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Small ruminant lentiviruses (SRLVs) are a group of retroviruses that cause multisystem chronic diseases in goats and sheep and lead to production losses in these animals, negatively affecting animal health and welfare. Although molecular characterization of SRLV field isolates has been performed in many countries, there is currently no information on SRLV genotypes circulating in sheep and goats in Romania. Therefore, the main objective of this study was to conduct a molecular and phylogenetic analysis of SRLVs from Romania and determine the degree of genetic relatedness of the obtained sequences to other known SRLV reference strains. A total of 81 sheep lung tissue samples and 41 sheep lung lymph node samples were tested using nested real-time PCR, and samples positive for real-time PCR were used to amplify an 800 bp gag-pol fragment and an overlapping 625 bp fragment of the gag gene. Pairwise DNA distance and phylogenetic analysis showed that the Romanian SRLV strains were closely related to the A2 and A3 strains based on gag-pol sequences and to the A3 and A17 subtypes based on gag sequences. No recombination events were found. Our results revealed that the Romanian sequences have similar epitope patterns to other existing subtypes, although E/K and R/K mutations in epitope 3 were found only in the Romanian sequences, which may have potential value in serological diagnosis. This study is the first report on the genetic characterization of SRLV strains circulating in Romania and provides new information on SRLV heterogeneity. Further detailed studies should be conducted to better understand the divergence of SRLV Romanian strains.

Background Ovine pulmonary adenocarcinoma (OPA) is caused by Jaagsiekte sheep retrovirus (JSRV) and is considered an important potential animal model for human lung cancer. The precise mechanisms of OPA oncogenesis are still uncertain. The transcription factor signal transducer and activator of transcription 3 (STAT3) is activated by interleukin-6 (IL-6) in many cancers, but this aspect is unknown in OPA. We therefore aimed to evaluate the expression of IL-6 and STAT3 in OPA for its potential role in pulmonary carcinogenesis. Results Lung tissues from 9 grossly normal and JRSV-negative sheep and 20 cases of JSRV-positive OPA sheep were included in the study. Tissue samples were stained with antibodies against IL-6, STAT3, and JSRV-MA. IL-6 and STAT3 were further quantified in both groups using Western Blot (WB). Immunohistochemically, IL‑6 was expressed in stromal, inflammatory, and epithelial cells in all cases of OPA, while STAT3 immunoexpression was restricted to epithelial cells. In the OPA group, the percentage of immunolabelled cells for STAT3 accounted for a mean value of 96%. Using the H-SCORE method, 95% of cases were considered positive for STAT3 expression. Control tissues showed multifocal and weak immunoexpression for both markers. Using WB analyses, a highly significant amount of both IL-6 ( p  = 0.0078) and STAT3 ( p  < 0.0001) proteins were present in lung neoplasms, by comparison to the control lungs. Conclusions Our data showed overexpression of IL-6 and STAT3 in lung tissues from OPA compared to lungs from JSRV-negative sheep. These results suggest a potential role of IL6-STAT3 in OPA carcinogenesis.

Background Angiostrongylus vasorum , commonly known as the “French heartworm,” is a nematode belonging to the Metastrongyloidea superfamily. This parasite was first identified in Toulouse, France in 1853 infecting the pulmonary arteries and the right side of the heart of a Pointer dog. Angiostrongylosis is an important infection due its severe clinical signs and potential for causing high morbidity and mortality in domestic dogs. This nematode has not been studied in Algeria. The aim of this study was investigate the presence of lungworms among different mammal species in a number of Algerian regions. Methods Between February 2022 and September 2023, 47 road-killed animals were collected from six administrative units (departments) in Algeria. All carcasses underwent a full parasitological necropsy, and lung tissues were preserved in 10% buffered formalin and concentrated ethanol for further study. All collected samples were subjected to histological and PCR (cytochrome c oxidase subunit 1 gene) analyses for lungworm identification. Results Histological examination revealed the presence of nematode eggs and larvae in the alveolar space and chronic obstructive vascular changes were detected in a single golden African wolf ( Canis lupaster ) collected from the department of Constantine. First-stage larvae were collected and morphologically identified as Angiostrongylus spp. The molecular identification confirmed the presence of A. vasorum . All other animals tested were negative for lungworms. Conclusions To the best of our knowledge, this is the first report of A. vasorum infection in an African golden wolf ( Canis lupaster ). We report a new host association, highlighting the importance of further studies to update the geographical distribution of A. vasorum and its epidemiology across Algeria. Graphical Abstract

Background: Taraxacum officinale (TO) or dandelion has been frequently used to prevent or treat different liver diseases because of its rich composition in phytochemicals with demonstrated effect against hepatic injuries. This study aimed to investigate the possible preventing effect of ethanolic TO root extract (TOERE) on a rat experimental acute on chronic liver failure (ACLF) model. Methods: Chronic liver failure (CLF) was induced by human serum albumin, and ACLF was induced in CLF by D-galactosamine and lipopolysaccharide (D-Gal-LPS). Five groups (n = 5) of male Wistar rats (200–250 g) were used: ACLF, ACLF-silymarin (200 mg/kg b.w./day), three ACLF-TO administered in three doses (200 mg, 100 mg, 50 mg/kg b.w./day). Results: The in vivo results showed that treatment with TOERE administered in three chosen doses before ACLF induction reduced serum liver injury markers (AST, ALT, ALP, GGT, total bilirubin), renal tests (creatinine, urea), and oxidative stress tests (TOS, OSI, MDA, NO, 3NT). Histopathologically, TOERE diminished the level of liver tissue injury and 3NT immunoexpression. Conclusions: This paper indicated oxidative stress reduction as possible mechanisms for the hepatoprotective effect of TOERE in ACLF and provided evidence for the preventive treatment.

The glycation process is involved in both the intrinsic (individual, genetic) and extrinsic (ultraviolet light, polution and lifestyle) aging processes, and can be quantified at the epidermal or dermal level by histological, immunohistochemical (IHC), or imagistic methods. Our study is focused on a histological and immunohistological comparison of sun-protected regions versus sun-exposed regions from different age groups of skin phototype III subjects, related to the aging process. Skin samples collected from non-protected and UV protected regions of four experimental groups with different ages, were studied using histology and IHC methods for AGE-CML [N(epsilon)-(carboxymethyl)lysine]. A semi-quantitative assessment of the CML expression in the microvascular endothelium and dermal fibroblasts was performed. The Pearson one-way ANOVA was used to compare data between the groups. In the dermis of sun-exposed skin, the number and the intensity of CML positive cells in both fibroblasts and endothelial cells (p<0.05) was higher compared to sun-protected skin, and was significantly increased in older patients. The sun-exposed areas had a more than 10% higher AGE-CML score than the protected areas. No statistically significant correlation was observed between the histological score and the IHC expression of CML. We concluded that in healthy integument, the accumulation of final glycation products increases with age and is amplified by ultraviolet exposure. The study provides new knowledge on differences of AGE-CML between age groups and protected and unprotected areas and emphasizes that endothelium and perivascular area are most affected, justifying combined topical and systemic therapies.

Canine extraskeletal osteosarcomas are mesenchymal, osteoid producing tumors that can arise in soft tissues without initial involvement of the bones. An 8-year-old intact male Beagle dog presented with anorexia, abdominal pain, intermittent vomiting and melena. The patient had a history of recurrent ingestion of cotton based-toy fragments, but no prior surgical procedures involving the abdominal cavity. During the exploratory laparotomy, a mass was identified in the jejunal wall. Surgical resection was performed, and tissue samples were collected for pathological examination. Histologically, the mass was diagnosed as osteoblastic osteosarcoma with fragments of cotton fiber material. The neoplastic cells were immunolabeled for vimentin and BMP-2, further supporting the morphological diagnosis. Seven months after the surgery, metastatic nodules were identified in the liver. The dog died ten months after intestinal mass resection. This case represents the first documented instance of metastatic intestinal osteosarcoma potentially caused by ingestion of cotton fiber material.

Equine herpesviruses are important pathogens causing significant economic loss in equine and asinine populations. EHV-1/4 strains are mainly associated with respiratory distress. The aim of this study is to report the first EHV 4-associated respiratory disease in donkeys in Romania. Thirty-seven of three hundred jennies in an ecological donkey farm in southwest Romania started initially showing signs of severe upper respiratory tract disease, with ten concomitant late abortions/neonatal deaths and three neurological cases. There were nine fatalities. Pathological examination was performed, and samples were collected for Real-Time PCR analysis and histology. In addition, serum samples from 28 individuals with respiratory symptoms were collected and tested using indirect ELISA. RT-PCR identified the EHV-4 strain. Acute, diffuse necrotizing bronchointerstitial pneumonia with occasional intraepithelial intranuclear viral inclusion bodies was identified. Additionally, EHV-1/4-specific antibodies were found in 15 of the 28 sampled animals. Few studies on donkeys and herpesviruses have been published, and this is the first reported case of EHV-4 outbreak in Romania. There is a need for more extensive seroprevalence studies as, currently, the status of EHV-4 infection in donkeys in Romania is unknown.

Background Babesia spp. are apicomplexan parasites which infect a wide range of mammalian hosts. Historically, most Babesia species were described based on the assumed host specificity and morphological features of the intraerythrocytic stages. New DNA-based approaches challenge the traditional species concept and host specificity in Babesia . Using such tools, the presence of Babesia DNA was reported in non-specific mammalian hosts, including B. canis in feces and tissues of insectivorous bats, opening questions on alternative transmission routes. The aim of the present study was to evaluate if B. canis DNA can be detected in tissues of laboratory rodents following oral inoculation with infected ticks. Methods Seventy-five questing adult Dermacentor reticulatus ticks were longitudinally cut in two halves and pooled. Each pool consisted of halves of 5 ticks, resulting in two analogous sets. One pool set ( n  = 15) served for DNA extraction, while the other set ( n  = 15) was used for oral inoculation of experimental animals ( Mus musculus , line CD-1 and Meriones unguiculatus ). Blood was collected three times during the experiment (before the inoculation, at 14 days post-inoculation and at 30 days post-inoculation). All animals were euthanized 30 days post-inoculation. At necropsy, half of the heart, lung, liver, spleen and kidneys were collected from each animal. The presence of Babesia DNA targeting the 18S rRNA gene was evaluated from blood and tissues samples. For histopathology, the other halves of the tissues were used. Stained blood smears were used for the light microscopy detection of Babesia . Results From the 15 pools of D. reticulatus used for the oral inoculation, six were PCR-positive for B. canis . DNA of B. canis was detected in blood and tissues of 33.3% of the animals (4 out of 12) inoculated with a B. canis- positive pool. No Babesia DNA was detected in the other 18 animals which received B. canis -negative tick pools. No Babesia was detected during the histological examination and all blood smears were microscopically negative. Conclusions Our findings demonstrate that B. canis DNA can be detected in tissues of mammalian hosts following ingestion of infected ticks and opens the question of alternative transmission routes for piroplasms.

The aim of the present study was to determine and compare the degree and duration of corneal anesthesia following topical application of 0.4% oxybuprocaine hydrochloride ophthalmic solution and 1% ropivacaine hydrochloride treatment in healthy rats. A randomized, blinded, crossover study was conducted on 20 healthy adult Wistar rats, following complete physical and ophthalmological examination. Baseline corneal touch threshold (CTT) was determined in the central corneal area of both eyes with a Cochet–Bonnet aesthesiometer, in mm filament length. Oxybuprocaine was randomly applied to one eye and 0.9% sterile sodium chloride solution was instilled into the contralateral eye. Subsequent CTT measurements were performed in both eyes 5 minutes after topical application and at 5-minute intervals thereafter for 75-minutes in the anesthetized eye. Following a 2-week washout period, this protocol was repeated with ropivacaine. Quantitative data were summarized as mean ± standard deviation, median and inter-quartile range (Q1–Q3). Repeated measures data were analyzed over time and between treatments using Friedman test and Wilcoxon signed-rank test with Bonferroni adjustment ( p < 0.05). Baseline CTT values were 60 mm in all eyes. With oxybuprocaine, CTT values decreased significantly for 65 minutes (0–55 mm; p = 0.002) when compared with baseline; the maximal anesthetic effect (no blink response at 5 mm filament length) was maintained for up to 15 minutes ( p < 0.0001). With ropivacaine, CTT values were significantly lower than baseline for 30 minutes (0–55 mm; p = 0.002), with a maximal anesthetic effect recorded at 5 minutes in 18 eyes ( p < 0.0001). Oxybuprocaine induced a significantly lower CTT than ropivacaine ( p = 0.002) from 10 to 65 minutes following topical application. Both anesthetic agents induced significant corneal anesthesia; however, oxybuprocaine provided a greater and longer anesthetic effect, making it more suitable for potentially painful ophthalmologic procedures.

This report provides a clinical, histological, and immunohistochemical description of an unusual hibernoma (pale cell variant) in the subepidermal area of the nipple of a six-year-old bitch. Furthermore, an extensive literature review of hibernomas in animals was made. Physical examination revealed a nodular lesion in the subepidermal area of the third nipple of the left mammary chain. The histopathological findings included lobules of round to oval cells with abundant pale to eosinophilic cytoplasm, containing one or multiple optically empty vacuoles, consistent with nipple hibernoma. Immunohistochemically, the neoplastic cells were negative for cytokeratin AE1/AE3 and p53 but showed strong immunoreaction for vimentin and uncoupling protein-1, thus confirming the brown adipose tissue origin. Local recurrence was not detected after 18 months of follow-up. Hibernomas are rare and benign neoplastic lesions, originating from brown adipose tissue. Due to their histological and molecular resemblance with liposarcoma, a correct diagnosis of these neoplasms is required. In addition, the literature review suggests that hibernomas may present different features, according to species.