrifuge tube/5 testes/day, and cooled at 18 °C. Following each holding time (0, 3, 6, 9, 12, 24, and 48 h), the cooled semen sample was re-suspended with Fraction A extender and stored at 5 °C for an additional 45 min. A cooled resuspended semen sample was then diluted with Fraction B extender, loaded into 0.25 mL straws, and frozen using liquid nitrogen vapour. Thawing was accomplished by immersing the semen straws in warm (37 °C) water for 1 min and the samples were evaluated for sperm motility and morphometry traits using the computer-assisted sperm analyzer system. The data were analyzed using variance analysis. Descriptive statistics were used to assess sperm morphometry, establishing the minimum and maximum values. Boar epididymal sperm survived for up to 48 h when held at 18 °C. Furthermore, the highest post-thawed sperm motility rates were observed in semen frozen after 3 h of holding time, with a sperm total motility of 85.9%, a progressive motility of 60.3%, and a rapid motility of 33.2%, as compared to other holding times (p < 0.05). The acceptable ranges for pre-freeze and post-thawed sperm morphology were head length (8.4–9.1 µm), width (4.4–4.8 µm), area (29.9–38.2 µm²), perimeter (20.1–23.7 µm), midpiece width (1.1–2.8 µm), and sperm shape, were consistent regardless of the holding time. A holding time of 3 h enhances the cryoresistance of sperm cooled at 18 °C. Therefore, these findings suggest that boar epididymal sperm can be effectively conserved and can maintain fertilization capability when cooled for 3 h at 18 °C before freezing. Full article

(This article belongs to the Section Animal Reproduction)

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12 pages, 636 KiB

Open AccessArticle

Cattle’s Social Rank Throughout the Transition from Rangeland to Fattening Affects Beef Quality

by Paola Soberanes-Oblea, Iván Adrián García-Galicia, Mariana Huerta-Jiménez, Jesús Ricardo Gámez-Piñón, Mieke Titulaer, Alma Delia Alarcon-Rojo and Einar Vargas-Bello-Pérez

Animals 2025, 15(12), 1690; https://doi.org/10.3390/ani15121690 (registering DOI) - 7 Jun 2025

Abstract

The social stress due to regrouping and hierarchy establishment in cattle in the transition from rangeland to feedlots may cause considerable losses in beef production due to aggressive interactions. This study determined the effect of the premortem social ranks of bovines in the [...] Read more.

The social stress due to regrouping and hierarchy establishment in cattle in the transition from rangeland to feedlots may cause considerable losses in beef production due to aggressive interactions. This study determined the effect of the premortem social ranks of bovines in the transition from rangelands to fattening pens on beef quality. The behavior of 20 heifers was evaluated in three premortem stages: 72 h after transportation, after 6 weeks of fattening, and while waiting at the slaughterhouse. Heifers were classified as dominant (D) or subordinate (S) based on ethograms. The principal component analysis revealed a strong relationship between social rank and beef color (L*, a*, b*, chroma, HUE, and ΔE), pH, and water holding capacity. The linear discriminant analysis of the beef quality variables classified the animals in one of four social ranks (dominant, change to subordinate, subordinate, change to dominant) with 95% precision. pH 45 (−9.163), drip 0 d (−3.917), final chroma (1.773), and ΔE (1.646) obtained high coefficients that determined the separation among the classes. Color variables like ΔE (p = 0.049) and chroma 0 d (p = 0.053) showed significant differences between the D and S animals in the MANOVA. Reactivity in dominants tended to negatively impact beef quality, affecting its color and pH. These findings contribute to a better understanding of how social dynamics impact beef quality during the transition period. Full article

(This article belongs to the Special Issue Animal Stress and Welfare During Transport and Slaughtering: Second Edition)

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12 pages, 611 KiB

Open AccessFeature PaperArticle

Cutaneous Allodynia of the Withers in Cattle: An Experimental In Vivo Neuroanatomical Preliminary Investigation of the Dichotomizing Sensory Neurons Projecting into the Reticulum and Skin of the Withers—A Case Study on Two Calves

by Roberto Chiocchetti, Luciano Pisoni, Monika Joechler, Adele Cancellieri, Fiorella Giancola, Giorgia Galiazzo, Giulia Salamanca, Rodrigo Zamith Cunha and Arcangelo Gentile

Animals 2025, 15(12), 1689; https://doi.org/10.3390/ani15121689 - 6 Jun 2025

Abstract

The presence of dichotomizing neurons in the dorsal root ganglia (DRG) of cattle, innervating both the reticulum and the withers, may indicate a pre-spinal convergence of visceral and cutaneous sensory information, i.e., that the DRG primary sensory neurons may elaborate the sensory information [...] Read more.

The presence of dichotomizing neurons in the dorsal root ganglia (DRG) of cattle, innervating both the reticulum and the withers, may indicate a pre-spinal convergence of visceral and cutaneous sensory information, i.e., that the DRG primary sensory neurons may elaborate the sensory information coming from two different anatomical areas before reaching the secondary sensory neurons within the spinal cord. This anatomical feature could be the underlying basis for the cutaneous allodynia observed in traumatic reticuloperitonitis, also known as the “Kalchschmidt pain test”. The aim of the study was to identify the DRG primary sensory neurons innervating the reticulum and the withers by using two different retrograde fluorescent tracers, Fast Blue (FB, affinity for cytoplasm) and Diamidino Yellow (DY, affinity for nucleus). In two anesthetized calves, FB and DY were injected into the reticulum and skin of the withers, respectively. At the end of the experimental period, the calves were deeply anesthetized and then euthanatized. The thoracic (T1–T8) DRG were collected and processed to obtain cryosections which were examined on a fluorescent microscope. A large number of neurons localized, especially in the T7 DRG, presented nuclei labeled with DY. On the contrary, only a few neurons localized exclusively in T6 and T7 DRG presented the cytoplasm labeled with FB. No neurons displayed FB and DY simultaneously within the cytoplasm and nucleus, respectively. The absence of double-labeled DRG neurons suggests that the convergence of visceral and somatic sensory inputs underlying the Kalchschmidt pain response likely does not occur at the level of individual DRG neurons. Rather, it may involve higher-order integrative centers, possibly including vagal pathways and brainstem nuclei which integrate the afferent information to coordinate respiratory movements of the diaphragm, intercostal muscles, and larynx. Although limited by the sample size, this case study provides a neuroanatomical basis for further investigation into central mechanisms of referred visceral pain in cattle. Full article

(This article belongs to the Section Cattle)

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12 pages, 731 KiB

Open AccessArticle

Effects of Heat Stress on Estrus Expression and Pregnancy in Dairy Cows

by Szilvia Szalai, Ákos Bodnár, Hedvig Fébel, Mikolt Bakony and Viktor Jurkovich

Animals 2025, 15(12), 1688; https://doi.org/10.3390/ani15121688 - 6 Jun 2025

Abstract

This study investigated how environmental temperature affects estrus behavior and pregnancy rates in lactating dairy cows during induced estrus. We hypothesized that higher temperatures reduce estrus expression and pregnancy rates and influence hormone levels. Fifty-eight healthy, multiparous, ovulation-synchronized cows were studied under heat [...] Read more.

This study investigated how environmental temperature affects estrus behavior and pregnancy rates in lactating dairy cows during induced estrus. We hypothesized that higher temperatures reduce estrus expression and pregnancy rates and influence hormone levels. Fifty-eight healthy, multiparous, ovulation-synchronized cows were studied under heat stress (summer) and non-stress (winter) conditions. Cows were assigned to G7G or OvSynch (OVS) hormone protocols. Blood samples collected on insemination day measured estradiol, LH, prolactin, insulin, and IGF-1 concentrations. Estrus signs and behaviors were also recorded. We found that during summer, daily maximum temperature–humidity Index (THI) values mostly exceeded 68, confirming the presence of heat stress. Estrus behaviors—especially standing and mounting—were significantly reduced under heat stress, occurring five to seven times more frequently in winter than in summer. Estrus detection rate was 86% in winter and 53% in summer. Pregnancy rates also declined sharply in summer (10%) compared to winter (39%), with all summer pregnancies in the G7G group. Hormonal analysis showed that estradiol levels remained consistent in the G7G group, while in the OVS group, it was significantly higher in winter than in summer. No significant seasonal differences were observed in the other hormones. Our findings indicate that heat stress impairs estrus expression and pregnancy rates in dairy cows. However, we conclude that the choice of hormone synchronization protocol (G7G) and the use of an automated estrus detection system may help to improve reproductive performance. Full article

(This article belongs to the Special Issue Heat Stress and Livestock: Effects on the Physiology)

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14 pages, 594 KiB

Open AccessArticle

Establishment and Validation of Sensitive Liquid Chromatography–Tandem Mass Spectrometry Method for Aldosterone Quantification in Feline Serum with Reference Interval Determination

by Tommaso Furlanello, Francesca Maria Bertolini, Andrea Zoia, Jose Sanchez del Pulgar and Riccardo Masti

Animals 2025, 15(12), 1687; https://doi.org/10.3390/ani15121687 - 6 Jun 2025

Abstract

Aldosterone, a mineralocorticoid hormone synthesised in the adrenal cortex, is essential for maintaining electrolyte balance and fluid homeostasis. Its role in feline physiology remains underexplored, despite its importance in regulating sodium reabsorption and potassium excretion via mineralocorticoid receptors in renal tubules. This study [...] Read more.

Aldosterone, a mineralocorticoid hormone synthesised in the adrenal cortex, is essential for maintaining electrolyte balance and fluid homeostasis. Its role in feline physiology remains underexplored, despite its importance in regulating sodium reabsorption and potassium excretion via mineralocorticoid receptors in renal tubules. This study is warranted given aldosterone’s importance in cats, particularly in light of their unique physiological traits, including highly concentrated urine and sensitivity to hydration status. Primary hyperaldosteronism, the most common feline adrenocortical disorder, contributes to arterial hypertension and chronic kidney disease, yet often remains underdiagnosed due to overlapping symptoms like hypokalaemia and hypertension. This research aimed to validate a liquid chromatography–tandem mass spectrometry (LC-MS/MS) method to measure serum aldosterone and to establish a reference interval in a population of healthy cats across a broad age range. The method demonstrated high precision and accuracy, with inter-assay coefficients of variation under 15%. Analysis of 49 healthy cats (40 young, 9 old) revealed a reference interval of 5.0–78.4 pg/mL (13.8–217.2 pmol/L). These findings provide a robust framework for diagnosing aldosterone-related disorders in cats and underscore the need for species-specific diagnostic tools. Improved understanding of aldosterone’s role could refine treatment strategies and enhance outcomes for affected feline patients. Full article

(This article belongs to the Special Issue Canine and Feline Endocrinology: Research Progress and Challenges)

16 pages, 1429 KiB

Open AccessArticle

Bioactive Element Biodistribution of Different Biological Substrates in Sheep and Goats

by Vincenzo Nava, Francesca Aragona, Angela Giorgia Potortì, Salvatore De Caro, Beatrice Di Bella, Federica Litrenta and Francesco Fazio

Animals 2025, 15(12), 1686; https://doi.org/10.3390/ani15121686 - 6 Jun 2025

Abstract

The production and reproduction of small ruminants, such as sheep and goats, living under extensive range-grazing conditions may be influenced by the availability of nutrients and the mineral content of forage. This study evaluated the biodistribution of calcium (Ca), copper (Cu), iron (Fe), [...] Read more.

The production and reproduction of small ruminants, such as sheep and goats, living under extensive range-grazing conditions may be influenced by the availability of nutrients and the mineral content of forage. This study evaluated the biodistribution of calcium (Ca), copper (Cu), iron (Fe), magnesium (Mg), manganese (Mn), sodium (Na) and zinc (Zn) in whole blood, serum, blood clots, plasma, plasma sediments and hair in sheep and goats. Forty clinically healthy, nonpregnant female sheep and goats were enrolled in the study. Hair and blood samples were performed in duplicate to assess mineral concentration in biological substrates using a Thermo Scientific iCAP- Q ICP-MS spectrometer (Omaha, Ne, USA). Higher levels of Ca, Cu, Fe, Mg, and Na were observed in blood and blood clots than in serum, plasma, plasma sediments and hair in sheep and goats. A significant effect of species was observed for all the bioactive element concentrations in the investigated substrates (p < 0.05). The results revealed positive correlations between serum and plasma for all investigated elements in sheep (p < 0.0001) and for Mn and Na in goats (p < 0.001), as well as between serum and blood for Ca, Fe, Mg and Mn in sheep (p < 0.0001) and for Ca and Mn in goats (p < 0.001). Plasma and blood showed a positive correlation for Ca, Fe, and Mn (p < 0.001) in sheep, and for Fe, Mn, Na, and Zn (p < 0.0001) in goats. These findings elucidate differences in biodistribution between sheep and goats, offering valuable insights for livestock production. Full article

(This article belongs to the Section Small Ruminants)

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10 pages, 455 KiB

Open AccessArticle

Molecular Prevalence and Genotyping of Toxoplasma gondii in Sheep Tissues Intended for Human Consumption in Shanxi Province, North China

by Xin-Sheng Lu, Jing Li, Chen Wang, Lu Wang, Xiao-Jing Wu, Xi-Long Yi, Ze-Xuan Wu, Wen-Bin Zheng and Xing-Quan Zhu

Animals 2025, 15(12), 1685; https://doi.org/10.3390/ani15121685 - 6 Jun 2025

Abstract

Toxoplasma gondii is one of the most widely distributed intracellular parasites worldwide, which can infect humans and a wide range of warm-blooded animals including sheep, with felines serving as its definitive host. T. gondii infection in sheep can lead to premature births, abortions [...] Read more.

Toxoplasma gondii is one of the most widely distributed intracellular parasites worldwide, which can infect humans and a wide range of warm-blooded animals including sheep, with felines serving as its definitive host. T. gondii infection in sheep can lead to premature births, abortions and stillbirths, causing significant economic losses to the sheep industry. Sheep farming has become a key pillar of the agricultural economy in Shanxi Province, North China, but little is known about T. gondii infection in sheep in this province. In the present study, a total of 755 sheep tissue samples (682 muscle tissue samples and 73 lymphatic tissue samples) were collected from different markets in 10 different cities of Shanxi Province. The genomic DNA of all samples was extracted and the B1 gene of T. gondii was amplified by PCR. The B1 gene-positive samples were genotyped at 12 genetic markers employing the multilocus nested PCR-restriction fragment length polymorphism (Mn-PCR-RFLP). The molecular prevalence of T. gondii infection in sheep tissues in Shanxi Province was 20.5% (155/755). The T. gondii genotype ToxoDB#9 was identified in one positive T. gondii sample, with complete genotyping at all 12 genetic markers based on Mn-PCR-RFLP. This is the first report of molecular prevalence and genotype of T. gondii infection in sheep in Shanxi Province. These results reveal the widespread distribution of T. gondii in sheep in Shanxi, which is of significant public health importance. Full article

(This article belongs to the Special Issue Coccidian Parasites: Epidemiology, Control and Prevention Strategies)

21 pages, 1248 KiB

Open AccessArticle

Mannan-Rich Fraction Supplementation: A Promising Nutritional Strategy for Optimizing Growth and Health of Pre-Weaning Calves

by Shanshan Guo, Yanfei Feng, Jianhao Yang, Haomiao Zhao, Jiajun Ma, Yuan Zhang, Mengkun Sun, Yifan Li, Gang Lin, Pengfei Lin, Aihua Wang and Yaping Jin

Animals 2025, 15(12), 1684; https://doi.org/10.3390/ani15121684 - 6 Jun 2025

Abstract

The early-stage feeding of calves significantly impacts their lifelong productive performance. During this period, the calves’ digestive tracts are in a dynamic state of development, making nutritional regulation a key factor in optimizing their growth potential. This research investigated the influence of mannan-rich [...] Read more.

The early-stage feeding of calves significantly impacts their lifelong productive performance. During this period, the calves’ digestive tracts are in a dynamic state of development, making nutritional regulation a key factor in optimizing their growth potential. This research investigated the influence of mannan-rich fraction (MRF) supplementation on pre-weaning calves. Sixty healthy Holstein calves were randomly divided into a control group and three MRF-supplemented groups (seven males and eight females per group) with low (L-MRF, 2.5 g/calf per day), medium (M-MRF, 5 g/calf per day), and high (H-MRF, 10 g/calf per day) doses. Growth performance, serum biochemical indicators, organ indices, intestinal morphology, and short-chain fatty acid (SCFA) concentrations were measured over a 42-day experiment. Compared with the control group, the M-MRF group had 4.9% higher overall body weight, body length, and starter intake, alongside a reduced diarrhea incidence by 4.6%. At day 14, MRF-supplemented groups exhibited significantly lower Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) levels, along with significantly higher Interleukin-12 (IL-12), growth hormone (GH), and Immunoglobulin G (IgG) levels. By day 42, IL-6 and Interleukin-1β (IL-1β) levels were also significantly decreased. Additionally, the lung and liver indices were higher and villus height was greater in both the jejunum and colon with MRF supplementation. In terms of SCFAs, the acetate level in the jejunum and the butyrate level in the colon were significantly higher in the MRF-supplemented calves. Both acetate in the jejunum and butyrate in the colon showed a significantly positive correlation with growth performance. Taken together, these findings suggest that MRF supplementation (particularly at 5 g/day) supports calf growth, gut health, and immunity, making it a promising nutritional strategy for pre-weaning calves. Full article

(This article belongs to the Section Cattle)

2 pages, 366 KiB

Open AccessCorrection

Correction: Luzardo et al. Integrating Conservation and Community Engagement in Free-Roaming Cat Management: A Case Study from a Natura 2000 Protected Area. Animals 2025, 15, 429

by Octavio P. Luzardo, Andrea Hansen, Beatriz Martín-Cruz, Ana Macías-Montes and María del Mar Travieso-Aja

Animals 2025, 15(12), 1683; https://doi.org/10.3390/ani15121683 - 6 Jun 2025

Abstract

In the original publication [...] Full article

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13 pages, 1842 KiB

Open AccessArticle

Bright-Field Multiplex Immunohistochemistry in Swine PCV2 and PRRSV Lymphadenopathies

by Giulia D’Annunzio, Luisa Vera Muscatello, Chiara Tugnoli, Stefano Pesaro, Andrea Luppi, Michelangelo Fiorentino, Tania Franceschini, Alessia Grillini, Gianluca Rugna, Giuseppe Sarli and Luciana Mandrioli

Animals 2025, 15(12), 1682; https://doi.org/10.3390/ani15121682 - 6 Jun 2025

Abstract

Multiplex immunostaining (mIHC) allows the simultaneous detection of multiple antigenic targets within the same tissue section, providing a deeper understanding of spatial variation in cellular distribution. The aim of the present study is to apply this technique to examine the spatial variation of [...] Read more.

Multiplex immunostaining (mIHC) allows the simultaneous detection of multiple antigenic targets within the same tissue section, providing a deeper understanding of spatial variation in cellular distribution. The aim of the present study is to apply this technique to examine the spatial variation of lymphocyte populations in swine lymph nodes during PCV2-SD and PRRSV lymphadenopathy compared with reactive lymphoid hyperplasia. A triple immunohistochemical stain with CD3, CD20 and IBA1 antibodies for the concurrent detection of T lymphocytes, B lymphocytes and macrophages, respectively, was performed. Multiplex immunohistochemistry (mIHC) revealed that, compared to reactive hyperplasia, the most significant changes in lymph node cell populations occurred in the follicles for both PCV2 and PRRSV infections. Additionally, in PCV2 cases, notable alterations were also observed in the interfollicular areas. In PCV2-affected lymph nodes, follicles not only significantly decreased in number but also showed a marked significant reduction in CD20⁺ and CD3⁺ cells. The interfollicular region in these cases also exhibited a significant reduction in CD3⁺ cells. In contrast, PRRSV-associated lymphadenopathy showed significantly increased CD20⁺ cells in the follicles, with a similar trend noted in the interfollicular region. mIHC provides more informative results on a single tissue section, thus preserving the topographical information of the tissue and allowing a comprehensive study of cellular composition, cellular functionality and cell–cell interactions, proving to be a valuable tool for studying and understanding disease dynamics. Full article

(This article belongs to the Special Issue Feature Papers in Veterinary Clinical Studies: From Current to Emerging Technologies and Innovative Approaches for the Future)

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26 pages, 10731 KiB

Open AccessArticle

Lactobacillus Supplementation Modulates Rumen Microbiota and Metabolism in Yaks Under Fattening Feeding Conditions: A Comprehensive Multi-Omics Analysis

by Jianlei Jia, Pengjia Bao, Ning Li, Siyuan Kong, Min Chu, Qian Chen and Ping Yan

Animals 2025, 15(12), 1681; https://doi.org/10.3390/ani15121681 - 6 Jun 2025

Abstract

The rumen is a critical organ that facilitates nutrient digestion in ruminant animals. However, the biological mechanisms by which rumen microbiota and its metabolites enable Lactobacillus to modulate rumen structure and maintain functional homeostasis under fattening feeding conditions remain poorly understood. In this [...] Read more.

The rumen is a critical organ that facilitates nutrient digestion in ruminant animals. However, the biological mechanisms by which rumen microbiota and its metabolites enable Lactobacillus to modulate rumen structure and maintain functional homeostasis under fattening feeding conditions remain poorly understood. In this study, 80 male Pamir yaks were selected, and a 170-day data collection phase was implemented. Correlation phenotypic data and multi-omics analyses (rumen microbial sequencing and rumen epithelial metabolomics) were conducted to investigate the regulatory effects of Lactobacillus supplementation on rumen microbiota and metabolic processes in a concentrate-based rearing yak model. The results demonstrated that feeding a high-energy diet may impair yak ruminal histomorphology, microbiota composition, and function while negatively modulating rumen microbiota–metabolic profiles associated with specific ruminal microbial communities and functions. Lactobacillus intervention treatment optimized the yak ruminal microbiome composition (mucous layer maturation was promoted, Prevotella and Ruminococcus abundance were reduced, and Fibrobacter and Muribaculaceae abundance were increased), thereby altering metabolite concentrations involved in various metabolic pathways under a high-energy feeding pattern (fatty acid metabolism pathways were upregulated). These alterations elucidated the beneficial impacts of the Lactobacillus supplementation strategy on yak ruminal health without compromising the high-energy intensive rearing pattern. Furthermore, the regulated ruminal microbiome metabolites may serve as potential biomarkers for future investigations into the functional impacts of Lactobacillus intervention treatment on healthy feeding strategies for yaks. Full article

(This article belongs to the Section Animal Nutrition)

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17 pages, 1111 KiB

Open AccessArticle

Effect of Cryodiluent and Time of Glycerol Addition on Cryopreservation and In Vitro Fertilization of Domestic Cat Epididymal Spermatozoa

by Natalia Gañán, Raquel González, Ana Sanchez-Rodriguez and Eduardo R. S. Roldan

Animals 2025, 15(12), 1680; https://doi.org/10.3390/ani15121680 - 6 Jun 2025

Abstract

Sperm cryopreservation and assisted reproduction are powerful tools for the conservation of endangered species. The domestic cat has been a useful model for studying wild felid reproductive biology due to the limited availability of endangered individuals for experimental research. Here, we investigate the [...] Read more.

Sperm cryopreservation and assisted reproduction are powerful tools for the conservation of endangered species. The domestic cat has been a useful model for studying wild felid reproductive biology due to the limited availability of endangered individuals for experimental research. Here, we investigate the effect of cryodiluents (TEST vs. Biladyl) and the timing of glycerol addition (before vs. after refrigeration, in one vs. three steps, respectively) on post-thaw sperm quality (motility, acrosome integrity) and their subsequent in vitro fertilization (IVF) ability with homologous oocytes. The results showed no statistically significant differences in sperm traits when samples were cryopreserved in TEST or Biladyl, or when glycerol was added in one or three steps. Motile sperm and intact acrosomes were significantly correlated before and after cryopreservation, indicating consistent relationships in fresh and thawed samples. The use of Biladyl significantly reduced IVF rates after cryopreservation compared to fresh sperm. Cryopreservation in TEST led to IVF rates that were not significantly different from those of fresh sperm. Using swim-up after thawing, or adding 1 mM pentoxifylline, did not enhance IVF results. Overall, a TEST cryodiluent with 4% glycerol added in one step is a reliable option for preserving epididymal cat spermatozoa. Full article

(This article belongs to the Special Issue Conservation and Sperm Quality in Domestic Animals)

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14 pages, 1632 KiB

Open AccessArticle

Innovation in the Processing of Native Round Fish: A Readjustment of the Processing Workflow for Salmonella spp. Control in a Fish Processing Plant in the State of Mato Grosso

by Jaqueline Oliveira Reis, Nathaly Barros Nunes, Yuri Duarte Porto, Adelino Cunha Neto, Sara Rodrigues de Souza, Washington da Guia Fonseca, Alexsandro Siqueira da Silva, Luciana Kimie Savay-da-Silva and Eduardo Eustáquio de Souza Figueiredo

Animals 2025, 15(12), 1679; https://doi.org/10.3390/ani15121679 - 6 Jun 2025

Abstract

Salmonella spp. is a pathogen detected in fish, although it is not part of its microbiota; the production and processing environment is the main source of contamination. Brazilian legislation recommends 5 ppm of free residual chlorine for fish washing, but Salmonella can still [...] Read more.

Salmonella spp. is a pathogen detected in fish, although it is not part of its microbiota; the production and processing environment is the main source of contamination. Brazilian legislation recommends 5 ppm of free residual chlorine for fish washing, but Salmonella can still be present. The objective of this study was to evaluate flaws in the processing flowchart and propose adjustments to reduce Salmonella spp. on the fish surface. Ninety samples were analyzed in a fish processing plant in Mato Grosso, divided into three treatments: (1) conventional processing, (2) modified flowchart, and (3) modified flowchart adapted to the specific plant conditions. Treatment 2 completely eliminated Salmonella spp., while treatment 3 reduced contamination to 3.3%, compared to 56.7% in conventional processing. The success of the modified treatment was only possible due to the main changes implemented in the process, which included the separation of dirty areas (responsible for gill and scale removal) and clean areas (designated for the careful removal of viscera without rupture and for filleting). No statistical difference was found between treatments 2 and 3 (p = 1, CI 0.00000–39.00055), suggesting that the adjusted flowchart can be implemented on a large scale to ensure food safety (OR = ∞, CI = [7.655, ∞], p < 0.001). This study highlights the effectiveness of the adjusted flowchart in reducing Salmonella spp. contamination in fish, with treatment 2 resulting in a complete absence of contamination. Treatment 3 maintained low contamination levels, demonstrating practical applicability in meatpacking plants. Full article

(This article belongs to the Section Animal Products)

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13 pages, 1825 KiB

Open AccessArticle

The First Report of Rhino DNA in Thailand: A Possible Extinct Indian Javan Subspecies, Rhinoceros sondaicus inermis

by Yada Katanyuphan, Pipad Krajaejun, Athiwat Wattanapituksakul and Wunrada Surat

Animals 2025, 15(12), 1678; https://doi.org/10.3390/ani15121678 - 6 Jun 2025

Abstract

The Javan rhino is a critically endangered species that has become extinct in several Asian countries, including Thailand. There are three Javan rhino subspecies: Rhinoceros sondaicus sondaicus, R. s. annamiticus, and R. s. inermis, and the first two subspecies existed [...] Read more.

The Javan rhino is a critically endangered species that has become extinct in several Asian countries, including Thailand. There are three Javan rhino subspecies: Rhinoceros sondaicus sondaicus, R. s. annamiticus, and R. s. inermis, and the first two subspecies existed in Thailand decades ago. However, no genetic study on Thai rhinos has been reported. In this study, we amplified a partial D-loop region of two ancient rhino specimens dated to approximately 100 years before present, from southwest Thailand. Then, we constructed phylogenetic trees and a haplotype network based on the D-loop sequences of the Thai rhinos and 39 Javan, Sumatran, and Indian rhinos from the GenBank database. The ancient Thai rhino DNA belonged to Javan rhinos and was clearly separated from R. s. annamiticus and R. s. sondaicus. The nucleotide differences among the ancient Thai, R. s. annamiticus, and R. s. sondaicus rhino DNA were 5.14–8.30%, similar to those between two white rhino subspecies. Moreover, species delimitation analyses confirmed that the ancient Thai rhinos should belong to another Javan rhino subspecies, possibly the extinct Indian Javan rhino. This is the first report of rhino DNA from Thailand and possibly the Indian Javan rhino DNA, which could be used for evolutionary studies. Full article

(This article belongs to the Section Mammals)

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