Spread and persistence of antimicrobial resistance genes in wastewater from human and animal sources in São Paulo, Brazil.

The spread of antimicrobial resistance (AMR) through multiple reservoirs is a global concern. Wastewater is a critical AMR dissemination source, so this study aimed to assess the persistence of resistance genetic markers in wastewater using a culture-independent approach. Raw and treated wastewater samples (n = 121) from a wastewater treatment plant (WWTP), a human hospital, a veterinary hospital, and a pig farm were monthly collected and concentrated by filtration. DNA was extracted directly from filter membranes, and PCR was used in the qualitative search of 32 antimicrobial resistance genes (ARGs). Selected genes (blaCTX-M, blaKPC, qnrB, and mcr-1) were enumerated by quantitative real-time PCR (qPCR). Twenty-six ARGs were detected in the qualitative ARGs search, while quantitative data showed a low variation of the ARG's relative abundance (RA) throughout the months, especially at the human hospital and the WWTP. At the WWTP, despite significantly reducing the absolute number of gene copies/L after each treatment stage (p < 0.05), slight increases (p > 0.05) in the RAs of genes blaCTX-M, qnrB, and mcr-1 were observed in reused water (tertiary treatment) when compared with secondary effluent. Although the increase is not statistically significant, it is worth noting that there was some level of ARGs concentration after the disinfection process. No significant absolute or relative after-treatment quantification reductions were observed for any ARGs at the veterinary hospital or the pig farm. The spread of ARGs through sewage needs to be continuously addressed, because their release into natural environments may pose potential risks of exposure to resistant bacteria and impact local ecosystems.

Evaluation of totally implantable catheters in healthy horses.

BACKGROUND: For horses requiring prolonged daily cephalic intravenous regional limb perfusion (IVRLP), the use of a totally implantable catheter (TIC) could be indicated to reduce complications associated with frequent venipuncture or external catheterization. This study aims to evaluate the implantation technique of the TIC in the cephalic vein of horses for IVRLP, describe the complications associated with the device's placement and use, and assess its viability up to 60 days after implantation. Totally implantable catheters, cut to 15 cm (n = 5) and 46 cm (n = 5) in length, were implanted into one cephalic vein in ten adult horses (n = 10). Twenty-four hours following placement, IVRLP with contrast was performed via the TIC and evaluated with radiography. Physical examinations, lameness evaluation, hematologic assessment, and the catheter patency tests were performed at scheduled intervals for the duration of catheterization (7-60 days). RESULTS: Catheters were implanted without difficulty and allowed for IVRLP 24 h post implantation. Complications resulted in removal of the catheters, with four maintained for 7 days, three in place for 15 days, and three catheters maintained for 60 days. Complications included lameness, limb swelling, catheter kinking, and venous thrombosis. CONCLUSIONS: The implantation technique of the TIC in the cephalic vein of horses is feasible and requires minimal technical effort. Although TIC allows venous access without the need for repeated venipuncture, its long-term use presents complications. For horses requiring prolonged daily cephalic IVRLP, the use of a TIC could be indicated. However, the high incidence of venous thrombosis may limit clinical application.

The Best Protocol to Treat Equine Skin Wounds by Second Intention Healing: A Scoping Review of the Literature.

Equine skin wound treatment continues to be a challenge for veterinarians. Despite being a frequent practice, it remains difficult to choose an evidence-based treatment protocol. This study aimed to comprehensively explore the literature and provide a scoping review of therapeutic strategies for equine skin wounds and identify knowledge gaps and opportunities for future research. This review was conducted using specific criteria to select literature that described methods to manage second intention wound healing. After removing duplicates and screening papers for suitability, 81 manuscripts were included for data extraction. Of these, 59 articles were experimental studies, 10 were case reports, 9 were case series, and 3 were clinical studies. The most frequent wound location was the distal limbs. Macroscopic assessment was the main tool used to evaluate treatment effectiveness. All of the case reports, case series, and clinical studies reported positive outcomes with regard to the treatment used, while only 36% of the experimental studies found significant healing improvement in treated wounds compared to control groups. It was found that there are many treatments that have exhibited controversial results, and there exists a lack of evidence for the adoption of specific treatment protocols.

Effects of intra-articular administration of hyaluronic acid or platelet-rich plasma as a complementary treatment to arthroscopy in horses with osteochondritis dissecans.

Although arthroscopy is the treatment of choice for horses with osteochondritis dissecans (OCD), it is not yet known whether intra-articular therapies in the postoperative period can bring any benefit to the recovery of these animals. This study evaluated the effects of the intra-articular application of platelet-rich plasma (PRP), hyaluronic acid (HA) or lactated Ringer's solution (LR) in horses with OCD undergoing arthroscopy. Eighteen male and female Brazilian Sport horses aged between 2 and 6 years were evaluated. All animals presented OCD fragments in the middle crest of the tibia. Ten days after surgery, animals were randomly distributed into three groups and received intra-articular application of PRP (n = 6), HA (n = 6), or LR (control group, n = 6). Clinical, radiographic, ultrasound and synovial fluid evaluations were performed on the day of surgery and after 10, 30 and 60 days. An increase in the thickness of the joint capsule was observed 30 days after surgery in the three groups evaluated. In the control group, there was significant improvement in the flexion test 30 and 60 days after surgery, and in the PRP group, there was worsening of this parameter in the same evaluations. In the control group, there was a reduction in the degree of synovial effusion, and in the PRP and HA groups, there was increased effusion. There was a significant increase in the number of leukocytes in the HA group. Intra-articular use of PRP or HA ten days after arthroscopy did not promote positive effects on the recovery of horses with OCD.

Functional infrared thermography imaging can be used to assess the effectiveness of Maxicam Gel® in pre-emptively treating transient synovitis and lameness in horses.

Introduction: Diagnosing and treating lameness in horses is essential to improving their welfare. In equine orthopedic practice, infrared thermography (IRT) can indirectly detect soreness. Non-steroidal anti-inflammatory drugs can treat painful and inflammatory processes in horses. Using IRT, the efficacy of meloxicam (Maxicam Gel®) was evaluated in pre-treating transient synovitis in horses induced by a middle carpal joint injection of lipopolysaccharides (LPS) from E. coli 055:B5 at a dose of 10 endotoxin units. Methods: In a cross-over design, six healthy horses were randomly assigned to receive either 0.6 mg/kg of oral Maxicam Gel® (MAXVO) or a mock administration (control group, C) following a two-week washout period. IRT of the middle carpal joint, visual lameness assessment and joint circumference were recorded over time. Clinical and hematological evaluations were performed. Synovial fluid aspirates were analyzed for total nucleated cell count, total protein, and prostaglandin E2. A mixed effects analysis of variance was performed for repeated measures over time, followed by Tukey's test. A multinomial logistic regression was conducted to determine whether there is a relationship between a thermography temperature change and the lameness score. Results: There were no changes in joint circumference. The MAXVO group showed a lower rectal temperature 4 h after synovitis induction. The C group presented an increase in neutrophils and a decrease in total hemoglobin and hematocrit 8 h after induction. No changes were observed in the synovial fluid between groups. The horses that received meloxicam did not show clinically significant lameness at any time, while the C group showed an increase in lameness 2, 4, and 8 h after synovitis induction. Discussion: IRT indicated that the skin surface temperature of the middle carpal joint was lower in horses who received meloxicam, suggesting a reduction in the inflammatory process induced by LPS. It was observed that the maximum temperature peaks in the dorsopalmar and lateropalmar positions can be utilized to predict the severity of lameness, particularly when the temperature rises above 34°C. Horses pre-treated with meloxicam showed either reduced or no indication of mild to moderate pain and presented a lowehr thermographic temperature, which indicates the effectiveness of Maxicam Gel® as an anti-inflammatory.

Cost of transport, but not gluteus medius and red blood cells monocarboxylate-transporters density differentiated Brazilian Sport Horses at two performance levels.

Cost of transport (COT) and monocarboxylate transporters (MCTs) could affect the ability to perform fast actions during a jumping discipline. This study aimed to compare the COT and evaluate the MCT1, MCT4, and their auxiliary protein CD147 content in the gluteus medius and RBCs of Brazilian sport horses (BH), a breed developed for jumping competitions, with low-level (LL) or intermediate-level (IL) jumping capacities. The physiological difference between the horses was assessed by an incremental jump test (IJT), in which the cost of lactate (COTLAC) and heart rate (COTHR) of running were determined for each animal by the ratio between each variable and the running speed. Western blotting was performed on muscle and RBC membranes to quantify MCT1, MCT4, and CD147. IL showed lower COTLAC and COTHR than LL at all jumping heights. The amount of MCT1, MCT4, and CD147 found in muscle and RBCs were not dependent on performance level. Muscle MCT4 and MCT1 were correlated positively with CD147. We conclude that the relatively small differences between performances did not relevantly influence MCT expression in BH. While MCT analyses are inaccessible for most trainers and veterinarians, the cost of transport measurements is a feasible and sensitive tool to distinguish intermediate and low-level jumping horses.

Intravenous Regional Limb Perfusion in Standing and Recumbent Horses: A Comparative Radiographic Study.

Although pharmacokinetic studies of drugs administered by intravenous regional limb perfusion (IRLP) to treat equine orthopedic infections suggest efficient drug distribution in the limbs, it remains unclear whether drug perfusion is affected by the position of the horse during the procedure. This study compared the perfusion of a radiopaque contrast into tissues of the extremities of horses maintained in standing and recumbent positions during an IRLP. Radiopaque contrast was administered through IRLP into the cephalic vein of 10 healthy adult horses under general anesthesia and right lateral recumbency (RG) or under sedation and standing (SG). The same animals were used in both groups, respecting a two-week washout period. Sequential radiographic images were performed immediately at the beginning of contrast administration (T0) and after 10, 20, 30, 40, and 50 minutes. Tourniquets were removed after 30 minutes. The time required for the contrast to reach the hooves was compared between groups. Contrast reached the hooves faster in SG (114 ± 15 seconds) compared with RG (236 ± 29 seconds) (P < 0.5). SG showed more uniform perfusion of the limb vessels, whereas RG showed more deposition of the contrast in the lateral digital vein, with smaller amounts reaching the hooves. From T10 onward, soft tissue radiopacity increased, albeit more markedly in standing than in recumbent animals, remaining until T50. Contrast radiography evidenced that IRLP performed in standing position leads to a quicker and more uniform perfusion of the vasculature and a more noticeable diffusion to the tissues than in recumbent horses.

Differential Expression of Monocarboxylate Transporter 1 and Ancillary Protein CD147 in Red Blood Cells of Show Jumping Horses.

We compare the expression levels of the lactate transporter complex consisting of the lactate transporter, monocarboxylate transporter 1 (MCT1), and its ancillary protein, cluster of differentiation 147 (CD147), in the membranes of red blood cells (RBCs) from two breeds of jumping horses and associate the expression levels of these proteins with their jumping ability. The expression levels of MCT1 and CD147 proteins on the membranes of RBCs collected from 30 show jumping horses of two different breeds were quantified: the Brazilian Sport Horses (n = 17) and the European Warmbloods (n = 13). The levels of MCT1 and CD147 in the RBC membranes were measured by Western blot using horse-specific antibodies. Statistical analyses included unpaired Student t-test and Chi-squared test. According to the expression levels of MCT1 and CD147 proteins, 88% of the Brazilian Sport Horses were categorized as high lactate transporters and the remaining 12% as low lactate transporters. The opposite was found for the European Warmbloods, where most animals (77%) were classified as low lactate transporters and the remaining animals (23%) were classified as high lactate transporters. Brazilian Sport Horses express statistically significantly higher levels of CD147 and MCT1 than European Warmbloods. The classification of horses considering the expression of proteins involved in the ability to transport lactate through the complex MCT1-CD147 seems to be breed dependent, with horses that are able to jump higher obstacles showing lower expression of the MCT1-CD147 complex in their RBCs.

Occurrence of gastric ulcers in horses exercised on a treadmill / Ocorrência de úlceras gástricas em cavalos exercitados em esteira rolante

ABSTRACT: The most frequently causes of gastric ulcers in horses are fasting, feeding type and management, as well as administration of certain medications. More recently, exercise has also been identified as an important factor in the development of gastric lesions in athlete horses. The purpose of the present study was to assess whether treadmill exercise predisposes the horses to the occurrence of gastric ulcers. Five Arabian horses underwent a twelve-week training period on a treadmill and a gastroscopic evaluation before and after the training period. All animals showed gastric lesions after the training period. Based on these results, it is concluded that Arabian horses undergoing exercise on a treadmill may develop gastric lesions varying in number and severity.

RESUMO: As causas mais frequentes de úlceras gástricas em cavalos são jejum, tipo de alimentação e manejo, bem como a administração de alguns medicamentos. Mais recentemente, o exercício também foi identificado como um fator importante no desenvolvimento das lesões gástricas em cavalos atletas. O objetivo do presente estudo foi avaliar se o exercício em esteira rolante predispõe os cavalos à ocorrência de úlceras gástricas. Cinco cavalos árabes foram submetidos a um período de doze semanas de treinamento em esteira rolante e a avaliação gastroscópica antes e após o período de treinamento. Todos os animais apresentaram lesões gástricas após o treinamento. Com base nesses resultados, conclui-se que cavalos árabes submetidos a exercício em esteira rolante podem desenvolver lesões gástricas em número e gravidade variados.

Culture of equine bone marrow mononuclear fraction and adipose tissue-derived stromal vascular fraction cells in different media / Cultivo de células da fração mononuclear da medulla óssea e da fração vascular stromal do tecido adipose de equinos em diferentes meios

The objective of this study was to evaluate the culture of equine bone marrow mononuclear fraction and adipose tissue - derived stromal vascular fraction cells in two different cell culture media. Five adult horses were submitted to bone marrow aspiration from the sternum, and then from the adipose tissue of the gluteal region near the base of the tail. Mononuclear fraction and stromal vascular fraction were isolated from the samples and cultivated in DMEM medium supplemented with 10% fetal bovine serum or in AIM-V medium. The cultures were observed once a week with an inverted microscope, to perform a qualitative analysis of the morphology of the cells as well as the general appearance of the cell culture. Colony-forming units (CFU) were counted on days 5, 15 and 25 of cell culture. During the first week of culture, differences were observed between the samples from the same source maintained in different culture media. The number of colonies was significantly higher in samples of bone marrow in relation to samples of adipose tissue.

O objetivo deste estudo foi avaliar o cultivo de células da fração mononuclear da medula óssea e da fração vascular estromal do tecido adiposo de equinos em dois diferentes meios. Cinco cavalos foram submetidos à aspiração da medula óssea do esterno e à coleta de tecido adiposo da região glútea, próxima à inserção da cauda. A fração mononuclear e a fração vascular estromal foram obtidas das amostras e cultivadas em meio DMEM suplementado com soro fetal bovino a 10% ou em meio AIM-V. As culturas foram observadas uma vez por semana com um microscópio de luz invertida, com o intuito de se realizar uma análise qualitativa das características morfológicas das células, bem como do aspecto geral do cultivo celular. As unidades formadoras de colônia (CFU) foram contadas nos dias 5, 15 e 25 do cultivo celular. Durante a primeira semana, foram observadas diferenças entre amostras obtidas de mesma origem mantidas em diferentes meios. O número de colônias foi significativamente maior nas amostras de medula óssea em relação às amostras de tecido adiposo.

Serial superficial digital flexor tendon biopsies for diagnosing and monitoring collagenase-induced tendonitis in horses / Biópsias seriadas do tendão flexor digital superficial para o diagnóstico e monitoração da tendinite induzida por colagenase em equinos

The purpose of this investigation was to demonstrate the feasibility of a biopsy technique by performing serial evaluations of tissue samples of the forelimb superficial digital flexor tendon (SDFT) in healthy horses and in horses subjected to superficial digital flexor tendonitis induction. Eight adult horses were evaluated in two different phases (P), control (P1) and tendonitis-induced (P2). At P1, the horses were subjected to five SDFT biopsies of the left forelimb, with 24 hours (h) of interval. Clinical and ultrasonographic (US) examinations were performed immediately before the tendonitis induction, 24 and 48 h after the procedure. The biopsied tendon tissues were analyzed through histology. P2 evaluations were carried out three months later, when the same horses were subjected to tendonitis induction by injection of bacterial collagenase into the right forelimb SDFT. P2 clinical and US evaluations, and SDFT biopsies were performed before, and after injury induction at the following time intervals: after 24, 48, 72 and 96 h, and after 15, 30, 60, 90, 120 and 150 days. The biopsy technique has proven to be easy and quick to perform and yielded good tendon samples for histological evaluation. At P1 the horses did not show signs of localised inflammation, pain or lameness, neither SDFT US alterations after biopsies, showing that the biopsy procedure per se did not risk tendon integrity. Therefore, this procedure is feasible for routine tendon histological evaluations. The P2 findings demonstrate a relation between the US and histology evaluations concerning induced tendonitis evolution. However, the clinical signs of tendonitis poorly reflected the microscopic tissue condition, indicating that clinical presentation is not a reliable parameter for monitoring injury development. The presented method of biopsying SDFT tissue in horses enables the serial collection of material for histological analysis causing no clinical signs and tendon damage seen by US images. Therefore, this technique allows tendonitis to be monitored and can be considered an excellent tool in protocols for evaluating SDFT injury.

Objetivou-se demonstrar a viabilidade de uma técnica de biópsia na realização de avaliações sequenciais de amostras de tecido do tendão flexor digital superficial (TFDS) do membro torácico, em equinos hígidos e em equinos submetidos à indução de tendinite do TFDS. Oito equinos adultos foram avaliados em duas fases (F) diferentes, controle (F1) e apresentando tendinite induzida (F2). Na F1, os equinos foram submetidos a cinco biópsias do TFDS do membro torácico esquerdo, com 24 horas (h) de intervalo entre as mesmas. Avaliações clínicas e ultrassonográficas (US) foram realizadas imediatamente antes e, 24 e 48 h após cada procedimento. O tecido tendíneo coletado por biópsia foi analisado histologicamente. A F2 ocorreu três meses depois, quando os mesmos equinos foram submetidos à indução de tendinite do TFDS do membro torácico direito por injeção intratendínea de colagenase. Na P2, Avaliações clínicas e US foram realizadas antes da indução da tendinite e após a mesma nos seguintes momentos: 24, 48, 72 e 96 h depois e, 15, 30, 60, 90, 120 e 150 dias depois. A técnica de biópsia se demonstrou ser de fácil e rápida realização, fornecendo fragmentos de tecido tendíneo adequados para a realização de histologia. Na F1, os equinos não demonstraram sinais de inflamação local, dor ou claudicação, bem como não apresentaram alterações na avaliação US do TFDS após as biópsias, demonstrando que o dano físico ao tendão provocado pela biópsia não compromete sua integridade. Assim, o procedimento pode ser utilizado rotineiramente para avaliações histológicas do tecido tendíneo. Na F2, observou-se uma relação entre os achados US e histológicos no que se refere à evolução da tendinite induzida. Entretanto, observou-se que os sinais clínicos da tendinite não acompanham a condição microscópica do tecido, o que indica que a apresentação clínica não pode ser utilizada como parâmetro para a monitoração da evolução da enfermidade. A técnica de biópsia apresentada para colheita de tecido do TFDS em equinos, permite a obtenção de tecido para avaliação histológica seriada sem provocar sinais clínicos e alterações ultrassonográficas que indiquem dano tecidual. Assim, a técnica permite a monitoração da tendinite e pode ser considerada uma excelente ferramenta na avaliação de injúrias do TFDS.

Viabilidade celular da fração mononuclear da medula óssea e fração vascular estromal do tecido adiposo de equinos após o processo de congelamento e descongelamento / Viability of equine bone marrow mononuclear fraction and adipose tissue-derived stromal vascular fraction after freezing and thawing process

Cinco cavalos adultos foram submetidos à coleta de medula óssea do esterno e de tecido adiposo da região glútea. As amostras foram processadas para obtenção da fração mononuclear da medula óssea e fração vascular estromal do tecido adiposo, o número de células obtidas e a viabilidade celular foram determinados. Em seguida, realizou-se o congelamento das amostras em solução contendo 20% de soro fetal bovino e 10% de dimetilsulfóxido. Depois de um mês, realizou-se o descongelamento das amostras e a viabilidade celular foi novamente mensurada. Os resultados revelaram que as técnicas utilizadas tanto para coleta de medula óssea quanto de tecido adiposo em equinos são simples, rápidas e seguras. As metodologias adotadas para o processamento das amostras foram eficientes, obtendo-se aproximadamente 95% de viabilidade celular. Após o descongelamento, a viabilidade média das amostras de células mononucleares da medula óssea foi de 86% e da fração vascular estromal do tecido adiposo de 64%. Frente à importância da terapia celular na clínica médica de equinos, concluiu-se que é necessária a realização de mais estudos, visando padronizar uma técnica de criopreservação que mantenha a integridade das células da fração mononuclear da medula óssea e da fração vascular estromal do tecido adiposo de equinos.(AU)

In five adult horses, bone marrow was aspirated from the sternum and adipose tissue extracted from the gluteal region. The samples were processed to obtain the mononuclear fraction of bone marrow and stromal vascular fraction of adipose tissue, and the number of cells obtained and cell viability were determined. Next, the cell samples were frozen in medium containing 20% fetal bovine serum and 10% dimethylsulfoxide. After one month, the cells were thawed and cell viability was again determined. The results revealed that the techniques for collecting both bone marrow and adipose tissue in horses are simple, rapid and safe. The methods used for processing the samples were efficient, yielding about 95% cell viability. After freezing, mean viability of the mononuclear cells of bone marrow was 86%, and 64% for the stromal vascular cells of adipose tissue. In view of the importance of cell therapy in equine clinical medicine, it is concluded that further studies are needed for the standardization of a cryopreservation technique to maintain the integrity for the mononuclear fraction of bone marrow and the stromal vascular fraction of adipose tissue in horses.(AU)

Anestesia por isofluorano em eqüinos submetidos à infusão contínua de medetomidina ou xilazina / Isoflurane anesthesia in horses during medetomidine or xilazine continuous infusion

Avaliaram-se oito eqüinos sob anestesia geral inalatória com isofluorano (1CAM) e infusão contínua de xilazina (0,35mg kg-1h-1) ou medetomidina (3,5µg kg-1h-1), em relação à freqüência cardíaca, ritmo cardíaco, freqüência respiratória, pressão arterial, hemogasometria arterial e temperatura, nos tempos T0 (imediatamente antes do início da infusão contínua) e T10 ao T60 (intervalos de 10 minutos, após início da infusão contínua). Houve redução da freqüência cardíaca e da temperatura e elevação da pressão arterial média. A paCO2 (no GM) elevou-se e a paO2 mostrou-se maior no GM que no GX. Conclui-se que a infusão contínua de doses equipotentes de xilazina e medetomidina, durante anestesia geral inalatória, com isofluorano, em eqüinos, promove alterações cardiocirculatórias, respiratórias, térmicas e hemogasométricas discretas e equivalentes.

Eight horses under inhalant general anesthesia with isoflurane (1MAC) and continuous infusion of xylazine (0.35mg kg-1h-1) or medetomidine (3.5µg kg-1h-1) were evaluated for heart rate and rhythm, respiratory rate, arterial blood pressure, arterial blood gas analysis and temperature immediately before the beginning of the continuous infusion (T0) and in intervals of 10 minutes after the beginning of the continuous infusion (T10 to T60). Heart rate and temperature decreased and mean arterial pressure increased. PaCO2 (in GM) increased and GM showed a higher paO2 than GX. We conclude that equipotent doses of continuous infusion of medetomidine and xylazine during inhalant general anesthesia with isoflurane in horses promote slight and equivalent cardiocirculatory, respiratory, thermic and arterial blood gases changes.