Protective effects of genistein on the production performance and lipid metabolism disorders in laying hens with fatty liver hemorrhagic syndrome by activation of the GPER-AMPK signaling pathways.

The aim of this study was to evaluate the beneficial effects and potential mechanisms of genistein (GEN) on production performance impairments and lipid metabolism disorders in laying hens fed a high-energy and low-protein (HELP) diet. A total of 120 Hy-line Brown laying hens were fed with the standard diet and HELP diet supplemented with 0, 50, 100, and 200 mg/kg GEN for 80 d. The results showed that the declines in laying rate (P < 0.01), average egg weight (P < 0.01), and egg yield (P < 0.01), and the increase of the ratio of feed to egg (P < 0.01) induced by HELP diet were markedly improved by 100 and 200 mg/kg of GEN treatment in laying hens (P < 0.05). Moreover, the hepatic steatosis and increases of lipid contents (P < 0.01) in serum and liver caused by HELP diet were significantly alleviated by treatment with 100 and 200 mg/kg of GEN in laying hens (P < 0.05). The liver index and abdominal fat index of laying hens in the HELP group were higher than subjects in the control group (P < 0.01), which were evidently attenuated by dietary 50 to 200 mg/kg of GEN supplementation (P < 0.05). Dietary 100 and 200 mg/kg of GEN supplementation significantly reduced the upregulations of genes related to fatty acid transport and synthesis (P < 0.01) but enhanced the downregulations of genes associated with fatty acid oxidation (P < 0.01) caused by HELP in the liver of laying hens (P < 0.05). Importantly, 100 and 200 mg/kg of GEN supplementation markedly increased G protein-coupled estrogen receptor (GPER) mRNA and protein expression levels and activated the AMP-activated protein kinase (AMPK) signaling pathway in the liver of laying hens fed a HELP diet (P < 0.05). These data indicated that the protective effects of GEN against the decline of production performance and lipid metabolism disorders caused by HELP diet in laying hens may be related to the activation of the GPER-AMPK signaling pathways. These data not only provide compelling evidence for the protective effect of GEN against fatty liver hemorrhagic syndrome in laying hens but also provide the theoretical basis for GEN as an additive to alleviate metabolic disorders in poultry.

Fatty liver hemorrhagic syndrome (FLHS) is a nutritional and metabolic disease that seriously threatens the health and performance of laying hens, which is characterized by hepatic steatosis and lipid metabolism disorders. As an isoflavone phytoestrogen, genistein (GEN) exerts many beneficial functions, including alleviating lipid metabolism disorders and anti-inflammatory properties. However, further research is needed on the protective effect and potential mechanism of GEN on the FLHS in laying hens. Here, we found that GEN treatment improved liver injury and decline of production performance in laying hens with FLHS. Moreover, GEN treatment alleviated hepatic steatosis and lipid metabolism disorders through reducing the expression levels of mRNA related to fatty acid transport and synthesis and enhancing the mRNA expression levels of factors associated with fatty acid oxidation in FLHS layers, which may be achieved by activation of the G protein-coupled estrogen receptor­adenosine 5'-monophosphate (AMP)-activated protein kinase signaling pathways. These data not only provide compelling evidence for the protective effects and mechanisms of GEN against FLHS in laying hens but also provide the theoretical basis for GEN to alleviate other metabolic disorders in poultry.

Hirudotherapy (medicinal leeches) for treatment of upper airway obstruction in a dog.

OBJECTIVE: To describe upper airway obstruction (UAO) in a dog treated with medicinal leeches (hirudotherapy) as an ancillary therapy to hasten recovery. CASE SUMMARY: A 10-month-old neutered female Mastiff presented for acute respiratory distress. On admission, the dog was tachycardic, cyanotic, and orthopneic; stridor was audible. A 10-cm soft tissue swelling in the right ventral cervical region and bruising around the rostral mandible were noted. At the time of endotracheal intubation, the trachea was deviated to the right as a consequence of severe soft tissue swelling that was contiguous with the sublingual hematoma and cervical region, causing loss of visualization of the arytenoids. A computed tomography with contrast scan of the head, neck, and thorax was performed, showing severe soft tissue swelling of the tongue, obliteration of the common pharyngeal/laryngeal regions from suspected hemorrhage, and rightward displacement of pharynx, larynx, and proximal trachea. Marked diffuse bronchial/bronchiolar thickening associated with bronchiolectasis and diffuse opacification of the pulmonary parenchyma with regions of consolidation were noted. The dog was minimally hypercoagulable on thromboelastography. The imaging results together with results of bronchoalveolar lavage cytology supported a comorbidity of eosinophilic bronchopneumopathy. Intubation was maintained with infusions of propofol and fentanyl, with minimal changes in oropharyngeal swelling within the first 18 hours of treatment. Medicinal leeches were then applied to the sublingual and cervical regions. There was continued slow bleeding from the sites of leech detachment, and the dog was able to be extubated at 44 hours, followed by hospital discharge. NEW/UNIQUE INFORMATION PROVIDED: Leeches are utilized in human medicine for treatment of UAO. Although UAO from hemorrhage has been described in dogs, this is the first report of medicinal leeches (Hirudo verbana) as complementary treatment for sublingual hematoma that contributed to UAO.

Preparation of the peroxisome proliferator-activated receptor α polyclonal antibody: Its application in fatty liver hemorrhagic syndrome.

Peroxisome proliferator-activated receptor α (PPARα) play a key role in the regulation of metabolic homeostasis, inflammation, cellular growth, and differentiation. To further explore the potential role of PPARα in the energy homeostasis of fatty liver hemorrhagic syndrome (FLHS), we reported the prokaryotic expression and purification of chicken PPARα subunit protein, and successfully prepared a polyclonal antibody against PPARα recombinant protein. The 987 bp PPARα subunit genes were cloned into the pEASY-T3 clone vector. Then the plasmid PCR products encoding 329 amino acids were ligated to pEASY-Blunt E2 vector and transformed into BL21 to induce expression. The recombinant PPARα subunit protein, containing His-tag, was purified by affinity column chromatography using Ni-NTA affinity column. Rabbit antiserum was generated by using the concentration of recombinant PPARα subunit protein as the antigen. The results of western blotting showed that the antiserum can specifically recognize chicken endogenous PPARα protein. Immunohistochemistry and immunofluorescence showed that the PPARα mainly existed in the nucleus of hepatocytes, renal epithelial cells and hypothalamic endocrine nerve cells. More importantly, western blotting and real-time quantitative PCR indicated that FLHS significantly decreased the expression of PPARα.

Preoperative autologous blood donation and transfusion in dogs undergoing elective surgical oncology procedures with high risk of hemorrhage.

OBJECTIVE: To describe preoperative autologous blood donation (PABD) and transfusion in dogs undergoing elective surgical oncology procedures with a high risk of intraoperative hemorrhage. STUDY DESIGN: Prospective study. ANIMALS: Twelve dogs. METHODS: Dogs undergoing surgical oncology procedures associated with a high risk of hemorrhage were enrolled. Blood was collected a minimum of 6 days before surgery and separated into fresh frozen plasma (FFP) and packed red blood cells (pRBC). Dogs received FFP at the start of surgery and pRBC intraoperatively when hemorrhage ensued. The mean packed cell volume/total solids (PCV/TS) were calculated on the day of PABD preoperatively, immediately postoperatively, and 24 hours after transfusion. The dogs were monitored for transfusion-related adverse reactions, including hyperthermia, hypotension, tachycardia, bradycardia, pale mucous membranes, prolonged capillary refill time, or tachypnea/dyspnea. RESULTS: Dogs enrolled in the study underwent mandibulectomy, maxillectomy, chest wall resection, and liver lobectomy. Ten of the 12 dogs that underwent PABD received autologous transfusion at first signs of hemorrhage intraoperatively. Iatrogenic anemia was noted in two dogs (PCV 30% and 31%). The mean PCV/TS levels on the day of blood collection, preoperatively, immediately postoperatively (after transfusion), and 24 hours posttransfusion were 45.1%/7.1 g/dL, 42.2%/6.73 g/dL, 33.2%/5.42 g/dL, and 36.5%/5.65 g/dL, respectively. No dog developed transfusion-related complications. CONCLUSION: Preoperative autologous blood donation was well tolerated and led to uneventful autologous transfusion in 10 of 12 dogs. CLINICAL SIGNIFICANCE: Preoperative autologous blood donation and autologous transfusion are feasible for dogs undergoing elective surgical procedures with a high risk of hemorrhage.

Autotransfusão imediata em cão - relato de caso / Instant autotransfusion in dog - case report

As hemorragias podem levar à morte, em caso de não interrupção e recuperação da volemia. A substituição do sangue pode ser realizada por transfusão homóloga ou autóloga. Na transfusão homóloga, o sangue é obtido de um doador, na autotransfusão ou transfusão autóloga, o sangue é coletado do próprio paciente. Objetivou-se relatar a autotransfusão em um cão como um método simples, rápido e barato, e discutir esse procedimento como uma alternativa emergencial para clínicas e hospitais que não possuem bancos de sangue ou animais doadores. Foi atendido um cão Rottweiler fêmea, 42,8kg, 10 anos de idade, com queixa principal de tumor mamário e para realização de castração. Após o procedimento, a paciente apresentou hemoperitônio, sendo encaminhada para novo procedimento cirúrgico, durante o qual se observou grande quantidade de sangue livre na cavidade abdominal. Foi realizada a autotransfusão de emergência, pela técnica das duas seringas, devido à indi1111sponibilidade de sangue ou animal doador no momento do procedimento cirúrgico emergencial, demonstrando ser uma opção eficiente, econômica, de fácil acesso e segura, por ser uma transfusão normotérmica, apresentar diminuição do risco de sobrecarga circulatória e por ser o sangue compatível, devendo ser mais explorada, pois existem poucos dados descritos na literatura.(AU)

Hemorrhages can lead to death in case of non-interruption and recovery of blood volume. Blood replacement may be performed by homologous or autologous transfusion. In homologous transfusion, blood is obtained from a donor, in autotransfusion or autologous transfusion, blood is collected from the patient himself. We aimed to report autotransfusion in a dog as a simple, quick, and inexpensive method, and discuss it as an emergency alternative for clinics and hospitals that do not have blood banks or animal donors. A female Rottweiler dog, 42.8kg, 10 years of age, was treated with a primary complaint of mammary tumor and for castration. After the procedure the patient presented hemoperitoneum, being referred to a new surgical procedure, where a large amount of free blood was observed in the abdominal cavity. The emergency autotransfusion was performed by the two syringes technique, due to the unavailability of blood or donor animal at the time of the emergency surgical procedure, proving to be an efficient, economical, easily accessible and safe option because it is a normothermic transfusion, risk of circulatory overload and blood is compatible. It should be more exploited because there are few data described in the literature.(AU)

Successful management of catastrophic peripheral vascular hemorrhage using massive autotransfusion and damage control surgery in a dog.

OBJECTIVE: To describe a case of massive transfusion using unwashed, non-anticoagulated, nonsterile autologous blood in a dog with catastrophic hemorrhage from a peripheral vessel during orthopedic surgery. A damage control surgical strategy was also employed. CASE SUMMARY: A 6-year-old, 48 kg neutered male Labrador Retriever experienced massive hemorrhage after transection of a large blood vessel while undergoing femoral head and neck osteotomy. Blood was collected from clean, but not sterile, suction canisters and clots were skimmed off. The blood was then transfused back to the dog using a standard in-line blood filter. Approximately 58% of the dog's blood volume was autotransfused in less than 2 hours, thereby meeting the criteria for massive transfusion. Surgery was aborted after hemostasis was achieved by ligation of the vessel and packing of the surgical site. Two units of fresh frozen plasma were administered postoperatively due to the development of a coagulopathy. Hemoglobinuria developed but resolved within 18 hours. Three days later, completion of the surgical procedure was performed without incident. The dog was discharged 4 days after the initial surgery. Marked swelling of the affected limb developed, but resolved after the sixth day. No other significant complications developed. NEW OR UNIQUE INFORMATION PROVIDED: In this case report, the authors describe the successful management of catastrophic hemorrhage with autotransfusion performed in the absence of sterile collection, cell washing, or anticoagulation. Although not ideal, autotransfusion under these conditions can be lifesaving in situations of massive hemorrhage. This case also highlighted the employment of a damage control surgical strategy.

Improving effect of dietary soybean phospholipids supplement on hepatic and serum indexes relevant to fatty liver hemorrhagic syndrome in laying hens.

In order to investigate the effect of dietary soybean phospholipid supplement on hepatic and serum indexes relevant to fatty liver hemorrhagic syndrome (FLHS) in layers, 135 300-day-old Hyline Brown layers were randomly divided into three groups (control, pathology and prevention), and each group had 45 layers with three replicates. Birds in the three groups were respectively fed the control diet, high-energy low-protein diet and high-energy high-protein diet affixed with 3% soybean phospholipid instead of maize. Results showed in the 30th day, birds' livers in the pathology group became yellowish, enlarged in size and had hemorrhagic spots, while the prevention and control groups' layers did not have such pathological changes. Contents of triglyceride, total cholesterol, low-density lipoprotein - cholesterol, non-esterified fatty acid and malondialdehyde in serum or liver homogenate in prevention and control groups were remarkably lower than those in the pathology group (P < 0.05 or P < 0.01), as with the activities of glutamic oxalacetic transaminase and glutamic-pyruvic transaminase (P < 0.01); high-density lipoprotein - cholesterol value was strikingly higher than that of the pathology group (P < 0.01). It is suggested dietary soybean phospholipids supplement may effectively improve hepatic and blood indexes relevant to FLHS, which provides a new point for preventing FLHS occurrence rate in laying flocks and treating human non-alcohol fatty liver disease.

Autologous blood transfusion in dogs with thoracic or abdominal hemorrhage: 25 cases (2007-2012).

OBJECTIVE: To describe the use and outcome following autologous blood transfusion (ABT) in dogs. DESIGN: Retrospective study (January 2007-July 2012). SETTING: Private veterinary referral center. ANIMALS: Twenty-five dogs that underwent ABT secondary to thoracic or abdominal hemorrhage. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The hospital transaction database was searched using the keyword "autotransfusion" from January 2007 to July 2012. Data collected included signalment, body weight, etiology of hemorrhage, source and method of collection, volumes and method of ABT administration, use of anticoagulant, reported complications, and outcome. Twenty-five dogs were included for a total of 27 ABTs. Causes of hemorrhage included vascular trauma (14/25 dogs, 56%), ruptured tumor (8/25, 32%), and coagulopathy attributed to brodifacoum toxicosis (3/25, 12%). Autologous blood was collected from the abdominal (19/25, 76%), thoracic (5/25, 20%), or abdominal and thoracic cavities (1/25, 4%). Anticoagulant was added to the ABT blood in 13 of 25 (52%) cases. A median ABT volume of 29.3 mL/kg (range 2.9-406.9 mL/kg) was infused through either a 210 µm blood administration filter (21/27, 78%) or an 18 µm hemonate filter (6/27, 22%). Reported complications that may have been associated with ABT included hypocalcemia (4/17, 24%), hemolyzed serum (5/19, 26%), and prolonged coagulation times (4/5, 80%). These complications were considered of minimal clinical significance. Additional blood products were administered in 17 of 25 (68%) dogs. Seventeen (68%) dogs survived to discharge. Cause of death in the remaining cases was euthanasia or cardiac arrest secondary to uncontrollable hemorrhage. CONCLUSIONS: ABT is an adjunct to volume replacement in dogs with thoracic or abdominal hemorrhage secondary to vascular trauma, ruptured tumor, or anticoagulant rodenticide toxicosis. ABT may be used as bridge to definitive hemorrhage control, particularly when other blood products are not available or affordable. Complications may include hypocalcemia, prolonged coagulation times, and hemolysis.

Clinical findings and treatment in 63 cows with haemorrhagic bowel syndrome.

The clinical, haematological and biochemical findings, treatment and outcome of 63 cows with haemorrhagic bowel syndrome are described. The general condition and demeanor were moderately to severely abnormal in all the cows. Signs of colic occurred in 27 cows, decreased rectal temperature in 46 and tachycardia in 44. With the exception of one cow, intestinal motility was decreased or absent. Transrectal palpation revealed dilatation of the rumen in 47 cows and dilatation of the small intestine in 18. Faecal output was markedly reduced or absent, and the faeces were dark brown to black and contained blood. Nine cows were euthanized immediately after physical examination. Conservative medical therapy was instituted in two cows; however, both were euthanized a few days later because of deterioration in condition. Exploratory right flank laparotomy was carried out in 52 cows. Of these, 22 were euthanized intraoperatively because of severe lesions. In 27 cows, intestinal massage to reduce the size of blood clots was carried out; 11 were euthanized several days postoperatively because of deterioration in condition. In three other cows, intestinal resection was carried out and all survived. Of the 63 cows, 19 (30.2%) survived and were healthy at the time of discharge from the clinic.

Effect of dietary omega-3 and omega-6 fatty acids on clotting activities of Factor V, VII and X in fatty liver haemorrhagic syndrome-susceptible laying hens.

1. The relationship between concentrations of omega-3 and omega-6 fatty acids in plasma and Factor V, VII and X clotting activities was determined using a crossover feeding trial with diets supplemented with either soy oil or flax oil. 2. Laying hens on the soy diet, which is high in omega-6 fatty acids, had substantially higher clotting activity for all three factors compared to laying hens on the flax diet that was high in omega-3 fatty acids. 3. Positive associations were seen between liver haemorrhage score and the percentage of liver weight and between the percentage of liver weight and the severity of haemorrhagic and fatty changes seen on histology. 4. These results support the hypothesis that concentrations of omega-6 and omega-3 fatty acids in plasma affect clotting activity; however, there was no relationship between the extent of liver haemorrhages and the composition of plasma fatty acids.

Concurrent use of veterinary drugs and herbal medicines in racing standardbreds.

Standardbred trainers from 1 racetrack and 7 off-track training facilities were surveyed to determine the most common drugs, and prevalence of concurrent herb administration. Furosemide (on-track) and anti-inflammatory drugs (off-track) were the most common drugs administered. Among horses on-track, 9.8% received herbs compared with 13.8% off-track horses; 67% and 58% of these horses, respectively, received concurrent drugs.

The effect of housing and methionine intake on hoof horn hemorrhages in primiparous lactating Holstein cows.

This study investigated the effects of housing and nutrition on the development of hoof horn disease (as identified by the appearance of hoof horn hemorrhages) in primiparous lactating Holstein cows. The first objective was to investigate whether replacing butyl rubber mats in cubicles (free stalls) with thicker mattresses filled with chopped rubber would significantly reduce hoof horn hemorrhages, and if this reduction would so affect the level of hoof horn hemorrhages as to make it similar to that observed in primiparous cows in straw yards. The second objective was to investigate the effect of methionine supplementation for the first 13 wk of lactation on the development of such hemorrhages. This study confirmed that both sole and white line hemorrhages increase during early lactation in housed cows, although the pattern of development of white line hemorrhages is not identical to that of sole hemorrhages. Housing primiparous cows in straw yards after calving significantly reduced the development of hoof horn hemorrhages, but replacing cubicle mats with thicker mattresses had no significant effect. Providing 115% of calculated methionine requirements had no significant impact on the development of hoof horn hemorrhages.

Long-term effects of feeding flaxseed on performance and egg fatty acid composition of brown and white hens.

Two hundred fifty-six 18-wk-old Shaver White and ISA-Brown pullets were fed commercial diets containing either 0 or 10% flaxseed in order to study the long-term effects of feeding flaxseed on hen performance and egg production parameters. Performance was monitored over 10 consecutive 28-d periods. Flaxseed was introduced gradually at 28 wk of age and was maintained until hens were 53 wk of age, when flaxseed was gradually eliminated from the diet. Feed intake was less (P < 0.05) for hens fed flaxseed compared to those consuming the control diet. Flax-fed hens were also lighter (P < 0.05) compared to the control birds. Egg production, egg weight, shell weight, albumen height, and shell thickness were not significantly (P > 0.05) different for hens consuming 0 and 10% flaxseed; however, yolk weight was reduced (P < 0.05) in hens fed flaxseed. Both strains of birds fed flaxseed deposited significantly more n-3 fatty acids into their eggs. Sampling of livers at the end of the trial showed that hens fed flaxseed had a higher (P < 0.05) incidence of liver hemorrhages.

Platelet function in dogs treated for lymphoma and hemangiosarcoma and supplemented with dietary n-3 fatty acids.

A prospective randomized, double-blind clinical trial was performed to test the hypothesis that dogs with malignancies that are supplemented with n-3 fatty acids do not have clinical or laboratory evidence of coagulation disorders or altered platelet function when compared with unsupplemented dogs with similar malignancies. Thirteen dogs with hemangiosarcoma and 66 dogs with lymphoma were evaluated. Coagulation status of the dogs with lymphoma and hemangiosarcoma was evaluated with prothrombin time, partial thromboplastin time, platelet count, and in vitro platelet aggregometry using the whole-blood method. These tests were performed at 5 time points: before beginning the diet (week 0), at weeks 3, 15, and 21, and at 1 year or when progressive disease was evident. Alterations in platelet function in dogs receiving a diet supplemented with dietary n-3 fatty acids were not identified when compared to dogs fed a control diet. Dietary n-3 fatty acid supplementation using this dosage and ratio in dogs with lymphoma or hemangiosarcoma did not induce clinically significant hemorrhage in these animals. Therefore, supplementation with n-3 fatty acids did not result in clinical or laboratory evidence relating to uncontrolled hemorrhage in these dogs.

Supplemental dietary biotin for prevention of lesions associated with aseptic subclinical laminitis (pododermatitis aseptica diffusa) in primiparous cows.

OBJECTIVE: To evaluate, in primiparous cows, the effect of dietary biotin supplementation on severity of lesions of aseptic subclinical laminitis. ANIMALS: 100 primiparous cows in a 800-cow commercial dairy herd. PROCEDURE: Cows were assigned after calving to a biotin-supplemented (20 mg/head/d) or control group on an alternating basis. Digits 3 and 4 of the left forelimb and right hind limb were examined at a mean of 25, 108, and 293 days after parturition. Toe length, hoof angle, and heel depth were measured, and hooves were examined for solear hemorrhage, yellow solear discoloration, separation of the white line, and heel erosion. Serum and milk biotin concentrations were also measured. RESULTS: Serum biotin concentrations were significantly higher for supplemented than for control cows. During the second examination, prevalence of separation of the white line of digits 3 and 4 of the hind limb and digit 4 of the forelimb was lower for supplemented than for control cows. Mean decrease in heel depth between the first and third examinations was approximately twice as great for digit 4 of the forelimb and 4 times as great for digit 3 of the forelimb in supplemented, versus control, cows. Other differences were not found. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that supplemental dietary biotin may have a beneficial effect on hoof health in intensively managed primiparous dairy cows.

Sweet clover poisoning in dairy cattle in California.

Eight of 600 Holstein heifers and cows died after ingestion of sweet clover silage (Melilotus sp) that contained excessive concentrations of dicumarol caused by mold infestation. The cattle developed subcutaneous hemorrhages and bled from the vagina, became weak, were unable to move, and died. To the best of our knowledge, this is the first report of sweet clover poisoning in cattle from California and is discussed in light of previous findings in the Midwest and Canada. Sweet clover poisoning is caused by dicumarol, a fungal metabolite produced from substrates in sweet clover, and is a common livestock problem in the Northern Plains and Canada. Sweet clover poisoning should be considered in livestock animals with clinical evidence of hemostatic dysfunction, prolonged coagulation times, subcutaneous hemorrhages, and hemorrhagic abortions. Definite diagnosis of moldy sweet clover poisoning can be accomplished by analysis of serum and feed samples for dicumarol concentrations.

Autotransfusion in the emergency patient.

As the need for blood products increases in the veterinary emergency setting, the technique of autologous transfusion is being re-explored. Analogous transfusions provide an immediate source of blood with known compatibility to the patient. With the advent of passive canister collection systems, the technique is becoming simplified and affordable for the veterinary community. Although complications such as coagulopathies and microemboli have been reported with the procedure, attention to patient selection, technique, and patient monitoring following transfusion greatly minimizes potential drawbacks.

Autologous blood instillation alters respiratory mechanics in horses.

To investigate physiological consequences of autologous blood instillation in the lungs of healthy horses, respiratory mechanics and bronchial response to histamine were studied in 8 Thoroughbreds before and after introducing autologous blood (n = 5) and sterile saline solution (n = 3) into their lungs. Blood instillation resulted in a decrease in dynamic compliance (Cdyn) and increased respiratory resistance (R). Bronchial sensitivity and reactivity were unchanged after blood introduction. There were no significant changes in pulmonary mechanics or bronchial response after saline instillation.