Experimenting with Pig-based Skin Model for Burns. Testing of Mean Literature Findings.

Introduction: Animal models that closely mimic human burn wound healing processes are essential for developing effective burn wound treatments. Pigs are useful animal models for studying burn wound healing. From their extensive literature review, Andrews and Cuttle (2017) reported mean temperature and exposure time values. This study was done to provide initial burn depth for another experiment comparing two burn wound treatments. The secondary goal was to validate a systematic review on porcine burn model standardization. Materials and Methods: Six four-week-old Large White x Landrace gilts were housed in a closed structure for 10 days to acclimatize. The procedures were performed under general anesthesia. A round 2.5 cm copper plate welded to an aluminum rod with a wooden handle caused the injuries. The burning device was used to reach a contact temperature of 110ºC on the pig's skin. The objective was to create a superficial partial thickness (SPT) burn for 10 seconds (Group 10s) and a deep partial thickness (DPT) burn for 20 seconds (Group 20s) using a plate heated at 110ºC. No stabilizer or pressure controller was used. Wounds were conclusively dressed and harvested 24 hours later. The usual hematoxylin-eosin protocol was used to cut and stain 4-micron sections. Results: A significant difference (p 0.01) was observed in dermis involvement, with a mean of 85.61 % (95% CI= 80.62 to 90.61) for group 10s and 123.71% (95% CI= 114.91 to 132.50) for group 20s. An exposure time of 20 seconds increased dermis depth-related total collagen denaturation by almost 50% compared to 10 seconds. Conclusions: In conclusion, our experiment produced DPT burns in 10 seconds and FT burns in 20 seconds without a pressure application device.

Experimenting with Pig-based Skin Model for Burns. Testing of Mean Literature Findings.

Introduction: Animal models that closely mimic human burn wound healing processes are essential for developing effective burn wound treatments. Pigs are useful animal models for studying burn wound healing. From their extensive literature review, Andrews and Cuttle (2017) reported mean temperature and exposure time values. This study was done to provide initial burn depth for another experiment comparing two burn wound treatments. The secondary goal was to validate a systematic review on porcine burn model standardization. Material and Methods: Six four-week-old Large White x Landrace gilts were housed in a closed structure for 10 days to acclimatize. The procedures were performed under general anesthesia. A round 2.5 cm copper plate welded to an aluminum rod with a wooden handle caused the injuries. The burning device was used to reach a contact temperature of 110 C on the pig's skin. The objective was to create a superficial partial thickness (SPT) burn for 10 seconds (Group 10s) and a deep partial thickness (DPT) burn for 20 seconds (Group 20s) using a plate heated at 110ºC. No stabilizer or pressure controller was used. Wounds were conclusively dressed and harvested 24 hours later. The usual hematoxylin-eosin protocol was used to cut and stain 4-micron sections. Results: A significant difference (p 0.01) was observed in dermis involvement, with a mean of 85.61 % (95% CI= 80.62 to 90.61) for group 10s and 123.71% (95% CI= 114.91 to 132.50) for group 20s. An exposure time of 20 seconds increased dermis depth-related total collagen denaturation by almost 50% compared to 10 seconds. Conclusions: In conclusion, our experiment produced DPT burns in 10 seconds and FT burns in 20 seconds without a pressure application device.

Effects of age and season on conception rate of Mediterranean Italian Dairy Buffalo (Bubalus bubalis) following oestrus synchronization and fixed-time artificial insemination.

The objective of this project was to report the effects of season and age on conception rate amongst Italian Mediterranean Buffalo subjected to an Ovsynch/Resynch (O/R) reproductive management protocol. The study utilized nulliparous (heifers), primiparous buffalo cows (PBC) and multiparous buffalo cows (MBC). The primiparous and multiparous groups were subjected to the synchronization protocol throughout the entire year, but heifers were synchronized and inseminated only during the spring/summer seasons. The conception rate obtained following the OvSynch oestrus synchronization protocol, applied during spring, was 68.4% for heifers, 83.3% for PBC and 67.7% for MBC. The overall total conception rates following the complete O/R protocol were 84.27%, 94.4% and 79%. Conception rates achieved during summer were heifers 52%, PBC 47.2% and MBC 49%, whilst overall conception rates following the full O/R protocol were 72%, 69.8% and 58.2% respectively. In the autumn seasons, PBC conceived 58.9% and MBC 52.1% following initial Ovsynch, which improved to total overall conception rates of 87.5% and 78.7% following the full O/R protocol. Similarly, in the winter season, PBC experienced a conception rate of 47.5% following Ovsynch and 72.5% after a follow-up Resynch. MBC experienced 60.0% and 74.4% conception following Ovsynch and full O/R, respectively, during winter. Total conception rates during all seasons were quite acceptable following the O/R protocol. There was a significant decrease from spring to summer in conception rate for all parity groups, but heifers were not a severely affected as older buffalo cows. This finding agrees with that of other investigators indicating that heifer fertility is not as negatively impacted by long photoperiod and higher ambient temperature as that of older animals. The O/R protocol as utilized in this study is an effective means of reproductive management for dairy buffalo cows and is effective for improving fertility during out-of-season breeding.

Utilization of Rosmarinic and Ascorbic Acids for Maturation Culture Media in Order to Increase Sow Oocyte Quality Prior to IVF.

The beneficial effect of antioxidant supplementation in maturation culture media of sow oocytes was evaluated by the expression quantification of apoptotic genes and the genes that ensure stability of germ cells during fertilization. The oocytes were cultivated for 44 h in conventional medium (C) or in medium supplemented with 105 µM rosmarinic acid (R) and 0.5 mM ascorbic acid (A) and classified into three quality classes by morphological observation from which the total RNA was isolated. The gene expression of Ptx3 and the apoptotic regulator p53, Bax and BCL-2 were evaluated by quantitative PCR technique. The decreased expression of the Bax gene in the A and R groups, compared to the control, indicates a protective role of antioxidants in the cells. Cell homeostasis was maintained, as reflected in the ratio of Bax/Bcl-2 in class I COCs (cumulus-oocyte complex) regardless of the experimental group, indicating minimum cellular stress. The expression of p53 genes was higher in all class III COC, but in A1 and R1 the expression was lower than in C1, and a similar Ptx-3 gene decreased significantly in groups A1, A2, A3 and R1 compared with control groups. Antioxidant supplementation showed beneficial effects on all morphological classes of pig COCs.

Early sacral neuromodulation ameliorates urinary bladder function and structure in complete spinal cord injury minipigs.

AIMS: To determine the effects of early sacral neuromodulation (SNM) and pudendal neuromodulation (PNM) on lower urinary tract (LUT) function, minipigs with complete spinal cord injury (cSCI) were analyzed. SNM and PNM have been proposed as therapeutic approaches to improve bladder function, for example after cSCI. However, further evidence on efficacy is required before these methods can become clinical practice. METHODS: Eleven adults, female Göttingen minipigs with cSCI at vertebral level T11-T12 were included: SNM (n = 4), PNM (n = 4), and SCI control (SCIC: n = 3). Tissue from six healthy minipigs was used for structural comparisons. Stimulation was started 1 week after cSCI. Awake urodynamics was performed on a weekly basis. After 16 weeks follow-up, samples from the urinary bladder were taken for analyses. RESULTS: SNM improved bladder function with better capacities and lower detrusor pressures at voiding and avoided the emergence of detrusor sphincter dyssynergia (DSD). PNM and untreated SCI minipigs had less favorable outcomes with either DSD or constant urinary retention. Structural results revealed SCI-typical fibrotic alterations in all cSCI minipigs. However, SNM showed a better-balanced distribution of smooth muscle to connective tissue with a trend towards the reduced progression of bladder wall scarring. CONCLUSION: Early SNM led to an avoidance of the emergence of DSD showing a more physiological bladder function during a 4 month follow-up period after cSCI. This study might pave the way for the clinical continuation of early SNM for the treatment of neurogenic LUT dysfunction after SCI.

A new technique for minimal invasive complete spinal cord injury in minipigs.

BACKGROUND: The aim of this study was to develop a minimal invasive complete spinal cord injury (SCI) minipig model for future research applications. The minipig is considered a translationally relevant model for SCI research. However, a standardized minimal invasive complete SCI model for pigs has not yet been established. METHODS: Adult Göttingen minipigs were anesthetized and placed in extended prone position. After initial computed tomography (CT) scan, the skin was incised, a needle placed in the epidural fatty tissue. Using the Seldinger technique, a guidewire and dilators were introduced to insert the balloon catheter to Th12. After confirmation of the level Th11/Th12, the balloon was inflated to 2 atm for 30 min. The severity of the lesion was followed by CT and by MRI, and by immunohistochemistry. Function was assessed at the motor and sensory level. RESULTS: Duration of procedure was about 60 min including the 30-min compression time. The balloon pressure of 2 atm was maintained without losses. The lesion site was clearly discernible and no intradural bleeding was observed by CT. Neurological assessments during the 4-month follow-up time showed consistent, predictable, and stable neurological deficits. Magnetic resonance imaging analyses at 6 h and 4 weeks post SCI with final immunohistochemical analyses of spinal cord tissue underlined the neurological outcomes and proved SCI completeness. CONCLUSIONS: We have established a new, minimal invasive, highly standardized, CT-guided spinal cord injury procedure for minipigs. All risks of the open surgery can be excluded using this technique. This CT-guided SC compression is an excellent technique as it avoids long surgery and extensive trauma and allows a feasible inter-animal comparison.

Serum Protein Electrophoretic Pattern in Neonatal Calves Treated with Clinoptilolite.

The objective of our study was to determine the effects of clinoptilolite supplemented in colostrum on the blood serum protein electrophoretic pattern of new-born calves. METHODS: Romanian Black and White new-born calves involved in the study were divided into 3 groups: the control group (C) that received colostrum without clinoptilolite, and experimental groups I (E1) and II (E2) that received colostrum supplemented with 0.5% and 2% clinoptilolite, respectively. The concentration of total protein and protein fractions (albumin, α1-globulin, α2-globulin, ß-globulin and γ-globulin) were analyzed by electrophoresis on cellulose acetate. RESULTS: At hour 30 after birth, concentrations of γ-globulins, ß-globulin and total protein in E1 group of calves were higher than in control group by 42.11% (p < 0.05), 28.48% (p > 0.05) and 18.52% (p > 0.05), respectively, and were higher, but not significantly, in group E2 compared to the control group. This was in accordance with a significant lower albumin/globulin ratio in groups E1 and E2 (29.35%, p < 0.05 and 35.87%, p < 0.05, respectively) than in control group at 30 h postpartum, which indicates an obvious increase of the globulins fraction in experimental groups. The conclusion: Clinoptilolite was effective in improving passive transfer in new-born calves, but it was more effective if added in colostrum with a dose of 0.5% than with a dose of 2%.

Correction: Lungu et al. Molecular Characterisation of Antimicrobial Resistance in E. coli Isolates from Piglets in the West Region of Romania. Antibiotics 2023, 12, 1544.

The authors Bianca Cornelia Lungu and Ioan Hutu did not state contributed equally [...].

Microbiological and Molecular Investigation of Antimicrobial Resistance in Staphylococcus aureus Isolates from Western Romanian Dairy Farms: An Epidemiological Approach.

Antimicrobial therapy is the most frequently used medical intervention for bovine mastitis in the dairy industry. This study aims to monitor the extent of the antimicrobial resistance (AMR) problem in Staphylococcus aureus in the dairy industry in Western Romania. Twenty farms were selected by random sampling in a transverse epidemiological study conducted across four counties in Western Romania and divided into livestock units. This study assessed the association between the resistance genes to phenotypic expression of resistance and susceptibility. Isolates of S. aureus were identified and q-PCR reactions were used to detect antibiotic resistance genes. One hundred and fifty bovine and 20 human samples were positive for S. aureus. Twenty five percent of bovine isolates (30/120) and none(0/30) of the human isolates were methicillin-resistant S. aureus (MRSA). All isolates were susceptible to fosfomycin, ciprofloxacin, netilmicin, and resistant to ampicillin and penicillin. S. aureus isolates regarded as phenotypically resistant (R) were influenced by the origin of the samples (human versus bovine, χ2 = 36.510, p = 0.013), whether they were methicillin-resistant S. aureus (χ2 = 108.891, p < 0.000), the county (χ2 = 103.282, p < 0.000) and farm of isolation (χ2 = 740.841, p < 0.000), but not by the size of the farm (χ2 = 65.036, p = 0.306). The multiple antibiotic resistance index was calculated for each sample as the number regarded as phenotypically resistant (R)/total antibiotics tested (MARI = 0.590 ± 0.023) was significantly higher (p < 0.000) inmethicillin-resistant S. aureus (0.898 ± 0.019) than non-methicillin-resistant S. aureus (0.524 ± 0.024) isolates. For the antibiotics tested, the total penetrance (P%) of the resistance genes was 59%, 83% for blaZ, 56% for cfr, 50% for erm(B), 53% for erm(C), 57% for mecA and 32% for tet(K). Penetrance can be used as a parameter for guidance towards a more accurate targeting of chemotherapy. P% in S. aureus was strongly positively correlated with the multiple antibiotic resistance index (r = +0.878, p < 0.000) with the potential to use the same limit value as an antibiotic management decision criterion. Considering cow mastitis, the penetrance value combined with the multiple antibiotic resistance index suggests that penetrance could serve as a useful parameter for more precise targeting of chemotherapy for S. aureus.

Diagnostic Pathways of Leptospira spp. in Dogs with Kidney Injury.

Pathogenic Leptospira spp. causes leptospirosis in animals and humans globally, leading to systemic infections that can impact vital organs in affected animals. The purpose of this study was to evaluate kidney injury and to perform a retrospective analysis of leptospirosis infection in follow-up dog samples. The retrospective study collected epidemiological information obtained through paraclinical exams, immunohistochemistry (IHC), and molecular biology (qPCR) of cases from the Faculty of Veterinary Medicine from Timisoara between September 2016 and May 2023. No correlations were found between Leptospira infection and breed (p = 0.714), gender or castration status (p = 0.890), and anatomic pathology exam results (p = 0.608). Significant associations were found in cases with high levels of azotemia (p = 0.000) and immunological status (vaccinated vs. unvaccinated, p = 0.000), with the leptospirosis risk in unvaccinated animals calculated at OR = 85.500 (95%CI, 6.82-1071.26, p = 0.000). Retrospectively, leptospirosis was diagnosed in 27/65 cases (42%) using the IHC method, while the qPCR assay detected 29/65 cases (45%). This study demonstrates that qPCR is a robust and specific method for postmortem diagnosis of Leptospira spp. infection in dogs, offering higher specificity and reliability compared to traditional IHC methods, which showed 94.74% specificity in our study.

Molecular Characterisation of Antimicrobial Resistance in E. coli Isolates from Piglets in the West Region of Romania.

Antibiotics are widely used for prophylaxis and therapy, reducing morbidity and mortality produced by bacterial pathogensin pigs, including infections caused by Escherichia coli. The aim of this study was to characterise antibiotic resistance phenotypes and genotypes in E. coli isolates in pigs in West Romanian grower farms. Differential phenotypic susceptibility profiles and the contribution of resistance genes to phenotypic expression of susceptibility or resistance were evaluated. A total of 76 E. coli isolates were identified and confirmed by the MicroScan Walk Away System. The occurrence of four resistance genes, ampC, blaZ, blaTEM and tetK in strains resistant to 13 antibiotics was assessed. Of the E. coli isolates, 0% showed resistance to meropenem, 3.9% to tigecycline and 10.5% to piperacillin/tazobactam, whereas, in contrast, 100% were resistant to ampicillin and mezlocillin, 76.31% to piperacillin and 59.3% to tetracycline. The prevalence of resistance genes in resistant isolates detected by q-PCR analysis was 97.0% for ampC, 96% for blaZ, 32.9% for blaTEM and 58.8% for tetK. Penetrance (the proportion of individuals carrying a particular variant of a gene that also expresses an associated trait) was 50% for ampC (32% for amoxicillin/clavulanate, 62% for cefazolin, 32% for cefepime, 100% for cefotaxime, 56% for cefuroxime and 99% for ampicillin), 65% for blaZ (32% for amoxicillin/clavulanate and 99% for ampicillin), 51% for blaTEM (81% for piperacillin) and 44% for the tetK gene (83% for tetracycline). The result of phenotypic antibiotic resistance testing may indicate the presence of plasmid-borne resistance, with a diagnostic odds ratio of a positive phenotypic resistance for tetK being 4.52. As a management decision, the maximum penetrance admitted for using a specific antibiotic for E. coli infections in pigs is recommended to be less than 20%.

Unusual Canine Distemper Virus Infection in Captive Raccoons (Procyon lotor).

Canine morbillivirus, also known as canine distemper virus (CDV), is the causative agent of canine distemper (CD), which is a serious contagious disease of canines, large felids, and, occasionally, raccoons. This study included seven raccoons from the Timisoara Zoological Garden, Romania. CDV was detected using RT-qPCR on blood samples, but several other exams were also performed-clinical, bacteriological, immunohistochemistry (IHC) and histopathology, toxicological screening, and necropsy-which confirmed CDV infection. Severe digestive disorders (diarrhea and frequent hematemesis) were observed. The necropsy findings included pseudo membranous gastroenteritis, congestion, and pulmonary edema in two raccoons. Immunohistochemistry showed immunolabeled CDV antigenantibodies on the viral nucleocapsid. Histopathology revealed lymphocyte depletion in mesenteric lymphnodes and intranuclear and intracytoplasmic inclusions in the enterocytes of the small intestine. Based on the RT-qPCR assay, laboratory tests, and the lesions observed, it was established that the raccoons were infected with CDV, which was the cause of death in two cases. The results from the necropsy, histology, and immunohistochemistry in the raccoons are comparable with reported CDV lesions in dogs. In conclusion, several exams may be performed to establish the etiology of possible interspecific viral infection, but only very specific exams can identify aCDV infection. Laboratory analyses must be completed by RT-qPCR assay or IHC to establish infection with uncommon viruses in raccoons with high accuracy.

Structural Changes of the Urinary Bladder After Chronic Complete Spinal Cord Injury in Minipigs.

PURPOSE: The aim of this study was to determine the structural changes of the urinary bladder after chronic spinal cord injury (SCI) in minipigs with the primary focus on the analysis of urinary bladder wall proteins and their quantitative distribution. METHODS: Seven Göttingen minipigs (adult, female) underwent a complete spinal cord transection. Follow-up time was 4 months during which the bladder was drained by frequent single catheterisation and data from the bladder diary and daily urine strip test were collected. Samples from the urinary bladder were taken, fixed in 4% paraformaldehyde and stained for histological analyses. Bladder wall thickness, single tissue quantities/distributions, types I and III collagen, and elastin quantifications were performed. Comparisons to healthy urinary bladder tissue of age-matched minipigs were performed for statistical analyses. RESULTS: No urinary tract infections were observed in our SCI minipig collective during follow-up. A trend towards a reduction in bladder volumes and an increase in incontinence periods were seen. The bladder wall thickness significantly increased after chronic SCI. Furthermore, bladder wall composition was severely altered by a significant loss of smooth muscle tissue and a significant increase in connective tissue. Elastic fibres were reduced in number and altered in their structural appearance after SCI. Type I collagen was significantly increased, while type III collagen was significantly decreased after SCI. CONCLUSIONS: Chronic SCI highlighted that the urinary bladder wall undergoes fibrotic events with reduced contractile and elastic properties due to changes of the bladder wall protein composition. These changes show in detail how SCI severely influences the urinary bladder wall composition and depicts the similarities between minipigs and humans.