BMC Veterinary Research

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Virus sequences included in this study and their respective hosts
Article
Background Coronaviruses have the potential to cross species barriers. To learn the molecular intersections among the most common coronaviruses of domestic and close-contact animals, we analyzed representative coronavirus genera infecting mouse, rat, rabbit, dog, cat, cattle, white-tailed deer, swine, ferret, mink, alpaca, Rhinolophus bat, dolphin, whale, chicken, duck and turkey hosts; reference or complete genome sequences were available for most of these coronavirus genera. Protein sequence alignments and phylogenetic trees were built for the spike (S), envelope (E), membrane (M) and nucleocapsid (N) proteins. The host receptors and enzymes aminopeptidase N (APN), angiotensin converting enzyme 2 (ACE2), sialic acid synthase (SAS), transmembrane serine protease 2 (TMPRSS2), dipeptidyl peptidase 4 (DPP4), cathepsin L (and its analogs) and furin were also compared. Results Overall, the S, E, M, and N proteins segregated according to their viral genera (α, β, or γ), but the S proteins of alphacoronaviruses lacked conservation of phylogeny. Interestingly, the unique polybasic furin cleavage motif found in severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) but not in severe acute respiratory syndrome coronavirus (SARS-CoV) or Middle East respiratory syndrome coronavirus (MERS-CoV) exists in several β-coronaviruses and a few α- or γ-coronaviruses. Receptors and enzymes retained host species-dependent relationships with one another. Among the hosts, critical ACE2 residues essential for SARS-CoV-2 spike protein binding were most conserved in white-tailed deer and cattle . Conclusion The polybasic furin cleavage motif found in several β- and other coronaviruses of animals points to the existence of an intermediate host for SARS-CoV-2, and it also offers a counternarrative to the theory of a laboratory-engineered virus. Generally, the S proteins of coronaviruses show crossovers of phylogenies indicative of recombination events. Additionally, the consistency in the segregation of viral proteins of the MERS-like coronavirus (NC_034440.1) from pipistrelle bat supports its classification as a β-coronavirus. Finally, similarities in host enzymes and receptors did not always explain natural cross-infections. More studies are therefore needed to identify factors that determine the cross-species infectivity of coronaviruses.
 
Protein length and Genbank accession numbers of IRF4 in different species
Article
Background The interferon (IFN) regulatory factors (IRFs) were originally identified as transcription factors playing critical roles in the regulation of IFN-related genes in the signal pathway. In mammals, IRF4 plays a vital role in both the innate and adaptive immune system. This study aims to reveal the molecular characterization, phylogenetic analysis, expression profiles and the regulatory role in the IFN and NF-κB signalling pathways of IRF4 in common carp ( Cyprinus carpio . L) (abbreviation, ccIRF4). Results Here, ccIRF4 was identified and characterized, it contained a DNA binding domain (DBD) which possess five tryptophans and an IRF-associated domain (IAD). The predicted protein sequence of the ccIRF4 showed higher identities with grass carp ( Ctenopharyngodon idella ) and zebrafish ( Danio rerio ). Phylogenetic analysis suggested that ccIRF4 has the closest relationship with zebrafish IRF4. Quantitative real-time PCR analysis showed that ccIRF4 was constitutively expressed in all investigated tissues with the highest expression level in the gonad. Polyinosinic:polycytidylic acid (poly I:C) stimulation up-regulated the ccIRF4 expressions in the liver, spleen, head kidney, skin, foregut and hindgut. Upon Aeromonas hydrophila injection, the expression level of ccIRF4 was up-regulated in all tissues with the exception of spleen. In addition, ccIRF4 was induced by lipopolysaccharide (LPS), peptidoglycan (PGN) and Flagellin in head kidney leukocytes (HKLs). Overexpression of the ccIRF4 gene in epithelioma papulosum cyprini cells (EPC) down regulated the expressions of IFN-related genes and proinflammatory factors. Dual-luciferase reporter assay revealed that ccIRF4 decreased the activation of NF-κB through MyD88. Conclusions These results indicate that ccIRF4 participates in both antiviral and antibacterial immune response and negatively regulates the IFN and NF-κB response. Overall, our study on ccIRF4 provides more new insights into the innate immune system of common carp as well as a theoretical basis for investigating the pathogenesis and prevention of fish disease.
 
Article
  • Ramy E El-AnsaryRamy E El-Ansary
  • Wahid H El-DabaeWahid H El-Dabae
  • Ahmed S BreamAhmed S Bream
  • [...]
  • Ramy MohamedRamy Mohamed
Background: Lumpy skin disease (LSD), a disease of cattle and buffaloes, has recently become widely prevalent in Egypt. The aim of this study was to ascertain the potential role of Rhipicephalus (Boophilus) annulatus ticks in the transmission of this disease. Samples collected from suspected lumpy skin disease virus (LSDV) infected cows that had previously been vaccinated with the Romanian sheep pox virus (SPPV) in various Egyptian governorates were obtained between May to November over two consecutive years, namely 2018 and 2019. Ticks were morphologically identified and the partial cytochrome oxidase subunit I gene (COI) were sequenced, revealing that they were closely related to R. (Boophilus) annulatus. The G-protein-coupled chemokine receptor (GPCR) gene of the LSDV was used to test hard ticks. Results: Two positive samples from Kafr El-Sheikh province and one positive sample from Al-Behera province were reported. BLAST analysis revealed that the positive samples were closely related to the Kazakhstani Kubash/KAZ/16 strain (accession number MN642592). Phylogenetic analysis revealed that the GPCR gene of the LSDV recently circulating in Egypt belongs to a global cluster of field LSDV with a nucleotide identity of 98-100%. LSDV isolation was successfully performed four days after inoculation using 9 to 11-day-old embryonated chicken eggs showing characteristic focal white pock lesions dispersed on the choriallantoic membrane after three blind passages. Intracy-toplasmic inclusion bodies, cell rupture, vacuoles in cells, and virus particles ovoid in shape were demonstrated by electron microscopy. Conclusion: In this study the role of hard ticks in the transmission of the LSDV to susceptible animals in Egypt was revealed and confirmed by various methods.
 
Effects of SG on the growth performance of piglets. The ADG (A), ADFI (B), F/G (C) and DR (D) on days 0-10 (before infection); The DR (E) on days 13-14 (post infection). CP: Control group; STa: STa group; SG: SG + STa group. Values are means and SD, n = 8. a, b, Values within a column not sharing a common superscript letter indicate significant difference at P < 0.05
Effects of SG on blood biochemical and hematological parameters in pigs. The blood biochemical parameters (A) and hematological parameters (B) on day 10 (before infection); blood biochemical parameters (C) and hematological parameters (D) on day 15 (post infection). CP: Control group; STa: STa group; SG: SG + STa group. Values are mean and SD, n = 8. a, b, Values within a column not sharing a common superscript letter indicate significant difference at P < 0.05
Effects of SG supplementation on intestinal morphology of pigs. CP: Control group; STa: STa group; SG: SG + STa group. Values are mean and SD, n = 8. a, b, c, Values within a column not sharing a common superscript letter indicate significant difference at P < 0.05
Effects of SG on intestinal redox status in piglets. CP: Control group; STa: STa group; SG: SG + STa group. Values are mean and SD, n = 8. a, b, Values within a column not sharing a common superscript letter indicate significant difference at P < 0.05
Effects of dietary SG supplementation on colon bacteria of pigs after recombinant E. coli challenge. CP: Control group; STa: STa group; SG: SG + STa group. Values are mean and SD, n = 8. a, b, c, Values within a column not sharing a common superscript letter indicate significant difference at P < 0.05
Article
Background: The purpose of this research is to determine the effects of sodium gluconate (SG) on the growth performance and intestinal function in weaned pigs challenged with a recombinant Escherichia coli strain expressing heat-stable type I toxin (STa). Results: Pigs (n = 24, 21 days of age) were randomly allocated to three treatments: Control group (pigs were fed basal diet), STa group (pigs were fed basal diet and challenged with a recombinant E. coli strain expressing STa), and SG group (pigs were fed basal diet supplemented with 2500 mg/kg sodium gluconate and challenged with a recombinant E. coli strain expressing STa). The trial period lasted for 15 days. On days 12 and 13, pigs in the STa and SG groups were orally administered with the recombinant Escherichia coli strain, while those in the control group were orally administered with normal saline at the same volume. On day 15, blood, intestinal tissues and colonic contents were collected for further analysis. Results showed that dietary SG supplementation had a tendency to increase average daily gain, and reduced (P < 0.05) feed to gain ratio, plasma glucose concentration, and mean corpuscular hemoglobin concentration as compared with control group on days 0-10 of trial. Additionally, dietary SG supplementation attenuated(P < 0.05) the morphological abnormalities of small intestinal and the increase of the number of eosinophils in blood of pigs challenged with the recombinant Escherichia coli strain on day 15 of trial. Compared with control group, diarrhea rate and the number of eosinophils in blood and the concentrations of malondialdehyde in the jejunum were increased (P < 0.05). The height, width and surface area of the villi of the duodenum, the width and surface area of villi of jejunum and the height and width of villi of ileum were decreased (P < 0.05) in pigs challenged with the recombinant Escherichia coli strain in the STa group compared with those in control group on day 15 of trial. However, these adverse effects were ameliorated (P < 0.05) by SG supplementation in the SG group on day 15 of trial. Furthermore, dietary SG supplementation could reduce (P < 0.05) the total bacterial abundance in the colon, but SG did not restore the recombinant Escherichia coli-induced microbiota imbalance in colon. Conclusions: In conclusion, dietary supplementation with SG could improve piglet growth performance and alleviate the recombinant Escherichia coli-induced intestinal injury, suggesting that SG may be a promising feed additive for swine.
 
Article
Background Lumpy skin disease (LSD), a disease of cattle and buffaloes, has recently become widely prevalent in Egypt. The aim of this study was to ascertain the potential role of Rhipicephalus (Boophilus) annulatus ticks in the transmission of this disease. Samples collected from suspected lumpy skin disease virus (LSDV) infected cows that had previously been vaccinated with the Romanian sheep pox virus (SPPV) in various Egyptian governorates were obtained between May to November over two consecutive years, namely 2018 and 2019. Ticks were morphologically identified and the partial cytochrome oxidase subunit I gene (COI) were sequenced, revealing that they were closely related to R. (Boophilus) annulatus. The G-protein-coupled chemokine receptor (GPCR) gene of the LSDV was used to test hard ticks. Results Two positive samples from Kafr El-Sheikh province and one positive sample from Al-Behera province were reported. BLAST analysis revealed that the positive samples were closely related to the Kazakhstani Kubash/KAZ/16 strain (accession number MN642592). Phylogenetic analysis revealed that the GPCR gene of the LSDV recently circulating in Egypt belongs to a global cluster of field LSDV with a nucleotide identity of 98–100%. LSDV isolation was successfully performed four days after inoculation using 9 to 11-day-old embryonated chicken eggs showing characteristic focal white pock lesions dispersed on the choriallantoic membrane after three blind passages. Intracytoplasmic inclusion bodies, cell rupture, vacuoles in cells, and virus particles ovoid in shape were demonstrated by electron microscopy. Conclusion In this study the role of hard ticks in the transmission of the LSDV to susceptible animals in Egypt was revealed and confirmed by various methods.
 
Photomicrographs of jejunal tissue samples showing immunolabelling for Foxp3⁺ T cells, from control (a-b) and infected animals (c-h) showing different types of lesions associated with Map infection. (a-b). Positively immunolabelled cells show a bright brown stain in their cytoplasm. Control (C) animals with low presence of Foxp3⁺ T cells both in gut-associated lymphoid tissue (GALT) (a) and intestinal mucosa (MUCOSA) (b). (c-d) Animals with focal (F) lesions with a pronounced increase of immunolabelled Foxp3⁺ T lymphocytes is observed in the GALT (c) and to a lesser extent in the lamina propria (d). (e-f) Diffuse paucibacillary (PB) lesions where Foxp3⁺ T lymphocytes are also seen among the lymphocytic infiltrate of the LP (f) and in higher numbers on the GALT (e). (g-h) Diffuse multibacillary (MB) lesions with reduced presence of Foxp3⁺ T cells either in the GALT (g) or among the granulomas of the lamina propria (h), in similar amounts that the control (C) group. No specific distribution patter of the Foxp3⁺ T cells were detected in relation with the granulomas in any intestinal areas analyzed. Scale bar = 200 μm. Harris´s haematoxylin counterstain. e: epithelium lining the intestinal mucosa; lp: lamina propria; g: intestinal glands.
Bar-plot that shows the total mean cell counts and standard error (log-transformed) of positively immunolabelled Foxp3⁺ T cells subset according to the type of lesion. Different superscript letters indicate statistical significance between the different type of lesion. ***p < 0.001; *p < 0.05. Specific p-value resulting from each pairwise comparison between lesion category groups can be consulted in the Table 2.
Bar-plot that shows the mean cell counts and standard errors (log-transformed) of positively immunolabelled Foxp3⁺ T cell subset in the different intestinal areas analyzed according to the type of lesion. LP: Lamina propria; GALT: gut-associated lymphoid tissue; MLN: mesenteric lymph node. Different superscript letters indicate statistical significance between the different intestinal areas within each type of lesion. **p < 0.01; *p < 0.05.Specific p-value resulting from each pairwise comparison between intestinal areas within each type of lesion can be consulted in the Table 2.
Article
Background Mycobacterium avium subsp. paratuberculosis infected animals show a variety of granulomatous lesions, from focal forms with well-demarcated granulomas restricted to the gut-associated lymphoid tissue (GALT), that are seen in the initial phases or latency stages, to a diffuse granulomatous enteritis, with abundant (multibacillary) or scant (paucibacillary) bacteria, seen in clinical stages. Factors that determine the response to the infection, responsible for the occurrence of the different types of lesion, are still not fully determined. It has been seen that regulatory T cells (Treg) play an important role in various diseases where they act on the limitation of the immunopathology associated with the immune response. In the case of paratuberculosis (PTB) the role of Treg lymphocytes in the immunity against Map is far away to be completely understood; therefore, several studies addressing this subject have appeared recently. The aim of this work was to assess, by immunohistochemical methods, the presence of Foxp3 ⁺ T lymphocytes in intestinal samples with different types of lesions seen in cows with PTB. Methods Intestinal samples of twenty cows showing the different pathological forms of PTB were evaluated: uninfected controls ( n = 5), focal lesions ( n = 5), diffuse paucibacillary ( n = 5) and diffuse multibacillary ( n = 5) forms. Foxp3 ⁺ lymphocyte distribution was assessed by differential cell count in intestinal lamina propria (LP), gut-associated lymphoid tissue (GALT) and mesenteric lymph node (MLN). Results A significant increase in the number of Foxp3 ⁺ T cells was observed in infected animals with respect to control group, regardless of the type of lesion. However, when the different categories of lesion were analyzed independently, all individuals with PTB lesions showed an increase in the amount of Foxp3 ⁺ T lymphocytes compared to the control group but this increase was only significant in cows with focal lesions and, to a lesser extent, in animals with diffuse paucibacillary forms. The former showed the highest numbers, significantly different from those found in cows with diffuse lesions, where no differences were noted between the two forms. No specific distribution pattern was observed within the granulomatous lesions in any of the groups. Conclusions The increase of Foxp3 ⁺ T cells in focal forms, that have been associated with latency or resistance to infection, suggest an anti-inflammatory action of these cells at these stages, helping to prevent exacerbation of the inflammatory response, as occurs in diffuse forms, responsible for the appearance of clinical signs.
 
The proportion of patients allocated to each healing category, in open and closed repair groups
Comparing the number of repeat visits for open and closed repair techniques
Comparing the number of complications in the open and the closed repair group
Comparing the number of cases experienced malalignment between open and close groups
Article
Backfround Treatment options for metacarpal/metatarsal fractures include conservative and surgical management. The aim of this study is to determine whether there is any significant difference in healing and complication rates, between open and closed treatment. Medical records of dogs and cats with metacarpal/metatarsal fractures with complete follow-up were retrospectively reviewed. Patients were allocated in two groups: open or closed stabilization. Minor and major complications were recorded and compared. Fracture healing was classified as good, delayed and non-union, and it was statistically compared. Results Sixty-three patients (35 dogs and 28 cats) were included. Thirty-one were treated with an open approach and 32 by a closed stabilization. Regarding fracture healing a significantly higher proportion of delayed healing/non-union was found in the closed group (12/32 vs 2/31). Regarding postoperative complications, a significantly higher number of animals in the open group did not develop any complications (12/31 vs 3/32). A significantly higher proportion of minor complications were reported in the closed group (27/32 vs 12/31). However, a higher number of major complications was reported in the open group (7/31 vs 2/32) although this was not statistically significant. Fracture malalignment was significantly more prevalent in patients undergoing closed stabilization (11/32 vs 2/31). Conclusion According to the results, better healing, fracture alignment and a lower complication rate are found when fractures are stabilised with an open technique. However, other factors such as configuration of the fracture, soft tissue involvement, patient´s character and client´s situation would also need to be taken into account in the decision of stabilization technique.
 
Article
Abstract Understanding the does reproductive hemodynamic changes during the estrous cycle is crucial for improving reproductive competence and fertility potential in this species. The objective of this study is to investigate the hemodynamic variations in ovarian (OA) and uterine (UA) arteries, histological and morphometric changes in ovarian and uterine tissues throughout the follicular (FP) and luteal (LP) phases in rabbits and determine estrogen (ER), progesterone (PR) receptors, and vascular endothelial growth factor (VEGF) distributions using immunohistochemistry. Fourteen adults pluriparous New Zealand rabbits were divided into rabbits at the FP (Day − 1; n = 7) and those at the LP (Day 9; n = 7). Animals were subjected to Doppler, hormonal (estrogen [E2], progesterone [P4], insulin-like growth factor [ILGF], and VEGF), histological, and immunohistochemical analyses. In LP, OA Doppler indices were significantly increased, whereas peak systolic velocity (PSV) was decreased compared with that in FP. UA Doppler indices were significantly decreased in the LP, whereas PSV was increased (P
 
AP 300 radiant catalytic ionization device
Diagram of the RCI room with the location of all elements, including the RCI instrument, airflow (input and output) and sampling sites (M – microbiological, A – ammonia, D – dust)
CFU differentiation for total airborne bacteria and fungi between the control room and RCI room on the 1st and 7th days of the experiment, significantly different at *p < 0.05 and **p < 0.05, Kruskal–Wallis test
Bacterial and fungal CFUs in the bedding of cages in the control and RCI room on the 1st and 7th days of the experiment, **significant difference at p < 0.05, Kruskal–Wallis test
Effect of 7 days of RCI on ammonia gas concentrations in mouse rooms, * significantly different at **p < 0.01, Mann–Whitney U test
Article
High stocking densities, closed animal houses, and elevated concentrations of bacteria, fungi, and the products of their activity, including ammonia and hydrogen sulphide, have adverse health effects. Active techniques used to reduce unfavourable environmental conditions, such as ventilation, sprinkling, bedding sorbents, and nutritional treatments, are not always sufficient to improve the animals’ living environment. The current paper aims to evaluate the effect of radiant catalytic ionization (RCI) on airborne microorganisms, cage microbiological status, gaseous ammonia concentrations, and the haematological status of mice in animal houses. After one week of operation of an RCI system, the number of airborne bacteria and fungi in the experimental room decreased in comparison to the first day of the experiment ( p < 0.05 and p < 0.05 respectively), as did the concentrations of ammonia ( p < 0.01) and dust. At the same time, the basic health parameters of the mice, determined in the blood, were very similar between the control and experimental room. RCI seems to be an ideal solution to ensure high hygiene standards in animal rooms and houses with limited use of disinfectants or antibiotic treatment of sick animals. An additional, environmental benefit is the limited amount of nitrogen released.
 
Clinical signs and gross lesions of cattle affected by LSD. Distribution of skin nodules through the body surface of a calf (A), swelling of udder accompanied by mastitis (B), enlargement of prefemoral (C), and prescapular lymph node (D), corneal opacity or keratitis (E), and sit fast lesion (F)
Histopathology of LSD: H&E.Ballooning degeneration (arrow) and intracytoplasmic eosinophilic inclusion bodies (arrowhead) 100 X (a). Hydropic degeneration on the epidermis (arrow) and edema on the dermis (arrowhead) 10 X (b). Inset indicates higher magnification of a and b (40X) (c); Eosinophilic intracytoplasmic inclusion bodies on epidermal cells (arrowhead) (d). Characteristic eosinophilic intracytoplasmic inclusion bodies in follicular cells (arrowhead) (e); Fibrin necrotic vasculitis (f)
Amplification fluorescence curves
Article
Background Lumpy skin disease is a contagious viral disease of cattle caused by LSDV that results in huge economic losses in the cattle industry. This study characterizes LSDV in cattle through clinicopathological and molecular techniques in selected districts of Wolaita Zone, Southern Ethiopia. Methods A crossectional study was conducted from November 2020 to June 2021 using Real-time polymerase chain reaction and Histopathological techniques to confirm LSDV. Result This study revealed that the percentage of positivity of cattle for LSDV was 36.2%. Clinically, cattle infected with LSDV revealed fever (39–41 °C), nodular lesions on the skin and mucous membranes, and lymphadenopathy. Histopathologically, affected tissue revealed ballooning degenerations of the epidermis, infiltration of mononuclear inflammatory cells, vasculitis, and intracytoplasmic eosinophilic inclusion bodies. RT-PCR confirmed that DNA extracts from skin biopsies of virus isolates were positive for LSDV. Conclusion The present study confirms that LSDV is widely circulating in cattle of selected districts of the Wolaita zone. Thus, effective control measures through regular vaccination and further confirmation of circulating strains of LSDV through detailed molecular analysis should be recommended.
 
Flow of participants in the study. SDM-Q-9, Shared Decision Making Questionnaire – pet owner; SDM-Q-Doc, Shared Decision Making Questionnaire – veterinarian
Article
Abstract Background Communication skills are a necessary competency in veterinary medicine, and shared decision-making (SDM) between practitioners and patients is becoming increasingly important in veterinary practice as in human medicine. There are few studies that have quantitatively measured SDM in veterinary health care, and the relationship between SDM and consultation satisfaction is unknown. The purpose of this study was to investigate the status of SDM implementation in veterinary hospitals and the relationship between SDM implementation and consultation satisfaction among pet owners. We conducted a cross-sectional study using self-administered questionnaires among pet owners and veterinarians. In total, 77 pet owners who visited a veterinary clinic and 14 veterinarians at the clinics participated in this study. After a veterinary clinic visit, owners were asked to rate their decision-making preferences using the Shared Decision Making Questionnaire for patients (SDM-Q-9) adapted for veterinary medicine, as well as their satisfaction with the consultation. The corresponding veterinarians were asked to complete the veterinary version of the survey (SDM-Q-Doc). Results Most pet owners (64.9%) preferred SDM in veterinary consultations. Cronbach's alpha coefficient of 0.84 for the veterinary SDM-Q-9 and 0.89 for the veterinary SDM-Q-Doc both confirmed high reliability. The Spearman's correlation coefficient between the SDM-Q-9 and consultation satisfaction was 0.526 (p
 
Location of study area. Districts where outbreaks occurred are shown in green range (Xilingol, Inner Mongolia)
Phylogenetic tree with Neighbor-Joining (NJ) shows the relationship between LSDV full-length genomic sequences from LSDV/NMG/2020, marked with red round, with other Capripoxvirus gene sequences from GenBank
Phylogenetic trees with Neighbor-Joining (NJ) show the relationship between LSDV P32, RPO30 and EEV genomic sequences from LSDV/NMG/2020, marked with red round, with other Capripoxvirus gene sequences from GenBank
Phylogenetic trees with Neighbor-Joining (NJ) show the relationship between LSDV GPCR and LSDV117 genomic sequences from LSDV/NMG/2020, marked with red round, with other Capripoxvirus gene sequences from GenBank
Article
Background The outbreak of Lumpy skin disease (LSD) in cattle caused by LSD virus (LSDV) was first reported in August 2019 in China. Since then, several LSD outbreaks have been reported in seven different provinces of China. Until now, several Lumpy skin disease virus (LSDV) strains from China have been reported and sequenced including LSDV/Xinjiang/2019 (MN598005.1), China/GD01/2020 (MW355944.1), and LSDV/Hongkong/2021 (MW732649.1). In October 2020, more than 1,700 cattle imported from Chile arrived in Xilingol, Inner Mongolia, and were diagnosed with LSD. Currently, limited data on the origin of the virus is available. Methods Nucleotide sequences of the ORF11, ORF36, ORF74, ORF117, ORF126 genes and the complete genome of LSDV strains and isolates were downloaded from NCBI database. MEGA7.0 was used to perform phylogenetic analysis with Neighbor-Joining (NJ). DNASTAR software is used to analyze homologous comparison analysis with related genes of reference strains included in Genbank. Results Compared with other strains isolated from China, the results of full genome sequence analysis showed the LSDV/NMG/2020 strain belonged to the recombinant strains. The LSDV/NMG/2020 strain is different from the current LSDV field isolates in Africa, the Middle East, Europe, and the newly emerged LSDV Russia variants. Based on the identities of P32, RPO30, EEV, GPCR and LSDV117 genes (99.8%, 99%, 99.8%, 99% and 98.7%), the sub-cluster recombinant containing LSDV/NMG/2020 strain is phylogenetically closer to the Russia strain (Saratov/2017). Conclusions In this study, we reported a new isolated LSDV strain named LSDV/NMG/2020. The results of genomic characterization and phylogenetic analysis demonstrated that the LSDV/NMG/2020 isolate was a vaccine-like recombinant strain.
 
Temporal profile of plasma glucose (A), insulin (B) and c-peptide (C) and total GLP-1 (D) during the oral sugar test (OST). The dark shaded area (with mean represented with light grey line) indicates the 95% confidence interval of the average without treatment whilst the light shaded area indicates the 95% confidence interval for the average (dark grey line) with exenatide treatment
Spaghetti plots of changes with treatment per horse in area under the curve (AUC) of plasma glucose (A), peak plasma glucose (B), AUC plasma insulin (C), peak plasma insulin (D). The AUC of glucose was significantly decreased (P = 0.007, panel A) with treatment. Furthermore, the peak plasma glucose concentrations during the oral sugar test (OST) were also significantly reduced in the treatment group (P < 0.001; panel B). The AUC of insulin was significantly higher (P = 0.003; panel C) in the control period in comparison to the treatment. The peak insulin concentration (Cmax) during the OST was significantly reduced with treatment by 14.35 µU/ml (P = 0.003; panel D)
Bar graph of the marginal insulin sensitivity (SI) values for the placebo and exenatide treatment. Estimates of SI were significantly higher (P = 0.02) after treatment with exenatide (mean SI: 7.49 10–4· µU/ml⁻¹·min⁻¹; 95% CI: 3.46 – 11.52 10–4· µU/ml⁻¹·min⁻¹) in comparison to control (mean SI: 1.93 10–4· µU/ml⁻¹·min⁻¹; 95% CI: 0.005 – 3.86 10–4· µU/ml⁻¹·min⁻¹)
Article
Background Insulin dysregulation (ID) is the most important risk factor for the development of laminitis in horses and therapies to control it are needed. Hypothesis/objectives To assess the effects of a single dose of the synthetic GLP-1 analog exenatide on postprandial insulin dynamics. We hypothesized that exenatide would improve insulin sensitivity and lower postprandial blood insulin concentrations. Study design Randomized, crossover, experimental study. Animals Six horses (3 mares, 3 geldings; 2 with normal insulin regulation [NIR] and 4 with mild ID). Methods Horses completed both study arms: subcutaneous administration of exenatide (or no treatment) 30 min before an oral sugar test (0.15 ml/kg of Karo Syrup). Blood samples obtained over 240 min were assayed for glucose, insulin, lactate, c-peptide and total GLP-1. The area under the curve (AUC) was calculated using the trapezoidal rule. Insulin sensitivity ( S I ) was estimated using a mathematical model. Results Exenatide resulted in a postprandial decrease of 20% (effect size: 2673 µU·min/ml; 95% CI: 900 – 4446 µU·min/ml; P = 0.003) in AUC of plasma insulin (control; mean AUC insulin: 11,989 µU·min/ml; 95% CI: 9673 – 14,305 µU·min/ml, exenatide; mean AUC insulin: 9316 µU·min/ml; 95% CI: 7430 – 11,202 µU·min/ml). Exenatide resulted in an approximately threefold increase (effect size: 5.56 10 –4 · µU/ml ⁻¹ ·min ⁻¹ ; 95% CI: 0.95 – 10.1 10 –4 · µU/ml ⁻¹ ·min ⁻¹ ; P = 0.02) in estimated insulin sensitivity (control mean S I : 1.93 10 –4 · µU/ml ⁻¹ ·min ⁻¹ ; 95% CI: 0.005 – 3.86 10 –4 ·µU/ml ⁻¹ ·min ⁻¹ vs. exenatide mean S I : 7.49 10 –4 · µU/ml ⁻¹ ·min ⁻¹ ; 95% CI: 3.46 – 11.52 10 –4 · µU/ml ⁻¹ ·min ⁻¹ ). Conclusions The decrease in insulin response to carbohydrates was due to an increase in whole-body insulin sensitivity. GLP-1 agonists may have therapeutic potential for ID in horses.
 
Article
Staphylococcus aureus is a common mastitis pathogen in dairy cows, and methicillin-resistant S. aureus (MRSA) has been found in dairy farms all over the world. The study carried out on bovines from three governorates in Egypt, with the goal of determining the prevalence of MRSA in positive milk samples of subclinical mastitis, performing an antibiotic susceptibility test against MRSA isolates and determining the risk factors associated with MRSA. A total of 350 quarter milk samples (n = 200 mixed breed cow; n = 150 water buffalo) were collected and examined for subclini-cal mastitis using the California mastitis test (CMT) before being exposed to standard microbiological procedures for S. aureus isolation. The disc diffusion method was used to phenotypically analyse the positive S. aureus isolates for MRSA, which was verified by a PCR assay targeting the mecA gene. According to the findings of the study, 41.4% (145/350) milk samples were positive based on CMT, while 35.7% (125/350) of positive samples identified as MRSA based on PCR assay. However, the obtained results revealed non-significant disparity between cattle and buffalo and all predicted risk factors were strongly associated with prevalence of subclinical mastitis. The in-vitro antibiotic susceptibility test revealed that cefoxitin was completely resistant, whereas linezolid, ciprofloxacin, and trimethoprim + sulphamethoxa-zole were sensitive against the MRSA isolates. The relevance of S. aureus to public health, as well as the development of resistance to antibiotics like methicillin, needs ongoing testing of antimicrobial medications against MRSA isolates.
 
The present study was conducted in four counties of Kurdistan province, including Marivan, Baneh, Sanandaj, and Divandarreh. The location of these counties on the district map of Kurdistan province is indicated with a green star
Article
Background Q fever is one of the most important zoonotic diseases caused by Coxiella burnetii . Although Q fever is an endemic disease in Iran, epidemiological data on C. burnetii infection are not yet complete in reservoirs and vectors in some parts of Iran. This survey investigated C. burnetii infection in small ruminants (sheep and goat blood samples) and their ticks in western Iran (Kurdistan province) in 2020. The presence of C. burnetii DNA was identified in these samples by targeting the IS1111 gene using the quantitative PCR (qPCR) method. Results Out of 250 blood samples (232 sheep and 18 goats), C. burnetii was detected in two samples (0.8%) belonging to the sheep (0.9%). In addition, 34 of 244 collected ticks (13.9%) from infested animals (244) were positive for C. burnetii infection. The highest prevalence of infection was found in Dermacentor marginatus (18.3%) and Haemaphysalis concinna (12.5%). Conclusions The present study showed that ticks could have a possible role in the epidemiology of Q fever in Iran.
 
Schedule of ovine multiple ovulation. At the start of experiment, the estrus was detected and the ewes were divided into CIDR induced estrus (IE) and Spontaneous estrus (SE) group (65 and 21, respectively). The IE group was treated as upper: the CIDR was inserted into vagina of non-estrus ewes at random day (D0); at D10 to 12, the FSH was injected for 6 times; at first time of FSH injection, the PMSG was injected synchronously; at fifth time of FSH injection, the PG was injected synchronously; the CIDR was removed at last time of FSH injection; at D13, the estrus was detected and AI was performed twice (12 h interval); the LH was injected at first AI; at D19, the embryos were collected via surgical uterus flushing. The SE group was treated as under: the estrus day of ewes as D0; at D13 to 15, the FSH was injected for 6 times; at first time of FSH injection, at last time of FSH injection, the PG was injected synchronously; at D16, the estrus was detected and AI was performed twice (12 h interval); the LH was injected at first AI; at D22, the embryos were collected via surgical uterus flushing
The first time (1st) Metabolomic analysis of serum from ewes with multiple ovulation treatment. PCA analysis results of IE group under positive ion mode (a) and negative ion mode (b) and SE group under positive ion mode (c) and negative ion mode (d). PLS-DA analysis results of IE group under positive ion mode (c) and negative ion mode (d) and SE group under positive ion mode (g) and negative ion mode (h)
The second time (2nd) Metabolomic analysis of serum from ewes with multiple ovulation treatment. PCA analysis results of IE group under positive ion mode (a) and negative ion mode (b) and SE group under positive ion mode (c) and negative ion mode (d). PLS-DA analysis results of IE group under positive ion mode (c) and negative ion mode (d) and SE group under positive ion mode (g) and negative ion mode (h)
Volcano plot of first comparation analysis (1st) between high and low embryonic yield populations of serum from ewes with multiple ovulation treatment. IE group under positive ion mode (a) and negative ion mode (b) and SE group under positive ion mode (c) and negative ion mode (d)
Volcano plot of second comparation analysis (2nd) between high and low embryonic yield populations of serum from ewes with multiple ovulation treatment. IE group of positive ion mode (a) and negative ion mode (b) and SE group of positive ion mode (c) and negative ion mode (d)
Article
Background The establishment of non-invasive diagnostic method for multiple ovulation prediction is helpful to improve the efficiency of multiple ovulation. The blood hormones and metabolites would be suitable indexes for this subject. Methods In this study, 86 estrus ewes (65 of induced estrus (IE) and 21 of spontaneous estrus (SE)) were selected and the blood samples were collected at the day before follicle-stimulating hormone (FSH) injection (1 st ) and before artificial insemination (2 nd ). The serum reproductive hormones ofFSH, luteinizing hormone (LH), 17β-Estradiol (E2), progesterone (P4) and anti-Mullerian hormone (AMH) were measured through enzyme linked immunosorbent assay (ELISA) and the untargeted metabolomics analysis was processed through LC–MS/MS. The embryos were collected after 6.5 days of artificial insemination. Results In total, 975 and 406 embryos were collected in IE and SE group, respectively. The analysis of reproductive hormones showed that concentrations of FSH, E2 and AMH were positive correlated with the embryo yield while concentrations of LH and P4 were negative correlated in both group at 1 st detection. At 2 nd detection, the trends of reproductive hormones were similar with 1 st except P4, which was positive correlated with embryo yield. The metabolomics analysis showed that 1158 metabolites (721 in positive iron mode and 437 in negative iron mode) were detected and 617 were annotated. In 1 st comparation of high and low embryonic yield populations, 56 and 53 differential metabolites were identified in IE and SE group, respectively. The phosphatidyl choline (PC) (19:0/20:5) and PC (18:2/18:3) were shared in two groups. In 2 nd comparation, 48 and 49 differential metabolites were identified in IE and SE group, respectively. The PC (18:1/18:2) and pentadecanoic acid were shared. Most differential metabolites were significantly enriched in amino acid, fatty acid metabolism, digestive system secretion and ovarian steroidogenesis pathways. Conclusions This study showed that FSH, P4, AMH, the PC relevant metabolites and some anomic acids could be potential biomarkers for embryonic yield prediction in ovine multiple ovulation. The results would help to explain the relation between blood material and ovarian function and provide a theoretical basis for the multiple ovulation prediction.
 
FNA of the liver of a Kenyan sand boa (Eryx colubrinus loverdigei). Mixed inflammation. Multiple unstained rod-shaped bacteria are visible in the sample background (red arrows). Reactive lymphocytes (arrowhead) and partially degranulated heterophils (black arrow) were numerous. Diff Quick® stain, 400x. Scale bar: 10 μm\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\mathrm{\mu m}$$\end{document}
Kenyan sand boa (Eryx colubrinus loverdigei), necropsy. At the coelomic cavity opening, numerous miliary granulomas are visible adherents to serous membranes
Liver of the Kenyan sand boa (Eryx colubrinus loverdigei), gross pathology. Multiple miliary nodules are disseminated in all the liver parenchyma
A Liver Kenyan sand boa (Eryx colubrinus loverdigei), 10X, Hematoxylin and Eosin stain. Multiple granulomas with central necrosis (black arrows). B Liver, 10X, Fite-Faraco stain. Many aggregates of positive red bacilli within granuloma’s centers. Scale bar: 200 μm\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\mathrm{\mu m}$$\end{document}
Article
Background Environmental nontuberculous mycobacteria species that are not members of the M. tuberculosis complex, are ordinary inhabitants of a wide variety of environmental reservoirs and their role in human and animal diseases has been fully recognized. Even if spontaneous mycobacterial infections have been reported in a wide variety of reptiles, this is the first report of systemic fatal mycobacteriosis sustained by Mycobacterium goodii in a pet reptile. Case presentation An adult, wild caught (WC), male Kenyan sand boa ( Eryx colubrinus loveridgei ) age unknown, was presented for clinical examination due to decreased activity level, decreased appetite and diarrhea. Blood tests showed unreliable results. Coprologic exam showed a moderate to severe presence of flagellates. X rays and ultrasound showed moderate presence of air and faeces in the large intestine. The snake was hospitalized and oral metronidazole was chosen as antiprotozoal agent in association with subcutaneous warm fluids. The snake was discharged after 2 weeks therapy in good clinical condition. Faecal exam resulted negative. One month after, the snake was quickly hospitalized again because of a recrudescence of symptoms. Biochemistry showed severe increase of AST, ALT and biliary acids. Severe leucocytosis and moderate to severe anemia were highlighted. Ultrasound examination revealed a severe diffused alteration of the liver parenchyma and a fine needle aspiration was performed. The cytological diagnosis was mixed inflammation, with a numerous of unstained rod-shaped bacteria both inside macrophages and free in the sample. The snake’s condition rapidly deteriorated and euthanasia was performed. The histology of the coelomic organs confirmed a systemic mycobacteriosis. Real-time PCR identified the mycobacteria as Mycobacterium goodii . Conclusions Species from the genus Mycobacterium are among the most important micro-organism including the causative agents of tuberculosis. Even if the general incidence of disease in reptiles due to mycobacteria is comparatively low, they can serve as reservoirs of many ubiquitous mycobacteria species. Mycobacterium goodii is a rapidly growing non‐tuberculous mycobacterium that has recently been associated with severe infections in animals and humans. Although in this case the pathogenesis was not completely clear, we highlight the zoonotic risk of mycobacteriosis in exotic animals especially in WC specimens.
 
Article
Background This study investigated the effects of chronic heat stress on liver inflammatory injury and its potential mechanisms in broilers. Chickens were randomly assigned to the 1-week control group (Control 1), 1-week heat stress group (HS1), 2-week control group (Control 2), and a 2-week heat stress group (HS2) with 15 replicates per group. Broilers in the heat stress groups were exposed to heat stress (35 ± 2 °C) for 8 h/d for 7 or 14 consecutive days, and the rest of 26 hours/day were kept at 23 ± 2 °C like control group broilers. Growth performance and liver inflammatory injury were examined for the analysis of liver injury. Results The results showed that heat stress for 2 weeks decreased the growth performance, reduced the liver weight ( P < 0.05) and liver index ( P < 0.05), induced obvious bleeding and necrosis points. Liver histological changes found that the heat stress induced the liver infiltration of neutrophils and lymphocytes in broilers. Serum levels of AST and SOD were enhanced in HS1 ( P < 0.01, P < 0.05) and HS2 ( P < 0.01, P < 0.05) group, compared with control 1 and 2 group broilers. The MDA content in HS1 group was higher than that of in control 1 group broilers ( P < 0.05). Both the gene and protein expression levels of HSP70, TLR4 and NF-κB in the liver were significantly enhanced by heat stress. Furthermore, heat stress obviously enhanced the expression of IL-6, TNF-α, NF-κB P65, IκB and their phosphorylated proteins in the livers of broilers. In addition, heat stress promoted the activation of NLRP3 with increased NLRP3, caspase-1 and IL-1β levels. Conclusions These results suggested that heat stress can cause liver inflammation via activation of the TLR4-NF-κB and NLRP3 signaling pathways in broilers. With the extension of heat stress time, the effect of heat stress on the increase of NF-κB and NLRP3 signaling pathways tended to slow down.
 
Article
Background Pasteurella multocida is one of the most significant pathogens for a number of animals. In rabbits, the infection is generally associated with the P. multocida serogroups A and D, and the knowledge about the serogroup F is limited. In the present study, a P. multocida serogroup F isolate designated s4 was recovered from the lungs of rabbits died of respiratory disease in Fujian, in the southeast of China. The pathogenicity and genomic features of the s4 were then determined. Results The serotype and sequence type of s4 were F:L3 and ST12, respectively. The s4 was pathogenic for rabbits, but it was a low virulent strain comparing to the previously reported highly pathogenic P. multocida serogroup F strains J-4103, C21724H3km7, P-4218 and HN07. The whole genome of the s4 was then sequenced to understand the genomic basis for pathogenicity. Particularly, a large-sized fragment of approximate 275 kb in length was truncated from the chromosome to form a plasmid. Moreover, the in-frame deletion of natC and N-terminal redundance of gatF would resulted in the production of a mutant L3 outer core structure that was distinct from those of the other P. multocida strains belonging to the lipopolysaccharide genotype L3. We deduced that these features detected in the genome of s4 might impair the pathogenicity of the bacterium. Conclusions This study evaluated the pathogenicity and determined the genomic features of the rabbit sourced P. multocida serogroup F isolate s4, the observations and findings would helpful for the understanding of the pathogenicity variability and genetic diversity of P. multocida .
 
The result of the BAER examination of a cat with normal hearing in both ears
The result of the BAER examination of a cat with unilateral deafness
The result of the BAEP examination of a cat with bilateral deafness
Article
Background Data on sensorineural deafness (CSD) in purebred white client-owned cats is limited as most of the information on this disease entity is assured from mixed-breed experimental colonies. It is known that cats with blue irises are more predisposed to CSD having been described as a condition in which many structures in the inner ear are damaged resulting in hearing loss. Cats with CSD are born deaf or lose their hearing irreversibly within the first 4-5 weeks of life. It is important to diagnose cats with this hereditary condition in order to eliminate affected individuals from breeding. The objectives of this study were to ensure data on prevalence of CSD in a population of 72 client-owned purebred white cats in Poland according to the color of the irises and to determine if there are any predispositions with regard to CSD among different breeds of cats in which the dominant W gene is present. Results Conducted study included 72 purebred white cats from six different breeds. The prevalence of CSD in the conducted study was 16.7%, CI95 [8.9%; 23.3%]. Unilateral deafness (11.1%, CI95 [4.9%; 20.7%]) was more common than bilateral CSD (5.6%, CI95 [1.5%; 13.6%]). The studies did not show any association between sex and CSD, p = .46. No association between the blue color of irises and deafness in the studied population could be found, p = .91. When compared to the rest of the examined population, no association was found between CSD and a particular breed. Conclusions Overall prevalence of CSD regarding the examined population of purebred client-owned cats was reported as lower when compared to previous studies concerning purebred cats. Cats with blue irises are more likely to be deaf in accordance to the current state of knowledge, however in the conducted study, no significant association between the presence of blue irises and deafness in white purebred cats could be identified. In order to eliminate CSD from the population, it is necessary to conduct examinations and diagnose CSD in white cats with blue irises as well as with irises of color other than blue. Association between particular breed and CSD wasn’t identified.
 
Geographic location of Soure municipality (dark grey) within Marajó Island
Article
Background Marajó Island, within in the Amazon River Delta, supports numerous bands of feral equids including the genetically distinct Marajoara horses. Approximately 40% of the equids on the island are infected with Equine infectious anemia virus (EIAV). This high seropositivity rate coupled with the need to preserve rare breeds such as the Marajoara horse precludes euthanasia as the primary means for controlling EIAV in this region. In the absence of iatrogenic transmission, spread of this lentivirus is mediated primarily by hematophagous insects, whose year-round prevalence on the island is supported by favorable climatic conditions. In addition, cases of vertical EIAV transmission have been observed suggesting inclusion of seropositive mares in restorative breeding programs could result in their progeny becoming infected with this virus either pre-parturition or post-partum via hematophagous insects. Therefore, the aim of this study was to evaluate EIAV vertical and post-partum insect-mediated transmission rates among foals born to seropositive feral mares until natural weaning. Serum samples from foals born to seropositive feral mares within the Soure municipality, of Marajó Island, were collected to investigate their serological status, using an indirect ELISApgp45, with positive samples confirmed using the classical agar gel immunodiffusion (AGID) assay. Results The serological status of 28 foals were monitored over a 2-year period with some subjects, depending on their date of birth, being sampled up to six times. All foals remained with their respective mares until fully weaned at approximately 10 months of age. Only 2 foals (7.14%) in the study group became seropositive against EIAV. Conclusion The results demonstrate that in most cases it is possible to obtain seronegative foals born to and eventually weaned by EIA positive mares, even in equatorial regions where substantial rainfall and high temperatures favor the proliferation of insect vectors.
 
Choropleth maps indicating the proportion of dogs vaccinated for leptospirosis in 2016 in each region. Maps are at a NUTS level 1 region resolution. Map a) indicates the proportion of dogs in each region that are vaccinated against leptospirosis (with a leptospirosis vaccine of any level of serovar coverage), b) indicates the proportion of dogs in the regions that received a tetravalent leptospirosis vaccine and c) indicates the proportion of dogs in the regions that received a bivalent leptospirosis vaccine. Areas in darker green indicate higher proportion of vaccinated dogs
Results of mixed effects model examining significant variables associated with leptospirosis vaccination administration for dogs under primary veterinary care in 2016. The final model included neutering status, insurance status, top 20 breeds, owner IMD rank, biological age and corporate group. Interaction terms between age and neutering and age and insurance are shown. Missing data was retained and is labelled ‘missing’ here
Article
Background Leptospirosis is a zoonotic disease that is found globally and affects most mammalian species. Vaccination of dogs against leptospirosis is an important approach to preventing clinical disease, or reducing disease severity, as well as reducing transmission of the infection to humans. Although it is generally considered to be a ‘core’ vaccine, there is limited information on the level of leptospirosis vaccine usage and factors associated with its usage in dogs in the UK. The study aimed to report the uptake of leptospirosis vaccination and factors associated with its usage in a cohort of dogs under primary veterinary care during a 12-month period. Results From a population of 905,543 dogs, 49% (95%CI 48.9–49.1%) had at least one leptospirosis vaccine administered during the 12 months of study. Adult dogs had reduced odds of receiving a leptospirosis vaccine when compared to dogs < 1 year old, with dogs > 8 years old having a greater than ten-fold reduction in odds (OR = 0.08, 95%CI 0.07–0.09). Odds of receiving a leptospirosis vaccine was increased in insured dogs when compared to uninsured dogs (OR = 1.22, 95%CI = 1.17–1.28). Neutered dogs had reduced odds of receiving a leptospirosis vaccine (OR = 0.87, 95%CI 0.83–0.91). Breed associations with receiving a leptospirosis vaccine varied. Several breeds were associated with increased odds of receiving a leptospirosis vaccine when compared to crossbreed dogs, including Border Terriers (OR = 1.49, 95%CI 1.42–1.57), Golden Retrievers (OR = 1.30, 95%CI = 1.24–1.37), Cocker Spaniels (OR = 1.27, 95%CI 1.23–1.31) and West Highland White Terriers (OR = 1.27, 95%CI 1.22–1.31). French Bulldogs (OR = 0.64, 95%CI = 0.62–0.67), Staffordshire Bull Terriers (OR = 0.79, 95%CI 0.78–0.82) and Pugs (OR = 0.91, 95%CI =0.88–0.95) had significantly reduced odds of receiving a leptospirosis vaccination during the study. Conclusion This work identified that almost half of the UK primary care attending population received a leptospirosis vaccine during the year. Several demographic variables were associated with leptospirosis vaccine administration, with age being particularly important. Both the proportion of uptake and factors associated with leptospirosis vaccine usage can be used as a benchmark for comparisons in the future. Additionally, an understanding of which populations have reduced odds of receiving a leptospirosis vaccine can potentially be used for initiatives to encourage owner vaccination uptake in these groups.
 
Article
Background Adipose tissue (AT) is one of the most important mesenchymal stem cell (MSC) sources because of its high quantities, availability and ease of collection. After being collected samples, they should be transported to a laboratory for stem cell (SC) isolation, culture and expansion for future clinical application. Usually, laboratories are distant from animal husbandry centers; therefore, it is necessary to provide suitable conditions for adipose tissue transportation, such that adipose-derived MSCs are minimally affected. In the current study, the impact of tissue maintenance under different conditions on MSCs derived from these tissues was evaluated. We aimed at finding suitable and practical transportation methods in which ASCs go through the slightest changes. Results In the current study, after being collected, equine AT was randomized into eight groups: four samples were maintained in stem cell culture media at 25 οC and 4 οC for 6 and 12 hrs. as transportation via SC media groups. Three samples were frozen at three different temperatures (− 20, − 75 and − 196 οC) as cryopreserved groups; these samples were defrosted 1 week after cryopreservation. Fresh and unfrozen AT was evaluated as a control group. The tissue samples were then initiated into enzymatic digestion, isolation and the culturing of SCs. Cells at passage three were used to evaluate the ability to form colonies, proliferation rate, plotting of the cell growth curve, and viability rate. All experiments were performed in triplicate. Stem cell isolation was successful in all groups, although purification of SCs from the first series of cryopreservation at − 196 οC and two series of − 20 οC was unsuccessful. There was no significant difference between the surface area of colonies in all groups except for − 20 οC. The growth rate of transportation via stem cell media at 25 οC for 6 hrs. was similar to that of the control group. MTT analysis revealed a significant difference between 25 οC 12 hrs. Group and other experimental groups except for control, 4 οC 12 hrs. and − 196 οC group. Conclusion Data have shown freezing at − 75 οC, transportation via stem cell media at 4 οC for 12 hrs. and 25 οC for 6 hrs. are acceptable tissue preservation and transportation methods due to minor effects on MSCs features.
 
The map of Iran and sample collection sites
Phylogenetic tree of the ITS 2 fragments of rRNA gene for D. dendriticum isolated from livestock animals together with reference sequences. The phylogenetic tree represents that all sequences were grouped together and with the reference sequences. The phylogenetic tree was drawn using the maximum-likelihood method and the Tamura 3-parameter model. Bootstrap supports values of > 75% are indicated above the branches. Our sequences are indicated with maroon circle
Network analysis shows clusters of haplotypes for each D. dendriticum sequences based on A hosts and B haplotypes. Most of sequences are grouped as Hap 1. Each color indicates certain hosts. Plagiorchis elegans, which is indicated with black circle, is outgroup
Article
Background Dicrocoelium dendriticum is a broadly distributed zoonotic helminth, which is mainly reported from domesticated and wild ruminants. There is little data covering the molecular features of this trematode; therefore, current study aimed to molecularly analyze D. dendriticum in livestock. Methods Totally, 23 samples of D. dendriticum were collected from cattle, sheep, and goat from Ilam, Lorestan, and Khuzestan, three west and south-west provinces of Iran from February to August 2018. After genomic DNA extraction, the internal transcribed spacer (ITS) 2 fragment was amplified and sequenced in samples. To investigate genetic variations through the ITS 2 fragment of obtained D. dendriticum, phylogenetic tree and network analysis were employed. Results All 23 samples were successfully amplified and sequenced. Phylogenetic tree showed that our samples were clearly grouped in a clade together with reference sequences. There was no grouping based on either geographical regions or hosts. Network analysis confirmed the phylogenetic findings and showed the presence of nine distinct haplotypes, while our samples together most of sequences, which were previously submitted to the GenBank, were grouped in the Hap1. Conclusions Our findings indicated that although ITS 2 fragment discriminate D . dendriticum , this fragment is not suitable to study intra-species genetic variations. Therefore, exploring and describing new genetic markers could be more appropriate to provide new data about the genetic distribution of this trematode.
 
Assessment for eligibility
Article
Background Echocardiographic measurements may be influenced by breed-specific characteristics. Therefore, this study aims to establish reference values for standard echocardiographic measurements in pugs by investigating the influence of age, sex, heart rate, body weight and Brachycephalic Obstructive Airway Syndrome (BOAS). Sixty-two privately owned pugs underwent physical examination, blood sample collection, non-invasive blood pressure measurements and echocardiography. Influences of independent variables on echocardiographic measurements were examined using a multiple linear regression analysis model. For the entire study population, 95% prediction intervals were generated. Further, reference ranges for subcategories of clinical severities of BOAS were provided. Selected echocardiographic measurements of pugs were compared to reference values of previous studies generated from various breeds. Results In the study, a total of fifty-one privately owned pugs aged between two and 10 years were included for establishing reference ranges. Mainly body weight, but also age, sex and heart rate had influence on several echocardiographic parameters. The clinical grading of BOAS was conducted in 42 pugs. Except for pulmonic peak velocity (Pvel), which declined with increasing severity of BOAS, clinical symptoms of upper airway disease did not have significant impact on echocardiographic measurement results. Significant deviations, however, of left ventricular (LV) internal dimension (LVID), interventricular septum (IVS), LV posterior wall (LVPW), and tricuspid annular plane systolic motion excursion (TAPSE) compared to interbreed reference values were observed. Conclusions Breed-specific reference ranges for echocardiographic values with special regard to BOAS are provided to enable a more accurate assessment of cardiac health in pugs.
 
Article
Background Flagellin elicits potent immune response and may serve as a vaccine adjuvant. We previously reported that the N-terminus of flagellin (residues 1–99, n FliC) is sufficient for vaccine efficacy enhancement against Pasteurella multocida challenge in chickens. In this study, we futher tested the adjuvancy of n FliC in a subunit vaccine against the pig pathogen Actinobacillus pleuropneumoniae in a mice model. For vaccine formulation, the antigen ApxIIPF (the pore-forming region of the exotoxin ApxII) was combined with n FliC, either through genetic fusion or simple admixture. Results Immune analysis showed that n FliC, introduced through genetic fusion or admixture, enhanced both humoral (antibody levels) and cellular (T cell response and cytokine production) immunity. In a challenge test, n FliC increased vaccine protective efficacy to 60–80%, vs. 20% for the antigen-only group. Further analysis showed that, even without a supplemental adjuvant such as mineral salt or oil emulsion, genetically linked n FliC still provided significant immune enhancement. Conclusions We conclude that n FliC is a versatile and potent adjuvant for vaccine formulation.
 
Symptoms and Necropsy of Mycobacterium avium subsp. paratuberculosis infected Sheep. A Sheep with signs of diarrhea and emaciation. B Thickening and corrugation of the intestinal mucosa. C Enlarged and edematous abomasum. D Mesenteric lymph node edema and liquefaction. E Intestinal mucosa exfoliation. F Mesenteric congestion
PCR products from the IS900[L/AV] nested PCR. M: DNA marker 2000 bp. Lane 1-Lane3: Positive sample of feces; Lane 4-Lane3: Positive sample of mesenteric lymph node samples; Lane 7: negative sample
Pathological changes of the diseased sheep. A Myocardial fiber swelling, thickening and calcification. B Renal tubulointerstitial injuries . C Massive necrosis of spleen lymphocytes. D Thickening of the intestinal mucosa. E Necrosis of mesenteric lymph nodes. F Pathological changes of lung tissues
The total number (n) of animals at two different age groups (6 months and ≥ 1 year old) with the number of positive MAP samples, and the positive MAP detection rate (%) sampled from each herd in Bayannaoer, Inner Mongolia, China
Article
Background Paratuberculosis is a widespread chronic infection of Mycobacterium avium subspecies paratuberculosis (MAP) that causes significant economic losses to the sheep industry. The current study investigated this disease, which causes diarrhea in sheep, particularly, in Bayannaoer, Inner Mongolia, China. Diagnosis was based on clinical symptoms, pathological autopsy, histopathological inspection, and serological and molecular methods. Results MAP was confirmed using polymerase chain reaction using DNA extracted from tissue and fecal samples. Serum samples from 472 individual sheep were obtained to detect antibodies against MAP using an enzyme-linked immunosorbent assay. MAP antibodies were separately detected in 17.86% (35/196) and 18.48% (51/276) of sheep herds at approximately 6 months and ≥ 1 year of age, respectively. The tissue lesion and pathological section results were consistent with paratuberculosis infection. Conclusions To our knowledge, this is the first report of Mycobacterium avium subspecies paratuberculosis seroprevalence in Bayannaoer sheep in Inner Mongolia. Our findings show that MAP is not only prevalent, but also a potential threat to this region. Further investigations, including long-term epidemiological surveillance and isolation are needed for the awareness and effective treatment of paratuberculosis in sheep of Inner Mongolia.
 
Flow diagram of the selection and screening process
Changes in the volume of literature measuring appendicular skeletal muscle mass over time in each modality. DEXA, Dual-energy x-ray absorptiometry; MRI, Magnetic resonance imaging; CT, Computed tomography; US, Ultrasound; LC, Limb circumference
Recommendation of landmarks to determine the level of limb circumference measurement
Article
Background Monitoring changes in appendicular skeletal muscle mass is frequently used as a surrogate marker for limb function. The primary objective of this study was to review scientific information related to the assessment of appendicular skeletal muscle mass in dogs. The secondary objective was to develop practical recommendations for serial evaluation of muscle mass. Methods A scoping review was conducted with a systematic search of PubMed, Web of Science, CAB abstract, and Cochrane from inception to June 2021. The following modalities were included in the search: limb circumference, diagnostic ultrasound, computed tomography, magnetic resonance imaging, and dual-energy x-ray absorptiometry. Results A total of 62 articles that measured appendicular skeletal muscle mass in dogs were identified. Limb circumference (55 articles) was the most commonly used modality. Its reliability was investigated in five studies. Several factors, including measuring tape type, body position, joint angles, and the presence of hair coat, were reported as variables that can affect measurements. Diagnostic ultrasound (five articles) was validated in three articles, but there is scarce information about observer reliability and variables affecting the measurement. Computed tomography (four articles) and magnetic resonance imaging (one article) have been used to validate other modalities at a single time point rather than as a clinical tool for serial muscle mass monitoring. Dual-energy x-ray absorptiometry (two articles) has been used to quantify specific skeletal muscle mass but was mainly used to evaluate body composition in dogs. Conclusion Limb circumference and ultrasound are likely the main modalities that will continue to be used for serial muscle mass measurement in the clinical setting unless a new technology is developed. The reliability of limb circumference is questionable. Several key factors, including measuring tape type, body position, joint angles, and coat clipping, need to be controlled to improve the reliability of limb circumference measurements. Ultrasound may provide a reasonable alternative, but further studies are required to evaluate the reliability of this modality and identify factors that influence ultrasound measurements.
 
Article
Background Bisphenol-A (BPA) has estrogenic activity and adversely affects humans and animals' reproductive systems and functions. There has been a disagreement with the safety of BPA exposure at Tolerable daily intake (TDI) (0.05 mg/kg/d) value and non-observed adverse effect level (5 mg/kg/d). The current study investigated the effects of BPA exposure at various doses starting from Tolerable daily intake (0.05 mg/kg/d) to the lowest observed adverse effect level (50 mg/kg/d) on the testis development in male mice offspring. The BPA exposure lasted for 63 days from pregnancy day 0 of the dams to post-natal day (PND) 45 of the offspring. Results The results showed that BPA exposure significantly increased testis (BPA ≥ 20 mg/kg/d) and serum (BPA ≥ 10 mg/kg/d) BPA contents of PND 45 mice. The spermatogenic cells became loose, and the lumen of seminiferous tubules enlarged when BPA exposure at 0.05 mg/kg/d TDI. BPA exposure at a low dose (0.05 mg/kg/d) significantly reduced the expression of Scp3 proteins and elevated sperm abnormality. The significant decrease in Scp3 suggested that BPA inhibits the transformation of spermatogonia into spermatozoa in the testis. The RNA-seq proved that the spliceosome was significantly inhibited in the testes of mice exposed to BPA. According to the RT-qPCR, BPA exposure significantly reduced the expression of Snrpc (BPA ≥ 20 mg/kg/d) and Hnrnpu (BPA ≥ 0.5 mg/kg/d). Conclusions This study indicated that long-term BPA exposure at Tolerable daily intake (0.05 mg/kg/d) is not safe because low-dose long-term exposure to BPA inhibits spermatogonial meiosis in mice testis impairs reproductive function in male offspring.
 
Prevalence and distribution of coccidiosis in Korean chicken farms. A Frequency of the number of oocysts per gram of fecal samples. B Specific rate and number of Eimeria species present in positive sample. C Distribution of Eimeria species in samples
Status of coccidiosis based on level of infection and Eimeria species. A Level of infection based on the number of oocysts per gram of fecal samples. B Mean of numbers of Eimeria species present in positive samples. C Species-specific distribution of Eimeria species based on the source of fecal samples
Comparison of body weight gain in field sample-infected chickens. Five-day-old ROSS 308 female chickens were orally infected with 3 × 10⁴ sporulated oocysts of 9 different field samples (A–I). Body weight gain (n = 20) was measured at 9 days after infection. * P < 0.05, ** P < 0.01, and ***P < 0.001 indicate significant differences compared to untreated and healthy group (NC). The results represent mean ± SE values. NC, negative control; PC, untreated and infected group as a positive control; A-I, farm samples
Comparison of oocyst numbers in field sample-infected chickens. Five-day-old ROSS 308 female chickens were orally infected with 3 × 10⁴ sporulated oocysts of 9 different field samples (A–I). Production of oocysts per group (n = 20) was obtained from the fecal samples collected from days 6 to 9 post-infection. * P < 0.05, ** P < 0.01, and ***P < 0.001 indicate significant differences compared to untreated and infected group (PC). The results represent mean ± SE values. NC, negative control; PC, untreated and infected group as a positive control; A-I, farm samples
Comparison of intestinal lesion scores in field sample-infected chickens. Five-day-old ROSS 308 female chickens were orally infected with 3 × 10⁴ sporulated oocysts of 9 different field samples (A–I). At day 7 post-infection, five chickens from each group were randomly selected for intestinal lesion scoring. Lesions scores range from 0–4, following the Johnson and Reid [24] scoring technique. * P < 0.05 indicates significant difference compared to untreated and infected group (PC). Results represent mean ± SE values. NC, negative control; PC, untreated and infected group as a positive control; A-I, farm samples
Article
Background Coccidiosis is a poultry disease that occurs worldwide and is caused by Eimeria species. The infection is associated with reduced feed efficiency, body weight gain, and egg production. This study aimed to investigate the current status of coccidiosis and anticoccidial resistance to anticoccidial drugs used as part of control strategies for this disease in Korean chicken farms. Results An overall prevalence of 75% (291/388) was found. Positive farms contained several Eimeria species (mean = 4.2). Of the positive samples, E. acervulina (98.6%) , E. maxima (84.8%) , and E. tenella (82.8%) were the most prevalent species. Compared with cage-fed chickens, broilers and native chickens reared in free-range management were more at risk of acquiring an Eimeria infection. Sensitivities to six anticoccidial drugs (clopidol, diclazuril, maduramycin, monensin, salinomycin, and toltrazuril) were tested using nine field samples. Compared with untreated healthy control chickens, the body weight gains of infected chickens and treated/infected chickens were significantly reduced in all groups. Fecal oocyst shedding was significantly reduced in four clopidol-treated/infected groups, three diclazuril-treated/infected groups, two toltrazuril-treated/infected groups, one monensin-treated/infected group, and one salinomycin-treated/infected group, compared with the respective untreated/infected control groups. Intestinal lesion scores were also reduced in three clopidol-treated/infected groups, one monensin-treated/infected group, and one toltrazuril-treated/infected group. However, an overall assessment using the anticoccidial index, percent optimum anticoccidial activity, relative oocyst production, and reduced lesion score index found that all field samples had strong resistance to all tested anticoccidial drugs. Conclusion The results of this large-scale epidemiological investigation and anticoccidial sensitivity testing showed a high prevalence of coccidiosis and the presence of severe drug resistant Eimeria species in the field. These findings will be useful for optimizing the control of coccidiosis in the poultry industry.
 
The duodenum motility in healthy donkeys (Equus asinus) after IV administration of isotonic saline or dexmedetomidine at 3, 5, and 7 μg/kg. Each point represents the number of contractions (contraction / 3 minutes) expressed as mean ± standard deviation (SD) at different time points zero, 15, 30, 45, 60, 90, and 120 minutes post-administration. *: Mean ± SD with a superscript asterisk at the same time point are significantly different at P < 0.01
The jejunal motility in healthy donkeys (Equus asinus) after IV administration of isotonic saline or dexmedetomidine at 3, 5, and 7 μg/kg. Each point represents the number of contractions (contraction / 3 minutes) expressed as mean ± standard deviation (SD) at different time points zero, 15, 30, 45, 60, 90, and 120 minutes post-administration. *: Mean ± SD with a superscript asterisk at the same time point are significantly different at P < 0.01
The left colonic motility in healthy donkeys (Equus asinus) after IV administration of isotonic saline or dexmedetomidine at 3, 5, and 7 μg/kg. Each point represents the number of contractions (contraction / 3 minutes) expressed as mean ± standard deviation (SD) at different time points zero, 15, 30, 45, 60, 90, and 120 minutes post-administration. *: Mean ± SD with a superscript asterisk at the same time point are significantly different at P < 0.01
The right colonic motility in healthy donkeys (Equus asinus) after IV administration of isotonic saline or dexmedetomidine at 3, 5, and 7 μg/kg. Each point represents the number of contractions (contraction / 3 minutes) expressed as mean ± standard deviation (SD) at t different time points zero, 15, 30, 45, 60, 90, and 120-minutes post-administration. *: Mean ± SD with a superscript asterisk at the same time point are significantly different at P < 0.01
The cecum motility in healthy donkeys (Equus asinus) IV administration of isotonic saline or dexmedetomidine at 3, 5, and 7 μg/kg. Each point represents the number of contractions (contraction / 3 minutes) expressed as mean ± standard deviation (SD) at t different time points zero, 15, 30, 45, 60, 90, and 120 minutes post-administration. *: Mean ± SD with a superscript asterisk at the same time point are significantly different at P < 0.01
Article
Aim Gastrointestinal effects of different doses of dexmedetomidine in donkeys are still unidentified. The current study aimed to evaluate the impact of different doses of dexmedetomidine on the motility of selected parts of the gastrointestinal tracts in donkeys using transabdominal ultrasonography. Materials and methods An experimental crossover study was conducted on 30 healthy donkeys of both sexes (15 males and 15 females; 160 ± 60 kg). With a two-week washout period, each donkey received an injection of either a normal saline solution or three different doses of dexmedetomidine (3, 5, and 7 μg/kg, respectively). All medications were administered intravenously in equal volumes. The contractility of selected intestinal segments (duodenum, jejunum, left colon, right colon, and cecum) was measured 3 min before administration (zero time) and at 15, 30, 45, 60, 90, and 120 minutes after administration. Results Small and large intestinal motility was within the normal ranges before IV injection of normal isotonic saline or dexmedetomidine at a dose of 3, 5, and 7 μg/kg. Two Way Repeated Measures ANOVA output of the data displayed a statistically significant the between time and treatments for the contractility of each of the duodenum ( P = 0.0029), jejunum ( P = 0.0033), left colon ( P = 0.0073), right colon ( P = 0.0035), and cecum ( P = 0.0026), implying that the impact of treatment on the gastric motility varied among different time points. The simple main effect analysis revealed that the IV dexmedetomidine at 3, 5, and 7 μg/kg doses significantly inhibited ( P ≤ 0.01) the bowel contractility compared to the administration of isotonic saline. Conclusion Dose-dependent inhibitory effect of dexmedetomidine on intestinal motility was reported in donkeys following intravenous administration. This inhibitory effect on intestinal motility should be considered in clinical practice .
 
Article
Background Feline injection-site sarcomas (FISSs) are malignant mesenchymal tumors of different histotypes. The pathogenesis of FISS has been correlated with chronic inflammation, resulting in neoplastic transformation. Activation of the Janus kinase-signal transducer and activator of transcription 3 (STAT3) have been demonstrated to play a critical role in tumor development by regulating signaling pathways involved in cell proliferation, survival, metastasis, and angiogenesis in human medicine. To characterize the role of STAT3 in FISS, we first detected STAT3 and phosphorylated STAT3 in formalin-fixed and paraffin-embedded (FFPE) FISS tissues using immunohistochemical staining. Results STAT3 was detected in 88.9% (40/45) of FISS cases, and phosphorylated STAT3 was detected in 53.3% (24/45) of cases. However, the expression levels of both forms of STAT3 were not correlated with tumor grade. To study the role of STAT3 in tumor survival, two primary cells derived from FISSs of two cats exhibiting consistent immunophenotypes with their parental FFPE tissues were established. A dose-dependent inhibitory effect on cell proliferation was observed in both primary FISS cells treated with the STAT3 inhibitor, 5-hydroxy-9,10-dioxo-9,10-dihydroanthracene-1-sulfonamide. Conclusions The STAT 3 may play an important role in the tumorigenesis of FISS and be a potential molecular therapeutic target for FISS.
 
(continued)
Color obtained after the centrifugation of the Salivette tubes once a cows’ saliva pool and deionized water were submitted to incubation during 5 min at 38 °C (C2) and with 250 mg of feed-based on a standard total mixed ration usually offered to lactation cows in production (F), wheat hay (H), and grass (G)
Coefficient of variations (CVs) of the means (n = 5) from the specimens C2 (cows’ saliva pool submitted to incubation during 5 min at 38 °C), F, H, and G (cows’ saliva pool submitted to incubation with 250 mg of feed-based a standard total mixed ration, wheat hay, and grass, respectively) compared to their control clean saliva pool (specimen C1)
In vitro experimental workflow performed for the food effect evaluation in a profile of salivary analytes measurement in pool saliva from five cows during five different days. *Standard feed based on a total mixed ration (broccoli -remains-, oat silage, corn -ground-, barley -bagasse-, alfalfa silage, wet corn pulp, rapeseed, dehydrated alfalfa, dry beet pulp, non-protein hydrogen source, barley straw, and a mineral pre-mix)
Article
Background The effect in a sialochemistry profile of the presence of usually available feed in dairy cows was evaluated by an in vitro experiment. For this purpose, a pooled clean saliva from five healthy dairy cows was incubated five times with a standard feed based on a total mixed ration (F), wheat hay (H), and grass (G). The salivary panel was integrated by biomarkers of stress (cortisol -sCor-, salivary alpha-amylase -sAA-, butyrylcholinesterase -BChE-, total esterase -TEA-, and lipase -Lip-), immunity (adenosine deaminase -ADA-), oxidative status (Trolox equivalent antioxidant capacity -TEAC-, the ferric reducing ability of saliva -FRAS-, the cupric reducing antioxidant capacity -CUPRAC-, uric acid, and advanced oxidation protein products -AOPP-), and enzymes, proteins, and minerals of general metabolism and markers of liver, muscle, and renal damage (aspartate aminotransferase -AST-, alanine aminotransferase -ALP-, γ-glutamyl transferase -gGT-, lactate dehydrogenase -LDH-, creatine kinase -CK-, creatinine, urea, triglycerides, glucose, lactate, total protein, phosphorus, and total calcium). Results Most of the evaluated analytes showed a coefficient of variations (CV) higher than 15% and/or significant changes compared with the clean saliva when feed was present. Some analytes, such as the oxidative status biomarkers (CV > 80%), AST (CV > 60%), or glucose (CV > 100%), showed significant changes with all the feed types tested. Others showed significant differences only with certain types of feed, such as LDH with F (CV > 60%) or triglycerides with F (CV > 100%) and H (CV > 95%). However, sCor or gGT remained unchanged (CV < 15%, P > 0.05) in all the treatments. Conclusions The presence of feed can produce changes in most of the analytes measured in cows’ saliva, being of high importance to consider this factor when saliva is used as a sample to avoid errors in the interpretation of the results.
 
A typical P-VEP waveform obtained under sedation with butorphanol and dexmedetomidine (A) and general anesthesia with propofol and sevoflurane (B), recorded from the Oz electrode
A typical F-VEP waveform obtained under sedation with butorphanol and dexmedetomidine (A) and general anesthesia with propofol and sevoflurane (B), recorded from the Oz electrode
Electrode placement for VEP recording. The recording electrodes were placed over the inion, or dorsal occipital protuberance (O1, Oz, O2). The reference electrode was placed at the level of the forehead (Fpz). The ground electrode (Cz) was placed halfway between Oz and Fpz
Pattern visual evoked potential (P-VEP) peak latencies
Pattern visual evoked potential (P-VEP) peak amplitudes
Article
Background Visual evoked potentials (VEPs) can provide objective functional assessment of the post-retinal visual pathway. This study compared the effects of sedation (butorphanol and dexmedetomidine) and general anesthesia (propofol and sevoflurane) on pattern and flash VEPs. Dogs ( n = 13) underwent sedation or anesthesia and VEPs were obtained from 3 subcutaneous recording electrodes placed on the head (O1, Oz, O2). Results Pattern VEPs could only be recorded under sedation and a maximum of 3 peaks were identified (N75, P100, N135). Flash VEPs could be recorded under both sedation and anesthesia and a maximum of 5 peaks were identified (N1, P1, N2, P2, N3). The latency of the N1 peak and the baseline-N1 amplitude were significantly longer under general anesthesia. Conclusion Visual evoked potentials should be preferentially recorded in dogs sedated with dexmedetomidine and butorphanol, regardless of the stimulus.
 
Dendrogram of PFGE-SmaI profiles of S. epidermidis isolates from untreated and treated cows with homeopathy. T = treated cows; U = untreated cows; STA = strong adherent; WEA = weak adherent; ST = sequence types
Dendrogram of the PFGE-SmaI profiles of isolates of S. chromogenes from untreated and treated cows with homeopathy. T = treated cows; U = untreated cows; STA = strong adherent; WEA = weak adherent; NA = nonadherent
Dendrogram of the PFGE-SmaI profiles of S. aureus isolates from untreated and treated cows with homeopathy. T = treated cows; U = untreated cows; STA = strong adherent; WEA = weak adherent; NA = nonadherent; ST = sequence types
Article
Background Mastitis is one of the major diseases in dairy cattle, as it causes great economic losses to producers due to the reduction of milk production and changes in the quality of the product. The disease is mainly caused by bacteria of the genus Staphylococcus spp., these microorganisms can express various virulence factors, such as biofilms for example. In herds with organic management, producers and technicians use unconventional ways to treat and control the disease, such as homeopathy. However, it is not known if this type of treatment is able to control pathogenic bacteria such as those of the genus Staphylococcus , of relevance to animal and human health. Thus, the objective of this study was to investigate the production of biofilm in vitro and its genes by Staphylococcus spp. isolated in the milk of cows treated with homeopathy, as well as the persistence of microorganisms in animals. Methods Ninety-nine isolates of Staphylococcus spp. from cows treated and not treated with homeopathy were identified by internal transcribed space-polymerase chain reaction and investigated for the presence of the ica ABCD, bap , aap , atlE , and bhp genes and in vitro biofilm production using the adhesion method on polystyrene plates. The enzyme restriction profile was determined by Pulsed-Field Gel Electrophoresis. Clusters of S. aureus and S. epidermidis with three or more isolates had an isolate selected for Multilocus Sequence Typing. Results The frequency of S. aureus isolations was similar in treated and untreated cows, while 71.4% of the coagulase-negative identified were isolated in cows treated with homeopathy. The distribution of the operon ica genes was similar in animals with and without treatment, except for the ica D gene, more frequent in treated cows. Production of biofilm was associated with presence of one or more genes from the ica ADBC operon. S. aureus revealed a greater diversity and greater dissemination in cows treated and not treated with homeopathy. Sequence Types ST1, ST5, and ST126 were identified in S. aureus . Conclusions The presence of biofilm-associated genes and the in vitro production of biofilms, combined with the persistence of clonal profiles of Staphylococcus spp. demonstrate other forms of control for bovine mastitis should be researched for organic production herds.
 
Article
Background As the frequency of spine surgery increases in the veterinary field, many studies have been conducted on minimally invasive spine surgery (MISS). Although many studies have been conducted on the thoracolumbar spine about MISS in animals, several problems and limitations have emerged regarding this method. Therefore, we developed a three-dimensional (3D) printed patient-specific surgical guide (3DPSSG) using 3D printing technology to overcome these problems. We aimed to evaluate the accuracy and safety of the 3DPSSG in minimally invasive mini-hemilaminectomy-corpectomy (MI-MHC). MI-MHC using 3DPSSG and an endoscopic system was performed at L1–L2 in 15 cadaveric dogs. The procedure of fixing the surgical guide to the vertebral body through screws and the surgical procedure using the guide were performed by two surgeons with different experiences. Postoperative computed tomography was used to measure planned and postoperative screw trajectories (angle, protruding from the far cortex) and to create 3D rendering images of vertebrae to evaluate the direction of bone window formation, corpectomy slot length, depth, and height ratio. Results The two groups which performed by two surgeons with different experiences did not differ in terms of screw angle deviation and length of the screw protruded from the far cortex. The corpectomy slot-length ratio was not different between the two groups; however, the slot-depth and height ratios were different. Conclusions No differences were detected in screw trajectory and corpectomy slot-length ratio between the two groups. The 3DPSSG for MI-MHC is classified as accurate and safe; therefore, it can be an alternative to the conventional technique in dogs.
 
Article
Background Salmonella is a leading foodborne and zoonotic pathogen, and is widely distributed in different nodes of the pork supply chain. In recent years, the increasing prevalence of antimicrobial resistant Salmonella poses a threat to global public health. The purpose of this study is to the prevalence of antimicrobial resistant Salmonella in pig slaughterhouses in Hubei Province in China, and explore the effect of using lytic bacteriophages fighting against antimicrobial resistant Salmonella . Results We collected a total of 1289 samples including anal swabs of pigs (862/1289), environmental swabs (204/1289), carcass surface swabs (36/1289) and environmental agar plates (187/1289) from eleven slaughterhouses in seven cities in Hubei Province and recovered 106 Salmonella isolates. Antimicrobial susceptibility testing revealed that these isolates showed a high rate of antimicrobial resistance; over 99.06% (105/106) of them were multidrug resistant. To combat these drug resistant Salmonella , we isolated 37 lytic phages using 106 isolates as indicator bacteria. One of them, designated ph 2–2, which belonged to the Myoviridae family, displayed good capacity to kill Salmonella under different adverse conditions (exposure to different temperatures, pHs, UV, and/or 75% ethanol) and had a wide lytic spectrum. Evaluation in mouse models showed that ph 2–2 was safe and saved 80% (administrated by gavage) and 100% (administrated through intraperitoneal injection) mice from infections caused by Salmonella Typhimurium . Conclusions The data presented herein demonstrated that Salmonella contamination remains a problem in some pig slaughter houses in China and Salmonella isolates recovered in slaughter houses displayed a high rate of antimicrobial resistance. In addition, broad-spectrum lytic bacteriophages may represent a good candidate for the development of anti-antimicrobial resistant Salmonella agents.
 
Relative expression values of NFKB1, NFKB2, RELA, RELB and REL mRNA in the inguinal lymph nodes from non-pregnant ewes and pregnant ewes (n = 6 for each group). The mean mRNA expression level for each target gene was normalized to the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and the mean ΔCt value for the tissues collected from the cyclic ewe was a calibrator to compare the expression values from different stages of gestation. Note: DN16 = day 16 of the estrous cycle; DP13 = day 13 of pregnancy; DP16 = day 16 of pregnancy; DP25 = day 25 of pregnancy. Significant differences (P < 0.05) are indicated by different letters
Expression of NF-κB p105, NF-κB p100, p65, RelB and c-Rel proteins in the inguinal lymph nodes from non-pregnant ewes and pregnant ewes (n = 6 for each group). The mean band intensity was normalized to the band intensity of GAPDH protein, and the relative protein expression level for each target protein from the cyclic ewe was a calibrator to compare the expression values from different stages of gestation. Note: DN16 = day 16 of the estrous cycle; DP13 = day 13 of pregnancy; DP16 = day 16 of pregnancy; DP25 = day 25 of pregnancy. Significant differences (P < 0.05) are indicated by different letters within the same color column
Representative immunohistochemical localization of p65 protein in the inguinal lymph nodes from non-pregnant ewes and pregnant ewes (n = 6 for each group). Lymph node is divided into an outer cortex (CO) and an inner medulla (ME). Lymph enters the convex through the subcapsular sinus (SS) and trabeculae (TR) around the lymphoid nodules (LN), and flows into the medulla through the lymph sinus (LS) around the medullary cord (MC). Note: HE = stained by haematoxylin and eosin; Clt = Negative control; DN16 = day 16 of the estrous cycle; DP13 = day 13 of pregnancy; DP16 = day 16 of pregnancy; DP25 = day 25 of pregnancy. Bar = 20 μm
Article
Background Pregnancy-induced immunological changes contribute to the maternal immune tolerance. Nuclear factor kappa B (NF-κB) pathway participates in regulating both innate and adaptive immunities, and lymph nodes play key roles in adaptive immune reaction. However, it is unclear whether early pregnancy changes the expression of NF-κB family in maternal lymph node in sheep. Methods In this study, the samples of inguinal lymph nodes were collected from ewes on day 16 of the estrous cycle, and on days 13, 16 and 25 of pregnancy, and expression of NF-κB family, including NF-κB p105 ( NFKB1 ), NF-κB p100 ( NFKB2 ), p65 ( RELA ), RelB ( RELB ) and c-Rel ( REL ), were analyzed through real-time quantitative PCR, Western blot and immunohistochemical analysis. Results The expression levels of NF-κB p105 and c-Rel downregulated, but NF-κB p100 upregulated on day 25 of pregnancy. The expression levels of p65, RelB and c-Rel peaked at day 13 of pregnancy, and expression level of RelB was higher during early pregnancy comparing to day 16 of the estrous cycle. In addition, p65 protein was located in the subcapsular sinus and lymph sinuses. Conclusion This paper reported for the first time that early pregnancy has effects on the expression of NF-κB family, which may contribute to the maternal immunoregulation through blood circulation and lymph circulation during early pregnancy in sheep.
 
Article
Background Mammary gland tumours are the most frequently diagnosed tumours in the female dogs but just a few studies have analysed their epidemiology. Therefore, we set out to describe the epidemiology of canine mammary cancer in the Canary Archipelago, Spain. We analysed a pathology tumour registry (PTR) and identified 7362 samples obtained from 5240 female dogs resident on the Canary Archipelago during an 18-year period (2003–2020). Using a case–control study design, we compared mammary tumour affected dogs with the Canarian canine population registry in order to elucidate the breed associations for these tumours. Results The frequency of a diagnosis of mammary tumours relative to all tumour diagnoses in female dogs decreased during the study period from 62.7% to 48.9%. Contemporaneously, the proportion of dogs diagnosed with mammary tumours who were also neutered increased from 13.6% to 26.9%. There was a negative correlation ( R = -0.84) between these changes. Additional findings were that: the proportion of female dogs diagnosed with multiple tumours increased by 23.5% and that the proportion of malignant tumours 89.2% diagnosed has remained stable through the period. Benign mammary tumours were diagnosed at younger ages (9.2 years old) than carcinomas (9.7 years old) and sarcomas (10.4 years old). Epithelial mammary tumours were diagnosed at younger ages in entire female dogs. Samoyed, Schnauzer, Poodle, German Pinscher and Cocker Spaniel were the breeds with the highest odds-ratios (OR) in comparison with the reference (crossbreeds) while Miniature Pinscher, American Staffordshire Terrier, English Pointer as well as some local breeds such as the Canary Warren Hound and the Majorero had the lowest ORs. Conclusions This study provides a description of the changing epidemiology of canine mammary cancer in the Canary Archipelago over the last two decades. We found high rates of CMT with a significant predominance of malignant tumours. Exact risk factors are uncertain, but a combination of environmental, regional socioeconomic affecting human and their pets, and animal management factors are likely to play a part. Specifically, neutering was negatively associated with the proportion of epithelial mammary gland tumours and breeds native to the region were at lower risk of mammary tumours. A deeper analysis of all these factors will facilitate a deeper understanding of the epidemiology of mammary gland tumours in both the canine and the human population.
 
Article
Background Autologous conditioned serum (ACS) has been extensively used in the field of veterinary orthopaedics and sports medicine. Due to the autologous and blood-derived nature of this product, issues such as individual variability, need for storage at low temperatures and non-availability for immediate are frequently encountered for ACS use in the field. To address those issues, we proposed the evaluation of an off-the-shelf allogeneic freeze-dried version of conditioned serum in an in vitro model of osteoarthritis. In this study, we evaluated if origin (autologous and allogeneic) and preparation (frozen and freeze-dried) of conditioned serum could influence in its effect in an in vitro model. Results IL-1β stimulation in cartilage led to a significant increase in media GAG and decreased levels of GAG in cartilage explants at the termination of the experiment. No significant differences were noted in outcomes measured in the cartilage explants with respect to the main effects of treatment (frozen versus freeze-dried serum), autologous versus allogeneic preparations or based on serum concentration. Conclusions The study did not observe any substantial differences in the response of cartilage to allogeneic freeze-dried CS when compared to other independent parameters (autologous and frozen preparations). Further investigation using in vivo systems appears warranted.
 
Initial physical examination. A 25-year-old donkey in poor body condition with severe generalised alopecic and exfoliative dermatitis (a); close up of the skin, which was generally thin, covered by large quantities of large, thin, greyish scales (b), occasionally eroded with sero-haemorrhagic exudate (c). The hoofs were not affected (d). The lips and muzzle were very scaly (e)
Cytological examination. A direct skin smear was taken from eroded lesions on the lateral thorax. Degenerate neutrophils, phagocytized cocci and keratinocytes (green cross) were observed. (Stained with RAL®555, RAL Diagnostics; Site Montesquieu-Martillac, France. Magnification × 1000, bar = 10 µm)
Histopathological examination of biopsies taken from the lateral thorax. Interface dermatitis (a) with massive exocytosis in the epidermis of a homogenous population of lymphoid cells showing atypia (b). Clusters of neoplastic cells were present within the epidermis forming Pautrier microabscesses (c); [H&E staining, magnification × 40 (a), × 200 (b) and × 400 (c), bars = 1000 µm (a), 100 µm (b) and 10 µm (c)]. Immunohistochemical staining for CD3 showed uniform labelling of the neoplastic cells for CD3 (d), whereas staining for CD20 was negative (e). The Ki-67 labelling fraction was hard to quantify in the epidermis as the background labelling of basal epithelial cells was prominent, but by comparing the labelling fraction in the superficial dermis, it was estimated at less than 20% (f). (Magnification × 400, bar = 10 µm)
Electron microscopy of a skin biopsy taken from the lateral thorax showing multiple lymphocytes with convoluted (a) or cerebriform (b) nuclei. Prominent nucleoli were observed in the nucleus (n). Scale bars = 1 µm
Capillary electrophoresis traces of PCR for antigen receptor rearrangement (PARR). T-cell receptor gamma genes (TRG) rearrangement analysis of blood from a normal 8-month old donkey (a), blood (b) and skin (c) of the donkey with mycosis fungoides. a and b: TRG polyclonal; c:. TRG clonal in a polyclonal background—clonal peaks are at 76 bp and 80 bp. The left most peak in all trace files (a, b and c) is the 50 bp calibration marker
Article
Background Cutaneous epitheliotropic T-cell lymphoma is a malignant tumour of the skin already reported in humans, dogs, cats, horses, and other species, but not previously in donkeys. The standard diagnosis is based on clinical, morphological and immunophenotypic data. Differentiation of malignant versus benign proliferation of lymphocytes is crucial; in ambiguous cases T-cell receptor gamma (TRG) molecular clonality should be tested. In the present paper, we report a case of mycosis fungoides diagnosed in a donkey whose diagnosis was based on clinical, histological and immunohistochemical aspects and a positive TRG clonality test. Case presentation A twenty-five-year-old donkey gelding was referred with a mildly pruritic, generalised and severe exfoliative dermatosis. Otherwise, the animal was clinically healthy, though mildly underweight. Dermatological examination revealed severe generalised alopecic and exfoliative dermatitis, occasionally eroded, with high number of large, thin, greyish scales. All mucocutaneous junctions except the hoofs were affected. Ectoparasites and dermatophytes were ruled out. The complete blood count and blood smear evaluation revealed mild normocytic normochromic anemia. The biochemistry panel showed mild hyperproteinemia with albumin within the normal range. Protein electrophoresis showed moderate polyclonal hypergammaglobulinemia. Histological findings were characterised by interface dermatitis with massive exocytosis in the epidermis of a homogenous population of lymphoid cells showing atypia. Clusters of neoplastic cells were present within the epidermis forming Pautrier “microabscesses”. These findings are consistent with cutaneous epitheliotropic lymphoma. Immunohistochemical staining revealed uniform labelling of the neoplastic cells for CD3, and lack of expression of CD20 (a B cell lineage associated marker). Molecular clonality PCR (PARR) was performed using equine TRG primers; this revealed a clonal rearrangement in a heavy polyclonal background. Transmission electronic microscopy showed multiple lymphocytes with convoluted or cerebriform nuclei. Conclusions This case report provides the first evidence of clinical, histopathological, immunophenotypic features, electron microscopy findings and molecular analysis of a cutaneous epitheliotropic T-cell lymphoma (mycosis fungoides) in a donkey. Our observations suggest that cutaneous T-cell lymphoma should be included in the differential diagnoses of exfoliative dermatitis, even those progressing in a chronic pattern and/or with few or no pruritus.
 
Mean ± standard deviation heart rates (HR) in horses during isoflurane general anaesthesia with dexmedetomidine administered as a constant rate infusion (group CRI) or by repeated subcutaneous administration (group SC)
Mean ± standard deviation systolic arterial blood pressures (SAP) in horses during isoflurane general anaesthesia with dexmedetomidine administered as a constant rate infusion (group CRI) or by repeated subcutaneous administration (group SC)
Mean ± standard deviation mean arterial blood pressures (MAP) in horses during isoflurane general anaesthesia with dexmedetomidine administered as a constant rate infusion (group CRI) or by repeated subcutaneous administration (group SC). Significant time points between groups (p < 0.05) are indicated with an asterisk (*)
Mean ± standard deviation diastolic arterial blood pressures (DAP) in horses during isoflurane general anaesthesia with dexmedetomidine administered as a constant rate infusion (group CRI) or by repeated subcutaneous administration (group SC). Significant time points between groups (p < 0.05) are indicated with an asterisk (*)
Article
Background A balanced anaesthetic protocol is a common concept in modern veterinary anaesthesia and aims to maintain good intraoperative cardiopulmonary function. In horses, alpha-2-agonists produce sedation and analgesia and have been shown to reduce inhalational anaesthetic requirements when administered intravenously. Furthermore, these drugs can improve recovery quality. Preliminary investigations of subcutaneous dexmedetomidine administration in humans demonstrated a reduced haemodynamic impact if compared with the intravenous route suggesting that dexmedetomidine is adequately absorbed with both administration routes. The aim of the study was to compare two different dexmedetomidine (DEX) administration routes: intravenous constant rate infusion (CRI) versus repeated subcutaneous (SC) injections on cardiopulmonary function and recovery in anaesthetized horses. Results No significant differences between groups in heart rate and systolic arterial pressure were detected. A significantly higher mean and diastolic arterial pressure were detected in the SC group at T25 (p = 0.04; p = 0.02), T75 (p = 0.02; p = 0.009), and T85 (p = 0.001; p = 0.005). In SC group there was a significantly lower dobutamine infusion rate (p = 0.03) and a significantly higher urinary output (p = 0.02). Moreover, recovery quality was higher (p = 0.01). Conclusions Cardiopulmonary effects in both groups were comparable and within clinical ranges with less dobutamine requirement in the subcutaneous group. Recovery was of better quality with fewer attempts in horses receiving subcutaneous dexmedetomidine. The present study suggests that intravenous constant rate infusion and subcutaneous repeated administration of dexmedetomidine at indicated dosage can be useful in balanced anaesthesia without any systemic or local adverse effects; moreover, in healthy horses undergoing general anaesthesia, repeated subcutaneous dexmedetomidine administration may be a suitable alternative if constant rate infusion is not feasible.
 
Map of Sardinia showing the collection of S. uberis isolates from sheep mastitis milk samples. Each point represents an individual isolate while point colour represents the PopPUNK cluster number as an alternative for MLST typing. PopPUNK cluster also shown in white above point. Regions are shown in bolded black text
Mid-rooted neighbour-joining tree output from PopPUNK showing sheep mastitis isolates (this study) in the context of S. uberis from other sources. Nodes are coloured based on their assigned PopPUNK cluster, and shaped based on host. Heatmap indicates broad geographic location. Grey overlay box shows monophyletic Sardinian sheep clade
Heatmap showing antibiotic resistance profiles of all 46 S. uberis isolates, sorted by PopPUNK cluster. Isolates are defined as either ‘Sensitive’, ‘Intermediate’ or ‘Resistant’, reflected by increase of colour saturation. The PopPUNK clusters are annotated by the rainbow colour palette. Local location of isolate is also shown. Figure adapted from (15)
Gene cluster comparison of the phage recombinase-mediated multi-antimicrobial resistance cassette. Phage proteins are coloured in blue, with dark blue indicating the recombinase. Pink and red colours indicate antimicrobial resistance genes. Grey genes show cluster flanking genes. Black shows genes of unknown function
Heatmap of putative virulence factors comparing S. uberis isolated from this study to Australian isolates. Virulence factor broad ‘type’ is shown based on colour, and transparency of colour denotes presence or absence within a genome
Article
Background Streptococcus uberis is one of the main causative agents of ovine mastitis, however little is known about this global, environmental pathogen and its genomic mechanisms of disease. In this study, we performed genomic analysis on 46 S. uberis isolates collected from mastitis-infected sheep in Sardinia (Italy). Results Genomes were assigned into lineage clusters using PopPUNK, which found 27 distinct isolate clusters, indicating considerable genetic variability consistent with environmental isolates. Geographic trends were identified including regional linkage of several isolate clusters. Multi-locus Sequence Typing (MLST) performed poorly and provided no new insights. Genomes were then screened for antimicrobial resistance genes, which were compared to phenotypic resistance profiles. Isolates showed consistent phenotypic resistance to aminoglycosides with variable resistance to novobiocin and tetracycline. In general, identification of antimicrobial resistance genes did not correlate with phenotypic resistance profiles, indicating unknown genetic determinants. A multi-antimicrobial resistance cassette (aminoglycoside, lincosamide and streptogramin) was identified in the chromosome of three genomes, flanked by vestigial phage recombinases. This locus appears to have spread horizontally within discrete S. uberis populations within a 40 km radius (Sassari region). Genomes were screened for putative virulence factors, which identified 16 genes conserved between sheep and cow isolates, with no host-specific genes shared uniformly across all host-specific isolates. Pangenomic analysis was then performed to identify core genes which were putatively surface-exposed, for identification of potential vaccine targets. As all genomes encoded sortase, core genes were screened for the sortase cleavage motif. Of the 1445 core S. uberis genes, 64 were putative sortase substrates and were predominantly adhesins, permeases and peptidases, consistent with compounds found within ruminant milk such as xanthine, fibronectin and lactoferrin. Conclusions This study demonstrated the importance of whole genome sequencing for surveillance of S. uberis and tracking horizontal acquisition of antimicrobial resistance genes, as well as providing insight into genetic determinants of disease, which cannot be inferred from the MLST schemes. Future mastitis surveillance should be informed by genomic analysis.
 
The geographical distribution of C. parvum strains in Poland. Values in brackets indicate at number of detected parasite strains of particular subtypes. The percentage values shown on the map indicate at C. parvum prevalence in particular province. DS Dolnośląskie, KP Kujawsko-Pomorskie, LB Lubelskie, LD Łódzkie, LS Lubuskie, MP Małopolskie, MZ Mazowieckie, OP Opolskie, PK Podkarpackie, PL Podlaskie, PM Pomorskie, SK Świętokrzyskie, SL Śląskie, WM Warmińsko-Mazurskie, WP Wielkopolskie, ZP Zachodniopomorskie, Nd Not determined
The phylogenetic maximum likelihood tree constructed using the nucleotide sequences (294 bp) of the GP60 gene fragment of C. parvum strains detected in cattle. C. hominis was used as an outgroup to root the tree. C. parvum strains detected in cattle from Poland before 2014 are marked by black diamonds
Article
Background Cryptosporidium parvum ( C. parvum ) is a cosmopolitan parasite that infects various livestock animals including cattle. Microsatellite typing tools for identification of C. parvum subtypes are currently employed to better understand the species-specific epidemiology of cattle cryptosporidiosis. The aim of this study was to analyse the population genetics of C. parvum strains infecting cattle and recognise geographical distribution and time-span correlations in subtype prevalence in Poland. In total, 1601 faecal samples were collected from 2014 to 2018 from healthy cattle from dairy, meat and mixed breeds at the age of 1 week to 4 months. The 267 farms visited were randomly selected and represented all Polish provinces. PCR–RFLP based identification of C. parvum at the 18 small subunit ribosomal RNA (SSU rRNA) locus was performed, followed by strain subtyping by GP60-PCR. Results The overall prevalence of C. parvum in Polish cattle was estimated at 6.2% (100/1601). Animals below the age of 1 month were the major host for this parasite. Excluding one breed, that of dairy-meat mixed, there were no significant differences observed between breed and presence of C. parvum infections (95% TPI All breeds : 1.67–73.53%; POPR = 0.05—0.95). Infected animals were detected in 15 out of 16 Polish provinces, with significant regional prevalence diffrences (Kruskal–Wallis rank sum test, Kruskal–Wallis χ ² = 13.46, p < 0.001). When the population genetics of C. parvum strains were analysed, 11 parasite subtypes from the IIa and IId genetic families were identified. Compared to other parasite strains, IIaA17G1R1 and IIaA17G2R1 appeared at statistically significantly higher frequency (F-test, F = 3.39; p = 0.0003). The prevalence of C. parvum subtypes in cattle was breed-related (Chi-squared test, χ ² = 143.6; p < 0.001). Conclusions The analysis of the population genetics of C. parvum subtypes showed that strains from the IIa subtype family predominated in the tested cattle population. However, relations in changes of subtype prevalence and circulation over time were observed. They were associated with the disappearance of some strains and emergence of new variants from the same genetic family in different geographical locations.
 
Flowchart of the systematic review
(continued)
Main findings and seroprevalence and M. tuberculosis infection prevalence of included studies using other tests
Article
Background Mycobacterium tuberculosis complex (MTC) that causes the chronic infectious disease- tuberculosis (TB), often presents with a complicated epidemiological pattern where the transmission chain may include humans, domestic animals and wildlife, including elephants. TB has been reported globally in both captive and wild elephants. The One Health approach might be the most effective way of understanding the shared MTC infection dynamics in captive and wild animals like Asian elephants. This systematic review accumulates evidence on occurrence, transmission pathways, and preventive measures of TB in elephants from a One Health perspective. Results The prevalence of TB reported in elephant populations ranges from 0 to 23.33% and high prevalence’s are reported for elephants that are in close proximity to infected humans. The risk of elephant to human infection transmission increased significantly with exposure duration and contact with infected elephants. Some studies described the plausible TB transmission to captive elephants from other animals (wild and domestic), suggesting inter- and intra-species transmission. The results of this systematic review based on 27 relevant published works, suggest three overarching interrelated transmission pathways for M. tuberculosis infections in Asian elephants- i) humans and elephants, ii) other animals (wild or domestic) and elephants and iii) unclear sources of infection. Conclusions The progress made with new TB diagnostic tools provides multiple methods to choose from. However, lack of harmonization of TB testing in elephants and their human contacts remains a challenge to prevent TB in those animals. Routine TB screening among elephants and caretakers by setting up an occupational health program for early diagnosis of infection through combined efforts of public health, veterinary medicine, and occupational health experts is suggested. This implies the need for a One Health approach to elephant TB control. This review reveals the need for more research on Mycobacterium tuberculosis complex transmission pathways at the human-animal interface.
 
Right lateral (A), left lateral (B), and ventral-dorsal (C) radiographs of the thorax showing lung hyperinflation and generalized bronchointerstitial pattern with focally increased opacity in the right middle lung lobe and caudodorsal lung fields (white arrowhead)
Pulmonary function assessment by pseudoflow and pseudovolume revealed flow limitation at the late-expiratory phase under natural tidal breathing. Abbreviations: PEF, peak expiratory flow; EF50 and EF25, expiratory flow at 50% and 25% of the remaining tidal volume during tidal breathing
Subpleural ground-glass opacities with distinct margin were observed in the dorsal regions of the bilateral caudal lung lobes (black arrowhead) on lung high-resolution computed tomography (HRCT) (A). At necropsy, the dorsally distributed lesions on lung HRCT corresponded to the multifocal to coalescent dark-red foci (white arrow) on bilateral caudal lobes (B). Histopathology identified the fibrotic narrowing of the bronchioles (C) (Bar scale = 50 μm), and the alveolar septa around these bronchioles was thickened by moderate amount of fibrous connective tissue illustrated using Masson’s Trichrome staining (D) (Bar scale = 500 μm)
Article
Background Bronchiolar disorders are rarely recognized in cats. Constrictive bronchiolitis obliterans is characterized by concentric peribronchiolar fibrosis and inflammation of the bronchioles, but the underlying causes remain poorly understood in current small animal medicine. Case presentation A 9-year-old cat presented with paroxysmal tachypnea, infrequent cough and persistent labor breathing. Thoracic radiography showed lung hyperinflation and bronchointerstitial pattern, and pulmonary function assessment revealed flow limitation in the late-expiratory phase and poor response to short-acting bronchodilator. Dorsally distributed subpleural ground glass opacities with distinct margin and tree-in-bud opacities were observed on lung high-resolution computed tomography. The cat underwent bronchoalveolar lavage (BAL) and showed severe neutrophilic inflammation. Feline herpesvirus was the only pathogen detected in the BAL fluid. Multiple therapeutic attempts were unsuccessful and the cat died 8 weeks after the initial presentation. Necropsy revealed the infiltration of inflammatory cells, obstruction of the bronchiolar lumen, and submucosal concentric fibrosis suggesting constrictive bronchiolitis obliterans. Combining the pre- and post-mortem findings, as well as the time from symptom onset or BAL to necropsy, constrictive bronchiolitis obliterans was possibly triggered by a preceding feline herpesvirus infection in this case. Conclusions The history of nonvaccinated status, lower airway neutrophilic inflammation, and presence of feline herpesvirus in the BAL fluid without coexistence of other pathogens led to the presumption that constrictive bronchiolitis obliterans was induced by a preceding feline herpesvirus infection in this cat. The pathological changes of bronchiolitis obliterans induced by a preceding feline herpesvirus infection could be different from that of cats with acute herpesvirus pneumonia, such as intranuclear inclusions would disappear over time and were no longer found 7–10 days after inoculation. The presence of patchy distribution of subpleural ground glass opacities on lung high-resolution computed tomography should raise the suspicion of peribronchiolar fibrosis. Clinical awareness of bronchiolar disorders as a differential diagnosis is important in cats with lung hyperinflation and labored breathing who show poor reversibility to bronchodilator.
 
Article
Background Repair of large-sized bone defects is a challengeable obstacle in orthopedics and evoked the demand for the development of biomaterials that could induce bone repair in such defects. Recently, UiO-66 has emerged as an attractive metal–organic framework (MOF) nanostructure that is incorporated in biomedical applications due to its biocompatibility, porosity, and stability. In addition, its osteogenic properties have earned a great interest as a promising field of research. Thus, the UiO-66 was prepared in this study and assessed for its potential to stimulate and support osteogenesis in vitro and in vivo in a rabbit femoral condyle defect model. The nanomaterial was fabricated and characterized using x-ray diffraction (XRD) and transmission electron microscopy (TEM). Afterward, in vitro cytotoxicity and hemolysis assays were performed to investigate UiO-66 biocompatibility. Furthermore, the material in vitro capability to upregulate osteoblast marker genes was assessed using qPCR. Next, the in vivo new bone formation potential of the UiO-66 nanomaterial was evaluated after induction of bone defects in rabbit femoral condyles. These defects were left empty or filled with UiO-66 nanomaterial and monitored at weeks 4, 8, and 12 after bone defect induction using x-ray, computed tomography (CT), histological examinations, and qPCR analysis of osteocalcin (OC) and osteopontin (OP) expressions. Results The designed UiO-66 nanomaterial showed excellent cytocompatibility and hemocompatibility and stimulated the in vitro osteoblast functions. The in vivo osteogenesis was enhanced in the UiO-66 treated group compared to the control group, whereas evidence of healing of the treated bone defects was observed grossly and histologically. Interestingly, UiO-66 implanted defects displayed a significant osteoid tissue and collagen deposition compared to control defects. Moreover, the UiO-66 nanomaterial demonstrated the potential to upregulate OC and OP in vivo. Conclusions The UiO-66 nanomaterial implantation possesses a stimulatory impact on the healing process of critical-sized bone defects indicating that UiO-66 is a promising biomaterial for application in bone tissue engineering.
 
Time-dependent antibody-binding activity of beads coupled with LPS from Salmonella serogroup B and C1. Antigen-coupled beads were tested with the Quality Control (QC) panel after 1 day and then monthly for one year. Binding of antibody to beads was calculated as an S/P% value. The plot shows mean S/P% values ± STD for each of the four serogroup B positive sera (S. Typhimurium) and the two serogroup C1 sera (S. Infantis and S. Choleraesuis)
Receiver Operating Curves (ROCs) for Salmonella serogroup B and C1 multiplex-analysis with Area Under Curve (AUC) shown in each graph. The reference assay was an in-house Salmonella mix ELISA
Dot plots with results from the validation of the Salmonella serogroup B and C1 MFIA. Reactivity with antibodies in serum samples from Salmonella mix ELISA-negative pigs (left) and Salmonella mix ELISA-positive pigs (right). Red horizontal lines indicate the cut-off values at the optimal Differential Positive Rate and blue horizontal lines show adjusted cut-off values. Below each graph are shown specificities (Sp) and sensitivities (Se) at the specified cut-off. Levels of antibodies in serum samples is expressed as a percent sample-to-positive ratio S/P%
Percentages of 1425 swine samples from 105 herds that tested negative (grey) or positive (blue) in MFIA and mix ELISA for antibodies to Salmonella enterica serogroup B and C1 in herds previously categorized with a status as Salmonella free (A-D) or Salmonella infected (E–H). Since MFIA distinguishes between serogroup B and C1, percentage of pigs or herds infected with the different serogroups are indicated with blue text
Article
Background Since 1995, a surveillance program for Salmonella has been applied in the Danish pig industry in order to reduce cases of human salmonellosis. The objective of this study was to develop a bead-based Multiplexed Fluorometric ImmunoAssay (MFIA) as an improved serological surveillance method compared to the Salmonella mix ELISA, which has been the national reference immunoassay in the Danish Salmonella surveillance program for about 20 years. Results An MFIA for detection of antibodies to Salmonella serogroup B and C 1 was developed and optimized with regard to coupling of beads with Salmonella lipopolysaccharide antigens and establishing suitable assay conditions. The Salmonella MFIA was validated by testing sera from experimentally infected pigs as well as field sera from non-infected and infected pig herds, and by comparing to results from the Salmonella mix ELISA, which was run in parallel. Sensitivity and specificity was evaluated using receiver operating curve analysis showing an area under curve for the serogroup B and C 1 MFIA of 0.984 and 0.998, respectively. The Salmonella MFIA was shown to detect more antibody-positive samples in seropositive herds compared to the Salmonella mix ELISA, and Bayesian statistics confirmed that the MFIA had a considerably higher sensitivity (94.5%) compared to the mix ELISA (75.1%). The assay specificity was slightly lower for the Salmonella MFIA (96.8%) compared to Salmonella mix ELISA (99.5%). Coupled beads were stable for at least 1 year at 4˚C, and MFIA reproducibility and repeatability of the Salmonella MFIA were acceptable. Results from proficiency tests also indicated that the Salmonella MFIA was more sensitive than the Salmonella mix ELISA and that they had similar specificity. Conclusions A bead-based MFIA for simultaneous detection of porcine serum antibodies to Salmonella enterica serogroup B and C 1 was developed and implemented in the Danish porcine serological Salmonella surveillance program in 2018. The Salmonella MFIA can distinguish, as opposed to the Salmonella mix ELISA, between antibodies to serogroup B and C 1 and the MFIA shows considerably better sensitivity.
 
Pepsin digestion method. A Anesthesia and euthanization of BALB/c mice using the standard laboratory methods, (B) Tissue digestion by digestive solution (pepsin, HCl, and water), (C) Complete digestion by magnetic stirrer, and (D) The modified Baermann technique for the recovery and counting of larvae
DNA amplification of Toxocara cati recovered from BALB/c mice by electrophoresis on 1% agarose gels in the analysis of PCR products. A Infected groups, lane M: 100 bp DNA size marker; lanes 1–5: 600 bp DNA from the liver, lungs, heart, kidneys, and brain respectively. Full-length gel image is provided in Supplementary Fig. 2A. B: Control group, lane M: 100 bp ladder; lanes 1–5: the tissue samples without DNA; lane 6: the negative control; and lane 7: DNA standard. Full-length gel image is presented in Supplementary Fig. 2B
Article
Background Toxocara cati , the cat roundworm, is a parasitic nematode that known to cause toxocariasis in intermediate hosts and humans. In this study, we characterized the dynamics of T. cati larvae migration in BALB/c mice after inoculation with eggs and ensured the migration detecting the larval DNA by a PCR. To evaluate the dynamics of larval migration and distribution, twenty-four BALB/c mice were orally inoculated with 2500 T . cati infective eggs and the visceral organs of the infected animals were examined by pepsin digestion and microscopic parasite counts, followed by PCR at day 1 to 28 post-inoculation. Results The PCR assays were successfully used for detection of T. cati larvae in tissue samples and T. cati larvae and the DNAs were found in the liver, lungs, heart, kidneys and the brain. We detected T. cati in 92.2% of tissue samples by PCR, 30% higher than the conventional pepsin digestion technique. Conclusion Our findings demonstrated that the PCR assay is a sensitive and specific for the detection of T. cati larvae. Therefore, it could become a useful tool for the investigation of the dynamics of larval migration and Toxocara infection in murine model.
 
Comparison of serum ESM-1 concentration in control, MMVD, MMVD substages, death, and alive group. A No significant difference in serum ESM-1 concentration between the control group and substages of MMVD, B ESM-1 concentration was significantly higher in the death group compared to the alive group in MMVD dogs. Abbreviations: ESM-1, endothelial cell-specific molecule-1; MMVD, myxomatous mitral valve disease
Comparison of serum ESM-1 concentration in control, MMVD group with or without comorbidity. No significant difference was found between patients without comorbidities and patients with comorbidities. Abbreviations: ESM-1, endothelial cell-specific molecule-1; MMVD, myxomatous mitral valve disease
Factors affecting serum ESM-1 concentration. A The hypertensive group showed significantly high ESM-1 concentration compared to the normotensive group (p < 0.01). Unit of pressure is mmHg in the table. SBP was classified into normotensive (< 140 mmHg), prehypertensive (140 ~ 159 mmHg), and hypertensive (≥ 160 mmHg) according to ACVIM hypertension consensus (2018), B When classified as VHS, group above 11.5 v showed significantly increased ESM-1 concentration compared to group under 11.5 v (p < 0.05). VHS 11.5, which is considered as general breed cardiomegaly, was used as a cut-off point. C Group taking sildenafil showed significantly high ESM-1 concentration compared to a group not taking sildenafil (p < 0.05). Abbreviations: ESM-1, endothelial cell-specific molecule-1; VHS, vertebral heart score
Change in serum ESM-1 concentration before and after the drug change. Serum ESM-1 concentrations decreased and clinical signs resolved in all patients with increased cardiovascular drug dose (n = 5) (p = 0.0695). Abbreviations: ESM-1, endothelial cell-specific molecule-1
Article
Background Endothelial cell-specific molecule-1 (ESM-1) has emerged as a potential biomarker for cardiovascular disease in humans. Myxomatous mitral valve disease (MMVD) is the most common heart disease in dogs, and we hypothesized that MMVD causes chronic inflammation that increases susceptibility to endothelial glycocalyx (eGCX) damage. In this study, we measured the concentration of ESM-1 in a group of dogs with MMVD and evaluated factors affecting eGCX damage. Results Sixty-four dogs (control, n = 6; MMVD, n = 58) were enrolled in this study. There was no significant difference in serum ESM-1 concentrations among the MMVD stages. The serum ESM-1 concentration was significantly higher in the death group than in the alive group in MMVD dogs. ( p = 0.006). In five dogs with MMVD, serum ESM-1 concentrations tended to decrease when the cardiac drug (pimobendan, furosemide, and digoxin) dose was increased. Conclusions In cases where MMVD progressed to decompensated heart failure with clinical symptoms and resulted in death, the concentration of serum ESM-1 increased significantly. Therefore, ESM-1 could be utilized as a new potential negative prognostic factor in patients with MMVD.
 
Top-cited authors
Holger Andreas Volk
  • University of Veterinary Medicine Hannover
Ramón A Juste
  • Neiker - Basque Institute for Agricultural Research and Development
Andrea Tipold
  • University of Veterinary Medicine Hannover
Clare Rusbridge
  • University of Surrey
Akos Pakozdy
  • University of Veterinary Medicine, Vienna