The main objectives of this study were to investigate surfactant apoprotein expression (SP) and to detect Brucella sp. antigens in the lungs of aborted bovine fetuses and neonatal calves delivered weak. The Avidin-Biotin-Peroxidase Complex (ABC) and the indirect immunofluorescence (IF) techniques were applied, using antibodies to the lung surfactant apoproteins (SP-A, SP-B, SP-C) and Brucella sp. antigens. Hyperplasia of type II cells was also assessed by evaluating Thyroid Transcription Factor-1 (TTF-1), Proliferating Cell Nuclear Antigen (PCNA), and Cytokeratin Pan Type I/II (CK-P) markers. The study materials were the lungs of 46 aborted bovine fetuses and 20 neonatal calves delivered weak. Brucella sp.-positive fetal lungs displayed bronchopneumonia in 24 cases. The lungs of the weak-delivered neonates which were positive for Brucella sp. also showed pneumonia. Bacterial culture detected positivity in 11 of 46 fetuses and two neonates. IHC for Brucella sp. antigens found positivity in 22 of 46 fetuses and four neonates. Thus, our research revealed that the IHC technique using anti-Brucella sp. antibodies was useful for detecting Brucella sp. in autolytic and culture-negative fetuses. The study also found that surfactant synthesis begins close to the 7th month of gestation in bovine fetuses. Immunolabeling to SPs occurred in the cytoplasm of both type II and Clara cells, along with SP-C only in type II pneumocytes. The IF yielded dense labeling for Brucella sp. antigens, SP-B, and CK-P, respectively, in the phagocytic cells and epithelium of the airways. Also, pneumonia in newborn calves indicates an intrauterine infection by Brucella sp.

Metritis has considerable economic impact on dairy herd profitability due largely to reduced reproductive performance. Studies in Ethiopia only report prevalence, with no available data on the incidence of metritis. The present prospective cohort study aimed to estimate the incidence of puerperal metritis, identify risk factors, and isolate the causative bacteria in dairy cows in Hawassa. Daily follow-up of 120 dairy cows from parturition to 21 days postpartum was carried out from November 2019 to February 2021. Of these, 21 developed puerperal metritis with an incidence rate of 0.94 cases per 100 cow days at risk (95 % CI: 0.6-1.4). The cumulative incidence was found to be 17.5 % (95 % CI: 11.8-25.6). Over 90 % of the puerperal metritis occurred within the first 10 days of parturition. The mean day of occurrence of puerperal metritis was 7.7 days postpartum and the mean rectal temperature in metritic cows was 40.5 °C. Retained fetal membranes (RFM) (p < 0.001) emerged as the only predisposing factor for metritis in the final Cox regression model. Cows with RFM faced a 14.9 times higher risk of metritis compared to cows without RFM. A total of 52 bacterial isolates, belonging to 4 genera, were recovered from the aerobic culture of 21 uterine swab samples. E. coli and Staphylococcus spp. were the most frequently isolated bacteria, with recovery from 21 (100 %) and 20 (95.2 %) of the metritic uteri, respectively. Results of this study suggested the importance of reducing incidence of RFM to reduce incidence of metritis in dairy cows.

Anethole, an antioxidant found in plants, appears to improve the survival of spermatozoa during semen cryopreservation. This study assessed the effects of commercial trans-anethole in ram semen cryopreservation. Thirty ejaculates from six rams were diluted in media containing anethole at the following concentrations: CONT (0 μM), AN10 (10 μM), AN50 (50 μM), and AN100 (100 μM). Semen was slow-frozen, preserved in liquid nitrogen, and thawed. Anethole at 10 μM or 50 μM did not compromise any studied sperm quality parameter but increased pre-freezing functionality of membrane and mitochondrial activity. At 10 μM, anethole reduced post-thawing spermatozoa lipoperoxidation. At 50 μM, anethole sustained higher mitochondrial activity after thawing, reduced minor defects in sperm, and increased the number of sperm binding to perivitelline membrane, while keeping lipoperoxidation levels as in control. Anethole at 100 μM promoted higher pre- and post-freezing mitochondrial activity and higher number of sperm binding to perivitelline membrane, in comparison to control. Additionally, some post-thawing kinematic parameters were enhanced by anethole at 100 μM. Of note, mitochondrial activity and lipoperoxidation were higher with anethole at 100 μM in comparison to 50 μM, not differing from control. At the hypoosmotic test, the highest concentration (100 μM) tested reduced sperm osmotic resistance. The results of this study indicate that using anethole in cryopreservation media promoted mostly positive effects on the fresh and post-thawed ram semen, and the advantages vary according to its concentration.

This systematic review presents scientific evidence on treatment modalities for claw horn lesions (CHLs) in dairy cows, and their efficacy in restoring normal gait, resolution of claw lesions, and improving nociceptive threshold. A literature search was performed in four databases leading to the synthesis of 10 articles for the final analysis. The types of treatment administered include single (STM), bimodal (BTM) and multimodal treatment modalities (MTM). Locomotion scores (LS) were the most reported outcome measure, followed by the resolution of claw lesion, time to lameness cure, nociceptive threshold and gait properties. Most pairwise comparisons for LS and nociceptive threshold depicted no significant difference between the various treatment groups. Meanwhile, a few pairwise comparisons in terms of resolution of claw lesion and time to lameness cure revealed that cows receiving MTM (claw trimming + hoof block + NSAID) or BTM (claw trimming + hoof block) recorded complete resolution of CHL and significantly less time to recover compared to those treated with an STM (only claw trimming). Despite the various modalities used in CHL treatment, their effects on LS, nociceptive threshold, and gait properties are still unclear. While evidence suggests that MTM and BTM facilitate rapid resolution of CHL and duration of lameness cure, only limited data are available and the association with specific lesions is poorly understood. Several sources of heterogeneity were identified between the studies, particularly duration of follow-up after treatment, LS systems, severity and duration of lameness, and types of treatment administered.

Klebsiella pneumoniae (KP) is a zoonotic conditionally pathogenic bacterium with a high prevalence of infection. It often induces purulent inflammation of the rabbit lungs, kidneys and other tissues, with high morbidity and mortality. Bacillus coagulans (BC) has the ability to regulate the balance of host intestinal flora and improve host immunity. However, the mechanism of the protective effect of BC on KP-induced kidney injury in rabbits is not clear. To explore this, we randomly divided fifty 35-day-old Eyplus rabbits into five groups: control, KP, low-dose BC (LBC), medium-dose BC (MBC) and high-dose BC (HBC). On the 1st day of the experiment, rabbits in LBC, MBC and HBC groups were fed diets containing 1 × 10 CFU/g, 5 × 10 CFU/g and 1 × 10 CFU/g BC, respectively, and rabbits in CK and KP groups were fed basal diets. On the 8th day, each rabbit in the KP, LBC, MBC and HBC groups was infused with 4 mL of 1 × 10 CFU/mL KP bacterial solution, and the CK group was infused with the same amount of sterilised saline for a total of 7 days. Rabbit kidney tissues were collected on the 15th d. HE staining was used to observe the pathological changes of rabbit kidney tissues, oxidative stress-related indexes were detected by biochemical kits, and the content of inflammatory factors and apoptosis-related factors in kidney tissues were detected by ELISA. The results showed that KP disrupts the normal structure of the kidney, induces oxidative stress and inflammatory responses, and mediates apoptosis by regulating the levels of Bcl-2 family proteins. BC pretreatment significantly reduced kidney structural damage, oxidative stress, inflammatory response, and apoptosis in rabbits. To alleviate KP-induced kidney injury by increasing the activity of antioxidant enzymes and the content of anti-apoptotic proteins. Compared with the LBC group and the HBC group, the remission effect was more pronounced in the MBC group. Therefore, in this study, the effect of 5 × 10 CFU/g BC was more significant.

The objectives of this study were: (i) to verify and compare the coat surface temperature (CST) of regions of interest (ROI) measured through infrared pyrometer (IRP) and thermography (IRT) under two environmental conditions, to evaluate (ii) reliability and (iii) sensitivity of the two infrared equipments to the environmental effect. Twenty hair ewes (Ovis aries), multiparous, non-lactating, non-pregnant, with body weight 59.38 ± 6.23 kg and body score condition ranging from 2.5 to 4.5 were evaluated. All CST, in both infrared methods, were higher (P < 0.001) in the afternoon (2 to 3 p.m.), with the exception of eyes surface temperature (TSEY) from IRT, which was higher (P < 0.001) in the morning (8 to 9 a.m.). Regarding the infrared methods at different times (twice a day, morning and afternoon), of the day, CST values for all ROIs were significantly higher (P < 0.001) in thermography, regardless of the time shift, except for ear pinna surface temperature (TSE) and rectum surface temperature (TSR), which showed no significant difference (P > 0.05) between the infrared methods during the morning shift. The thermal environment significantly influences (P < 0.001) the surface temperature of various anatomical regions in both infrared methods. The correlation coefficients between IRT and IRP were moderate. The IRT is more sensitive to the thermal environment, as it presented higher canonical coefficients in the environmental variables. Among surface temperatures, eye surface temperature is the most influenced by variables in the thermal environment, especially THI. All three methods (digital thermometer-DT, IRT and IRP) showed >80 % of the data variation in the first two components. Using the DT, we observed an association between vaginal temperature (VT) and RT with air temperature (AT) and temperature humidity index (THI), which was similar to the infrared thermography pattern, in which all ROIs were highly correlated with AT and THI. The opposite was observed in the pyrometer, in which an association between eye temperature and RH and lower factor loadings of the other ROIs with AT and THI in the first component was observed. In conclusion, while IRP and IRT showed similar performance in the morning, IRT proved to be more accurate in the afternoon, demonstrating a higher success rate in classifying the group of origin. Therefore, it is concluded that IRT is more reliable than IRP in detecting surface temperature changes under high temperatures.

Cysticercosis, caused by the larval stage of Taenia solium, is a major zoonotic parasitic disease with a global impact, heightened by immigration and transboundary activities. Recognizing its importance, the World Health Organization has prioritized eliminating taeniasis/cysticercosis by 2030, designating it as a priority neglected tropical disease (NTD). Accurate diagnosis is essential for effective disease control, yet traditional methods often fall short due to low sensitivity of the test and the need for specialized laboratory setups. This study focuses on developing a recombinant Ag2 (rAg2)-based dot blot assay to detect T. solium cysticercosis in pigs. The rAg2 antigen was successfully expressed and purified, with immunoreactivity confirmed against sera from naturally infected pigs. The dot blot test evaluated against serological gold-standard enzyme-linked immunotransfer blot (EITB) assay, demonstrated preliminary laboratory sensitivity (89 %, 95 % CI: 68.6-97.1) and specificity (100 %, 95 % CI: 97.6 - 100), highlighting its potential as a reliable alternative diagnostic method. Cost-effectiveness and minimal laboratory requirements make it particularly suited for field use in resource-limited settings.

Cannabidiol (CBD) has been shown to have anti-inflammatory and antipruritic properties without the significant psychoactive effects. This study aims to evaluate the cytotoxic effects of, and the production of cytokines after exposure to CBD, cannabidiolic acid (CBDA), and β-caryophyllene (BCP), alone and in combination, by peripheral blood mononuclear cells (PBMC) from healthy and atopic dogs. Six healthy and five atopic, privately-owned dogs were enrolled. Peripheral blood mononuclear cells were harvested and incubated for 24 h with different concentrations of CBD, CBDA, and BCP alone or in combination. Cell viability and inflammatory cytokines were assessed. There was no difference in cell viability between baseline and tested concentrations of CBD, CBDA, or BCP in either healthy or in atopic PBMC. There was no effect of CBD, CBDA and BCP on the secretion of cytokines compared to baseline in healthy or atopic PMBC. The only exception was interleukin (IL)-10, increased in healthy PMBC exposed to CBD 100 ng/mL (p = 0.031) or CBDA 600 ng/mL (p = 0.017). Tumor necrosis factor (TNF)-α, monocyte chemoattractant protein (MCP-1), IL-2, and IL-18 were higher in atopic PBMC exposed to combinations of CBD, CBDA, and BCP compared to healthy post-exposure PBMC. This is the first study that tested the effect of CBD, CBDA, and BCP at different concentrations on atopic and healthy canine PBMC. The results of this study show that CBD, CBDA and BCP, at the tested concentrations, are safe for canine PBMC. However, CBD, CBDA and BCP did not show any direct anti-inflammatory effect under these experimental conditions. Further research is needed to confirm these results in a larger canine population.

Neospora caninum is a protozoan with a facultative heteroxenous life cycle, with canids as the definitive hosts and other mammals, mainly ruminants, acting as intermediate hosts. This parasite is recognized as one of the major abortifacient pathogens in cattle. Although reproductive disorders have also been reported in other domestic and wild ruminants, epidemiological data on N. caninum in ruminant species other than cattle is still limited. Here, we evaluate the seroprevalence and risk factors associated with N. caninum exposure in sheep (Ovis aries) and European mouflon (Ovis aries musimon) in southern Spain. Serum samples from 390 sheep and 387 free-ranging mouflons were tested for antibodies against N. caninum using in-house time-resolved fluorescence immunoassay based on NcGRA7 as a recombinant antigen. The individual seroprevalence was 26.2 % (95 %CI: 22.0-30.7) in sheep and 5.7 % (95 %CI: 3.8-8.5) in mouflons. At least one seropositive animal was detected in all the 26 sheep farms (100 %) and in eight of the 18 (44.4 %) hunting estates sampled. The presence of dogs on the farm (≥3) and the sheep breed (purebred) were potential risk factors associated with exposure to N. caninum in sheep. Our results indicate a high circulation of N. caninum in sheep farms in southern Spain. Control measures should be implemented to limit the exposure to this protozoan in sheep flocks. Although we have confirmed for the first time the presence of anti- N. caninum antibodies in European mouflon in Spain, and, despite the relatively low seroprevalence found, the impact of this parasite on mouflon populations should be further studied.

In this study we present the first in vitro model based on a muscle-derived cell line from a male Cuvier's Beaked whale stranded along the Italian coastline. In the myogenic cell line at T0 stage we performed the growth rate assay, the karyotyping, the CBA chromosome banding, G banding, nucleolar organizer regions and telomere analysis and immunocytochemical analysis. The cell characterization at the T2 and T4 stage included: assessment of fusion index, the ultrastructural analysis and immunocytochemical analysis. The population doubling time was determined to be ∼54 h. The cell fusion index at the T2 stage was 8.2 +/- 5 %, at the T4 stage increased at index 28 +/- 10 %. The karyotype analysis revealed a 2n = 42, XY, two pairs of nucleolus organizer regions (NORs), characteristic CBA-banding and PNA-telomeric regions by FISH-mapping. The immunocytochemical results revealed that we generated a heterogeneous population of myogenic cells. A cell population express β-actin, myosin and vimentin and a sub-population of cells was desmin-positive. In details, at the T0 stage, both cell populations were undifferentiated mononucleated myoblast. At the T2 and T4 stage, the cells were capable of fusion in elongated multinucleated myotubes, probably resulting from the fusion of the myoblasts. This Cuvier's Beaked whale cell line, represents a new opportunity to better understand the physiological features of the cetacean Ziphius cavirostris myogenic cells. The possibility of setting up culture conditions that mimic the in vivo microenvironment, strengthen the importance of in vitro models for toxicological studies investigating water pollutants effects on cells.

Mastitis is an inflammatory udder disease that causes important economic losses in the animal breeding and dairy product industries. While antibiotics have contributed to disease control, their use presents risks due to drug residues in livestock products that can potentially threaten human health. Paeonol, an active ingredient extracted from the dried root bark of paeonia suffruticosa, exhibits various pharmacological effects, including anti-inflammatory and antibacterial properties, but its role in mastitis remains unclear. In this study, we first evaluated the effects of paeonol on LPS-induced mastitis in mice and further explored the potential anti-inflammatory mechanism of paeonol on bovine mammary epithelial cells (BMECs). The results showed that paeonol alleviated LPS-induced mouse mastitis by inhibiting the infiltration of inflammatory cells and the release of inflammatory factors. Cellular assays further demonstrated that paeonol reduced apoptosis and inhibited inflammation and oxidative stress injury by mechanisms involving activation of Nrf2 signaling and inhibition of MAPK signaling in BMECs. Collectively, our results suggest that paeonol holds promise as a therapeutic agent for alleviating LPS-induced mastitis injury while providing novel insights into potential drug targets for mastitis treatment.

As adult stem cells with various advantages, Bone marrow-derived mesenchymal stem cells (BMSCs) are valuable resources for veterinary treatment and animal reproduction. Previous studies have shown that hypoxia can induce epithelial-mesenchymal transition (EMT) and improve mesenchymal characteristics of BMSCs in vitro culture. However, the mechanism by which hypoxia improves the interstitial characteristics of buffalo BMSCs (bBMSCs) remains unclear. In this study, the effects of hypoxia on the mesenchymal characteristics of bBMSCs and the expression level of lncRNA H19 were examined, and then the effects of lncRNA H19 on maintaining the mesenchymal characteristics of bBMSCs under hypoxic culture conditions (5 % oxygen) as well as its mechanism also were explored, so as to further understand the molecular mechanism of mesenchymal characteristics maintenance of bBMSCs. The results showed that hypoxic culture conditions promoted EMT of bBMSCs, with lncRNA H19 expression up-regulated. When lncRNA H19 was knocked down in hypoxia, the expression level of Vimentin was down-regulated, the expression level of E-Cadherin was up-regulated, and EMT was inhibited. Meanwhile, the genes (p-PI3K and p-AKT1) involved in PI3K/AKT signaling pathway were inhibited by lncRNA H19 Knockdown. IGF-1 (10 ng/mL), an activator of PI3K/AKT signaling pathway, was added to rescued the inhibition of PI3K/AKT signaling pathway caused by lncRNA H19 Knockdown, with the effects of lncRNA H19 on EMT related genes also partially reversed. These findings not only provide theoretical guidance to elucidate the detailed regulation mechanism of hypoxia on mesenchymal nature maintenance of bBMSCs, but also provide positive support to further establish the stable in vitro culture system of bBMSCs.

Chronic exposure to ultraviolet (UV) light can cause cutaneous damage, resulting in specific pathological changes such as actinic keratosis and dermatitis. Despite actinic dermatosis being well documented in both humans and animals, it has rarely been reported in non-human primates (NHPs). Here, we describe a case of chronic UV light exposure in cynomolgus macaque (Macaca fascicularis). An adult female was presented with a seven-month history of multifocal irregular alopecic, erythematous and hyperkeratotic plaques on the dorsum with itching. Skin biopsies were sampled for histopathological analysis. A biopsy of normal skin from the neck region of an adult macaque was used as a control. Histopathological analysis revealed epidermal changes ranging from moderate to marked hyperplasia associated with hyperkeratosis and mild stratification disorders. Fibrosis was observed in the upper dermis, and multiple areas of elastotic material deposition were confirmed in the mid and deep dermis by Van Gieson special staining. The diagnosis was "solar dermatitis with elastosis". Systemic treatment with nutritional supplements (retinol and omega-3) and firocoxib (5 mg/kg orally once a day for 3 months) did not show any improvement. Concurrently, a topical therapy with aloe gel and a sunscreen spray (SPF50) was added. The lesions did not show further clinical progression. During the period of the study, two other cynomolgus macaques from the same enclosure developed comparable clinical lesions on the dorsum. The authors describe UV-light induced skin damage in a cynomolgus macaque, which remained clinically stable.

Staphylococcus aureus is the main etiologic agent of contagious dairy cow mastitis, while non-aureus staphylococci and mammaliicocci (NASM) are the bacteria most frequently isolated from milk. Beyond their impact on animal health, NASM can harbor antimicrobial resistance (AMR) genes with potential for bidirectional transfer with S. aureus, and methicillin-resistant (MR) staphylococci (MRS) can raise significant One Health concerns. In our study, we evaluated the prevalence and characteristics of MRS in the bulk tank milk (BTM) of 88 dairy farms in the livestock-dense province of Lodi, Lombardy, northern Italy. S. aureus was isolated from 32.95 % of BTM samples, with the Ribosomal Spacer PCR (RS-PCR) genotype B being the most prevalent, identified in 37.93 % of S. aureus positive farms. All isolates carried the ica genes (icaA, icaB, icaC, icaD) indicating the potential to produce biofilm. MRS were isolated in 56.81 % of farms. According to MALDI-TOF MS analysis, the most prevalent MR species included S. epidermidis (MRSE, 35.59 %) followed by S. aureus (MRSA, 18.64 %), M. sciuri (15.25 %), S. saprophyticus (11.86 %), S. borealis (6.78 %), S. haemolyticus (5.08 %), M. fleurettii, (3.39 %), S. cohnii, and S. pettenkoferi (1.70 % each). Most MR isolates carried the mecA gene, while none carried mecC. The staphylococcal cassette chromosome mec (SCCmec) was predominantly type V in MRSA (45.45 %) and type IV in MRSE (61.90 %). Given their relevance to One Health, monitoring AMR in all staphylococci and mammaliicocci isolated from milk is essential for understanding the prevalence, characteristics, and transmission dynamics of MR gene pools within dairy herds.

Osteoarthritis (OA) is a progressive degenerative disease in older cats, and often leads to decreased quality of life (QOL). Mesenchymal stem cells (MSCs) have been used in novel therapies for inflammatory diseases. We aimed to evaluate quantitatively allogeneic adipose-derived MSC (ADSC) therapy in cats with naturally occurring OA, based on QOL assessment resources. To characterize the in vitro properties of ADSCs, we estimated ADSCs from four healthy cats with respect to morphology, differentiation potential, and immunomodulatory potential. Six cats with OA were administered a single intravenous injection of allogeneic ADSCs. Based on the feline musculoskeletal pain index (FMPI), the outcome measure was QOL. The cultured cells were adherent, exhibited a spindle shape without becoming flattened or large, and maintained doubling time until passage 5. After induction, the cells had osteogenic, adipogenic, and chondrogenic phenotypes. These cells expressed CD44 and CD90 and lacked expression of CD14 and CD45, had significantly suppressed the production of interferon -ɤ released from mitogen-stimulated lymphocytes (P < 0.05). The FMPI of all cats with OA significantly increased one month after ADSC therapy (P < 0.05). No adverse effects associated with ADSC administration were observed during follow-up in any of the cats. In conclusion, ADSC therapy with immunomodulatory potential could have beneficial effects on the QOL in cats with OA. Further research is necessary to carry out larger studies of the effectiveness of ADSC therapy.

Unesp-Botucatu Cattle Pain Scale (UCAPS) is widely used in experimental settings, however the high number of UCAPS behaviors might represent a barrier to its implementation in the farm's or hospital's routine. We aimed to identify a smaller combination of UCAPS behaviors that could be used as behavioral red flags for optimizing the acute pain diagnosis in cattle. We hypothesize that a specific set of UCAPS behaviors might be used as behavioral red flags for pain. This would represent a quick and simple pain evaluation and might optimize the acute pain assessment in large-scale systems. Data from two previous studies regarding UCAPS assessments before (pain free condition) and after (painful condition) surgical castration of 59 male cattle was used. We fitted a decision tree, resulting in a logic with two behaviors that we used as behavioral red flags. The logic adapted from the decision tree considered the painful diagnosis when the Activity was scored 2. When Activity was scored less than 2, but Locomotion was scored 1 or 2, the diagnosis was also considered positive for pain. When Activity was below 2 and Locomotion was 0, the diagnosis was considered free pain. Behavioral red flags had an area under the curve of 95.95 % for predicting UCAPS diagnosis and 94.13 % for predicting overall pain free and painful conditions. In conclusion, behaviors in the decision tree logic can work as behavioral red flags for optimizing the acute pain diagnosis in cattle, as a quick assessment in large-scale systems.

It has been shown that different production systems (conventional vs. pasture-raised pigs) and co-rearing of pigs with other livestock influence the gut microbiota composition in pigs. In this study, two independent trials were conducted to investigate the 16S fecal microbiota of the autochthonous Krškopolje pig. In each trial, three study groups were analyzed: (i) pasture-raised pigs cohabiting with small ruminants (group L1, farm 1), (ii) indoor-raised pigs without contact with other farm animals (group L2, farm 2) and fed the same commercial organic feed as the other two groups, and (iii) pasture-raised pigs cohabiting with cattle (group L3, farm 2). The pigs were sampled four times during the grower-finisher period, corresponding to the different seasons. A total of 18 and 22 pigs were included in trials 1 and 2, respectively. Alpha diversity was comparable between the study groups and the pigs of different age categories. The two predominant bacterial phyla in all three groups were Bacteroidota and Firmicutes. Significant differences in microbiota composition were found between pigs of different age categories in both trials (nonparametric MANOVA, p < 0.008). The microbiota composition of pigs from group L1 was significantly different from that of pigs from groups L2 and L3, whereas groups L2 and L3 tended to be more similar in both trials. The present results indicate that the trial and the farm of origin have a significant influence on the pig gut microbiota, and that their influence is more pronounced than that of the housing system.

There are a limited number of chemotherapeutic agents for the treatment canine visceral leishmaniasis (CVL). This study aimed to formulate and assess micellar formulation of miltefosine (MTF) to increase antileishmanial therapeutic effects in dogs naturally infected by Leishmania (L) infantum for the first time. MTF loaded pluronic F127 (PF/MTF) micelles were formulated and coated with chitosan (Cs/PF/MTF). Toxicity of compounds was evaluated. The in vitro and in vivo antileishmanial effect of Cs/PF/MTF was examined in L. infantum promastigotes and amastigotes as well as naturally infected dogs by L. infantum. In-vitro efficacy studies in promastigotes and amastigotes demonstrated that inhibitory concentration (IC50) of Cs/PF/MTF was significantly lower in comparison to miltefosine. In vivo data showed that Cs/PF/MTF is more effective against CVL compared with miltefosine alone. The result of this study indicated that intravenous injection of new formulation of Cs/PF/MTF was tolerated and treated on L. infantum naturally infected dogs with the dosages of 170 μg/kg daily for 28 days.

Classical swine fever (CSF) re-emerged in Japan in 2018, with the epidemic virus identified as genotype 2.1, which is moderately virulent and more difficult to detect and control than the highly virulent strain. Domestic pigs were administered with GPE strain live attenuated vaccine (GPE vaccine) for outbreak management. CP7_E2alf strain recombinant live vaccine (marker vaccine), approved for differentiation of infected from vaccinated animals (DIVA), was considered optional for obtaining CSF-free country certification issued by the World Organization for Animal Health. This study aimed to assess the efficacy of both vaccines in pigs through experimental challenge tests and evaluate the DIVA ability of the marker vaccine using two enzyme-linked immunosorbent assay (ELISA) antibody detection kits. Results showed that both GPE and marker vaccines were effective against the Japanese epidemic strain; however, the ability of the ELISA antibody detection kits to discriminate the marker vaccine was limited. Therefore, to achieve CSF-free certification using vaccines with DIVA functionality, alternative detection methods and enhancement of the sensitivity and specificity of ELISA kits are needed.

Microcirculation is frequently assessed using videomicroscopy in the sublingual mucosa. However, limited research has been conducted on the existence of blood flow autoregulation in this region. We conducted a study in an experimental porcine model of pharmacologically induced hypotension to evaluate the relationship between mean arterial pressure (MAP) and the microvascular flow index (MFI). We hypothesized that this relationship would be linear in the absence of autoregulation or bilinear if autoregulation is present. Seven pigs underwent blood pressure changes induced by norepinephrine (hypertension) and sevoflurane (hypotension) administration. Sublingual microcirculation was assessed using a sidestream dark field device, and videos were recorded at different MAP levels ranging from 30 to 110 mmHg. MFI was calculated using the quadrant-based method. For our first hypothesis, we constructed a linear mixed model, while a bilinear model was used for the second hypothesis. The linear model demonstrated a statistically significant association (P = 0.03) described by the equation: MFI = 2.29 + 0.004 x MAP. The bilinear model identified a statistically significant inflection point at MAP = 99 mmHg (P = 0.01) with MFI = 2.7 AU (P < 0.0001). For MAP <99 mmHg, the relationship was: MFI = 2.26 + 0.004 x MAP, and for MAP >99 mmHg, MFI = 2.7. Despite statistical significance, neither model provided a satisfactory graphical fit due to high inter- and intra-individual variability. Consequently, this study did not allow us to draw conclusions regarding the presence of blood autoregulation.

Toxoplasma gondii is one of the main pathogens causing abortion in sheep. In this study, an in house ELISA using recombinant GRA1 and BAG1 proteins as antigen was used to detect T. gondii infection in a flock of sheep suffering from abortions. Serum samples collected from sheep (n = 23) the day after abortion were initially analyzed with a commercial ELISA using recombinant P30 (SAG1) protein as antigen and then by the in house ELISA using recombinant GRA1 and BAG1 proteins as antigen. Commercial ELISA detected anti-T. gondii IgG antibodies in 20 samples whereas in house ELISA detected anti-T. gondii IgG antibodies in 19 samples. The results of the remaining three serum samples that were seronegative by commercial ELISA were also seronegative by in house ELISA. According to the results of commercial ELISA and in house ELISA, the seroprevalence of ovine toxoplasmosis was 86.9% and 82.6%, respectively. When we accept the validated commercial ELISA as a reference method, the sensitivity and specificity of in house ELISA can be considered as 95% and 100%. These preliminary results show that in house ELISA using recombinant GRA1 and BAG1 proteins as antigen can detect ovine toxoplasmosis successfully.