Effect of RNA interference with HIF-1α on the growth of pulmonary artery endothelial cells in broiler chickens.

Pulmonary artery remodeling is a characteristic feature of broiler ascites syndrome (BAS). Pulmonary artery endothelial cells (PAECs) regulated by HIF-1α play a critical role in pulmonary artery remodeling, but the underlying mechanisms of HIF-1α in BAS remain unclear. In this experiment, primary PAECs were cultured in vitro and were identified by coagulation factor VIII. After hypoxia and RNA interference, the mRNA and protein expression levels of HIF-1α and VEGF were determined by qPCR and Western blotting. The transcriptome profiles of PAECs were obtained by RNA sequencing. Our results showed that the positive rate of PAECs was more than 90%, hypoxia-induced promoted the proliferation and apoptosis of PAECs, and RNA interference significantly downregulated the expression of HIF-1α, inhibited the proliferation of PAECs, and promoted the apoptosis of PAECs. In addition, transcriptome sequencing analysis indicated that HIF-1α may regulate broiler ascites syndrome by mediating COL4A, vitronectin, vWF, ITGα8, and MKP-5 in the ECM, CAMs and MAPK pathways in PAECs. These studies lay the foundation for further exploration of the mechanisms of pulmonary artery remodeling, and HIF-1α may be a potentially effective gene for the prevention and treatment of BAS.

The effect of peroxisome proliferator-activated receptor gamma (PPARγ) as a mediator of dietary fatty acids and thiazolidinedione in pulmonary arterial hypertension induced by cold stress of broilers.

The objective of this experiment was to evaluate the effects of dietary fish oil and pioglitazone as peroxisome proliferators-activated receptor gamma (PPARγ) activating ligands on the reduction of cold-induced ascites in broiler chickens. A total of 480 one-day-old (Ross 308) male chicks were randomly allocated to four treatment groups with eight replicates of 15 birds each. The following treatments were used: 1) ambient temperature (negative control), with basal diet; 2) cold-induced ascites (positive control), with basal diet; 3) cold-induced ascites, with basal diet +10 mg/kg/day pioglitazone and 4) cold-induced ascites, with basal diet +1% of fish oil. When compared with the positive control, body weight gain was higher (P ≤ 0.05) for broilers fed diets containing fish oil and pioglitazone at 28, 42, and 0-42 d. Broilers under cold-induced ascites had the highest blood pressure at 21 and 42 d, while fish oil and pioglitazone treatment reduced the blood pressure (P ≤ 0.05). Red blood cells, white blood cells, hematocrit, erythrocyte osmotic fragility, bursa of Fabricius and spleen weights were improved (P ≤ 0.05) for chickens fed fish oil diets and pioglitazone compared to the cold-induced ascites (positive control). Exposure to cold temperature resulted in an increase in plasma T3 and T3/T4 ratio and decline in plasma T4 (P ≤ 0.05). In conclusion, PPARγ agonist pioglitazone and fish oil as source of omega-3 polyunsaturated fatty acid could be used as a strategy to reduce the negative effects of pulmonary arterial hypertension and ascites in broiler chickens.

Right congestive heart failure in domestic kitten (Felis domesticus) by dilation of the pulmonary veins and cardiomegali.

Background: Congestive heart failure (CHF) is a pathological state characterized by the incapability of the heart to properly perform its essential function of delivering blood to meet the metabolic demands of the body. Case Description: The present case report concerns a 3-month-old male domestic kitten, displaying symptoms including an enlarged abdomen, emaciation, dehydration, dyspnea, rhinorrhea, and infestation with scabies. This animal, weighing 0.7 kg displays a tabby bicolor pattern. The findings gleaned from the clinical evaluation revealed the presence of a murmur upon auscultation of the cardiac region. Upon conducting an ultrasound examination, it was determined that the abdominal cavity contained a fluid accumulation known as ascites. Conclusion: The findings from the radiographic evaluation indicate that feline Hiro exhibits ascites alongside cardiomegaly, in conjunction with discernible vascular modifications characterized by both enlargements of the pulmonary arteries and veins.

Budd-Chiari-like syndrome in a dog secondary to a gunshot wound treated with balloon angioplasty and endovascular stent placement.

A 6-year-old female spayed Chihuahua mix presented with chronic recurrent ascites. Computed tomographic angiography revealed an isolated stenosis of the caudal vena cava secondary to a metallic foreign body, resulting in Budd-Chiari-like syndrome. Balloon angioplasty and endovascular stent placement successfully resolved the obstruction with long-term resolution of ascites.

The effects of dietary inclusion of Camelina sativa on performance and ascites incidence in broilers subjected to cold exposure.

BACKGROUND: Camelina sativa oilseed has a high amount of polyunsaturated fatty acids (FAs) especially α-linolenic acid. n - 3 FA can improve the deformability of erythrocytes and mediate relaxations in coronary artery such as nitric oxide (NO) that is the principal vasodilator responsible for diminution the pulmonary arterial hypertension response. METHODS: To investigate the effects of camelina sources on ascites incidence in broilers kept on altitude, 672 male chicks were fed with 7 dietary treatments, including control, 2% or 4% camelina oil (CO), 5% or 10% camelina meal (CM) and 5% or 10% camelina seeds (CS) diets. RESULTS: Supplementation of 2% CO had no negative impact on performance, whereas feed intake and body weight gains decreased (p < 0.05) by adding 4% CO, CM and CS. Birds fed camelina diets had lower serum triglyceride at day 42, total cholesterol and LDL cholesterol at both 28 and 42 days. Plasma aspartate aminotransferase decreased (p < 0.001) among 5% and 10% CS groups on day 42. Serum and liver concentrations of malondialdehyde declined (p < 0.05) by camelina treatments, whereas serum NO and liver glutathione peroxidase activity significantly elevated. Red blood cell, heterophils and H:L ratio were lower but lymphocyte was higher in camelina groups. Inclusion of camelina reduced (p < 0.05) relative weights of heart and right ventricle, the weight of right ventricle to total ventricle ratio and ascites mortality. CONCLUSIONS: Use of 2% CO as a source of n - 3 FA can improve ascites condition and mortality in broilers reared at high altitude without the reduction of growth performance. However, feeding 4% CO or 5% and 10% CS or CM reduced broiler performance.

Peroxisome proliferator-activated receptor gamma (PPARγ) activation: a potential treatment for ascites syndrome in broiler chickens.

Ascites (serous fluid accumulation in the abdominal cavity) has been observed worldwide in fast growing broilers. Pulmonary vascular remodeling is an important pathological feature of broiler ascites syndrome. Peroxisome proliferators-activated receptor gamma (PPARγ) and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α) are expressed in pulmonary vascular endothelial cells and vascular smooth muscle cells (VSMC) where they participate in the regulation of normal pulmonary vascular function. The objective of the present study was to investigate the effects of omega-3 fatty acids (found in fish oil) and pioglitazone (PIO) as natural and synthetic PPARγ ligands supplementation on PPARγ and PGC-1α expression in the prevention of pulmonary arterial hypertension (PAH) syndrome in broiler chickens. The experiment was conducted with 4 treatment groups: 1) negative control, normal temperature conditions with basal diet; 2) positive control, low-temperature conditions with basal diet; 3) positive control + 10 mg PIO/kg of weight/d and 4) positive control + 1% FO. Each treatment had 5 replicates. Ascites heart index (RV/TV) was significantly (P < 0.05) reduced in chickens receiving FO (0.20) and PIO (0.21) compared to the positive control group (0.26). The addition of PIO in broilers under cold-induced ascites significantly increased the expression of PPARγ (9.44) and PGC-1α (5.81) genes in lung tissue compared to the negative control group (1.03, P < 0.05). Proliferative indexes of VSMC in pulmonary arteries such as PMT, PIT, and percentage wall thickness were significantly elevated in positive control group, indicating that pulmonary vascular remodeling occurred following VSMC proliferation in ascites. The vessel internal diameter was increased in FO and PIO groups. Based on these results, activation and expression of PPARγ and PGC-1α genes as a critical regulator of pulmonary artery smooth muscle cell using ligands, especially PIO, can be effective in reducing the incidence of PAH in broiler chickens.

N-terminal brain natriuretic peptide, cardiac troponin-I, and point-of-care ultrasound in dogs with cardiac and noncardiac causes of nonhemorrhagic ascites.

BACKGROUND: Nonhemorrhagic ascites (NHA) can be caused by cardiac diseases (cNHA) and noncardiac diseases (ncNHA). N-terminal brain natriuretic peptide (NT-proBNP), cardiac troponin-I (cTnI), and point-of-care ultrasound (POCUS) may differentiate between cNHA and ncNHA. HYPOTHESIS/OBJECTIVES: We compared NT-proBNP and cTnI concentrations as well as POCUS findings in dogs presented with cNHA and ncNHA. ANIMALS: Dogs (n = 60) were enrolled based on identification of NHA with an effusion packed cell volume < 10%. METHODS: Blood samples were collected and POCUS was performed on all dogs. Dogs were diagnosed with cNHA (n = 28) or ncNHA (n = 32) based on echocardiography. The cNHA group was subdivided into cardiac non-pericardial disease (n = 17) and pericardial disease (n = 11). RESULTS: The NT-proBNP concentration (median; range pmol/L) was significantly higher in the cNHA group (4510; 250-10 000) compared to the ncNHA group (739.5; 250-10 000; P = .01), with a sensitivity of 53.8% and specificity of 85.7% using a cut-off of 4092 pmol/L. The NT-proBNP concentrations were significantly higher in the cardiac non-pericardial disease group (8339; 282-10 000) compared with the pericardial disease group (692.5; 250-4928; P = .002). A significant difference in cTnI concentration (median; range ng/L) between the cNHA group (300; 23-112 612) and ncNHA group (181; 17-37 549) was not detected (P = .41). A significantly higher number of dogs had hepatic venous and caudal vena cava distension in the cNHA group compared to the ncNHA group, respectively (18/28 vs 3/29, P < .0001 and 13/27 vs 2/29, P < .001). Gall bladder wall edema was not significantly different between groups (4/28 vs 3/29, P = .74). CONCLUSIONS AND CLINICAL IMPORTANCE: NT-proBNP concentration and POCUS help distinguish between cNHA and ncNHA.

Preemptively planned en bloc resection of an extensive right adrenal pheochromocytoma involving the right hepatic division, caval thrombus and segmental caudal vena cava in a dog with Budd-Chiari-like syndrome.

BACKGROUND: Surgical resection is the treatment of choice for canine adrenal pheochromocytomas (PHEOs). Information on en bloc resection of adrenal PHEO with tumour thrombus, right hepatic division and segmental caudal vena cava (CVC) running through the adrenal tumour and right hepatic division is limited. OBJECTIVE: To describe the preemptively planned en bloc resection of an extensive right adrenal PHEO involving the right hepatic division, the caval thrombus and the segmental CVC in a dog with Budd-Chiari-like syndrome (BCLS). METHODS: A 13-year-old castrated male miniature dachshund was referred for surgical treatment due to anorexia, lethargy and severe abdominal distension caused by abundant ascites. Preoperative computed tomography (CT) revealed a large mass in the right adrenal gland with a large caval thrombus obstructing the CVC and hepatic veins, which caused BCLS. Additionally, collateral vessels were formed between the CVC and azygos veins. No findings suggested obvious metastases. Based on CT findings, an en bloc resection of the adrenal tumour with caval thrombus, right hepatic division and segmental CVC was planned. RESULTS: The preoperatively planned resection was feasible; the tumour was completely resected grossly. The operation time and total Pringle manoeuvre time were 162 min and 16 min 56 s, respectively. There was no postoperative hindlimb oedema, renal dysfunction, ascites or abdominal distention. The patient's clinical signs, including appetite, fully improved. Hospitalization lasted 16 days. However, the patient died on the 130th postoperative day due to suspected metastases and cachexia. CONCLUSIONS: Even in case of an extensive infiltration of adrenal PHEO causing BCLS, an en bloc resection might be successfully achieved based on the preoperative CT findings speculating the collateral vessels formed for caudal venous return.

Lymphomatosis as a Cause of Abdominal Pain and Distension in Two Adult Horses.

Two equine patients presented separately with severe abdominal distention, colic, lethargy, and decreased appetite. An ante-mortem diagnosis of lymphoma was reached in each case based on peritoneal fluid cytology. Due to a poor prognosis, the horses were humanely euthanized. Post-mortem examination with histology and immunohistochemistry confirmed both cases as lymphoma: alimentary B-cell lymphoma of the distal jejunum and cecum in one case, and T-cell lymphoma of the cecum in the second case. Both cases exhibited extensive metastasis with peritoneal and pleural serosae covered in small nodules and plaque like masses consistent with lymphomatosis. These cases document a unique presentation of lymphoma in equine patients presenting as peritoneal lymphomatosis with ascites.

Underlying mechanism of Qiling Jiaogulan Powder in the treatment of broiler ascites syndrome.

Broiler ascites syndrome (AS), is a nutritional and metabolic disease that occurs in fast-growing commercial broiler chickens. AS can cause poor growth and a significant increase in the rate of broiler deaths, which has resulted in serious economic losses to the poultry industry. The classic traditional Chinese medicine Qiling Jiaogulan Powder (QLJP) has been demonstrated to have a certain therapeutic effect on broiler AS. However, its pharmacological mechanism remains to be elucidated. This study was performed to investigate the multitarget action mechanism of QLJP in the treatment of broiler AS based on network pharmacology analysis using a broiler AS model. First, all chemical components and targets of QLJP were obtained from the Traditional Chinese Medicine System Pharmacology Analysis Platform (TCMSP). Targets related to broiler AS were further obtained through the GeneCards database and the NCBI Gene sub-database. A protein-protein interaction (PPI) network was constructed. Then, enrichment analyses were performed to predict the potential mechanisms of QLJP in the treatment of broiler AS. Finally, the treatment effect of QLJP on AS was verified in a broiler AS model. Network pharmacology analysis generated 49 active ingredients and 167 core targets of QLJP, and a QLJP-single drug-target-disease network was successfully constructed. Gene enrichment analysis indicated that the core targets have played major roles in the Cell cycle, FOXO signaling pathways, etc. We demonstrated that QLJP improved clinical and organ damage symptoms and significantly reduced the ascites heart index in broilers with AS induced by administration of high-energy, high-protein diets and high-sodium drinking water in a low-temperature environment. QLJP may regulate lung oxidative stress, the cell cycle and apoptosis by activating the FOXO3a signaling pathway to interfere with the occurrence and development of AS in broilers. QLJP administration may be a good clinical strategy for the prevention and treatment of broiler AS.

Managing broiler production challenges at high altitude.

This review covers the challenges of broiler chickens at high altitude, with the focus on growth performance and physiological response. The review also sheds light on nutritional and management interventions that help overcome the challenges raised at high altitude. Reduced concentration of atmospheric oxygen is by far the biggest challenge that remarkably affect growth performance and livability of broiler chickens reared in high altitude area. Broiler chickens have endured intensive genetic selection, which potentially predispose them to several metabolic disorders. Hypoxia is an overriding factor that may increase the incidence of metabolic disorders, mainly ascites syndrome at high altitude. Commercial broiler strains cannot fully achieve their genetic potential when raising at highland regions. Careful nutrition and management considerations are required to prevent metabolic disorders when raising broilers at high altitude. In ovo or in-feed nutraceuticals such as l-carnitine and guanidinoacetic acid as well as pharmaceuticals, texture of feed and the use of proper sources and levels of dietary energy and protein are important factors that need to be carefully considered for rearing broiler chickens at high altitude. Management strategies such as lighting programs have been shown to be effective to circumvent ascites prevalence. Special breeding programs may also be considered to develop strains with resistance to ascites.

Long-term palliation of right-sided congestive heart failure after stenting a recurrent cor triatriatum dexter in a 10½-year-old pug.

A 10½-year-old, male neutered, pug presented with increasing ascites over two months. Echocardiography revealed cor triatriatum dexter with no concurrent cardiovascular anomalies, subsequently confirmed by computed tomography angiography. Balloon dilation of the perforated intra-atrial membrane under fluoroscopic guidance resulted in the transient resolution of all clinical abnormalities, but six months later stenosis and ascites recurred. After repeated balloon dilation, a stent was placed across the membrane. The dog remains asymptomatic fourteen months after the second procedure. One noteworthy feature of this case is the onset of congestive heart failure due to a congenital defect only at more than 10 years of age.

Influence of dietary lycopene on growth performance, antioxidant status, blood parameters and mortality in broiler chicken with cold-induced ascites.

The aim of this study was to determine the effects of dietary lycopene supplementation on growth performance and antioxidant status of broiler chickens exposed to cold environment to induce ascites. Three hundred male chickens were exposed full-day to cold stress (CT, 10°C) starting from day 15 of age until the end of experiment at day 42, while a positive control group (NT, 100 birds) was kept under normal temperature (23-25°C). The CT groups (three treatments and five replicates of 20 birds) were as follows: negative control (basal diet, CT) and CT + 200 or 400 mg lycopene per kg diet from 15 to 42 d of age. Results showed that CT without lycopene supplementation caused a reduction of feed intake and weight gain and increased the feed conversion ratio. Supplementation of lycopene during CT restored the performance to levels of the positive control, lowered the index of right ventricles/total ventricles and ascites mortality. Birds reared under CT had lower serum activities of superoxide dismutase (SOD), glutathione peroxidase (GPx) and higher malondialdehyde (MDA) concentration than birds reared under the NT environment. With lycopene supplementation, serum MDA level significantly decreased and the activity of SOD and GPx increased. Blood concentration of haematocrit, haemoglobin and red blood cells were decreased by the highest lycopene supplementation to a level comparable to NT. Moreover, increasing dietary lycopene level suppressed serum concentrations of cholesterol and enhanced high-density lipoproteins levels in blood. In conclusion, lycopene supplementation alleviates adverse effects of cold stress on performance through modulating activity of antioxidant enzymes in broiler chickens.

Successful treatment of ascites accumulation and diarrhea associated with protein-losing enteropathy with oral equine placenta extract supplementation in a dog: A case report.

Background: Protein-losing enteropathy (PLE) is characterized by leakage of serum proteins into the intestinal lumen, indicating hypoproteinemia. Immunosuppressive agents are the mainstay of treatment, but in many cases, patients are forced to taper off early owing to the induction of liver damage. Case Description: An 8-year-old, non-spayed female Chihuahua presented with diarrhea and ascites effusion lasting 2 weeks. Based on the results of radiography and blood tests, a diagnosis of PLE was made. Prednisolone (3 mg/kg semel in die [SID]) and MitoMax (200 mg/day) were administered, but ascites accumulation and diarrhea did not improve. Thus, azathioprine (2 mg/kg/day) was added, but there was no improvement, and liver damage developed. The liver injury did not improve immediately, but diarrhea and ascites effusion improved after serum total protein and serum albumin levels increased after they had decreased. Subsequent tapering of prednisolone from 3 mg/kg SID to 1 mg/kg SID, combined with MitoMax (200 mg/day) and equine placenta extract (eqPE) (2 ml/day), resulted in no recurrence of ascites or diarrhea. Conclusion: In canine PLE with prolonged diarrhea and ascites effusion, supplementation with eqPE may be considered a reasonable additional therapeutic strategy.

Evaluation of the Effect of Peppermint Extract and Probiotics on Biochemical Factors in the Blood of Ascites-Induced Chickens.

Ascites syndrome occurs in growing broiler chickens in all parts of the world, which is one of the important causes of losses in many flocks, and its prevalence has been seen mostly in meat herds. The most important factor in the occurrence of ascites syndrome is the lack of oxygen in body tissues (hypoxia). Increasing the growth rate requires increasing the volume of blood flowing in the body to deliver nutrients to the organs and expel gases and metabolic products. Therefore, this experiment was conducted to compare the effect of peppermint extract and probiotics on the biochemical factors of the blood of chickens caused by ascites. The treatments were divided into 8 groups of 7 male chickens with 3 repetitions in each group at 21 days. The experimental treatments included control and treatments of peppermint, probiotic, peppermint and probiotic, induced ascites, induced ascites and peppermint, induced ascites and probiotics, induced ascites and peppermint and probiotics. At the end of 42 days, the blood factors of uric acid, triglyceride, glucose, cholesterol, ALT, ALP, and AST were measured by blood sampling. The experimental treatments significantly affected the investigated traits (P<0.05). Considering that in the treatment of induced ascites+ peppermint compared to the treatment of induced ascites, weight gain was significant, and in the blood factors of cholesterol, triglyceride, uric acid, glucose, and functional liver enzymes including ALT, AST, ALP recorded a significant decrease. Therefore, the effectiveness of peppermint extract in improving induced ascites in chickens was determined. Peppermint extract had a positive effect on induced ascites and improved the performance indicators of broiler chickens, and this extract can be used as a preventive of ascites.

Dysregulated expression of mRNA and SNP in pulmonary artery remodeling in ascites syndrome in broilers.

Broiler ascites syndrome (AS), also called pulmonary artery hypertension, is a metabolic disorder that has been observed worldwide in fast-growing broilers. Pulmonary arterial remodeling is a key step in the development of AS. The precise relationship between mRNA and SNP of the pulmonary artery in regulating AS progression remains unclear. In this study, we obtained pulmonary artery tissues from broilers with AS to perform pathologic section and pathologic anatomic observation. SNP, InDel, and mRNA data analysis were carried out using GATK and ANNOVAR software to study the SNP loci of 985 previously reported genes (437 upregulated and 458 downregulated). The pathology results showed that there was a lot of yellow fluid in the abdominal cavity and pericardium, that the ascites cardiac index and hematocrit changed significantly, and that the pulmonary artery had remodeled and become thicker in the disease group. Myocardial sections showed vacuolar degeneration of myocytes and rupture of muscle fibers. In addition, ALDH7A1, IRG1, GGT5, IGSF1, DHX58, USP36, TREML2, SPAG1, CD34, and PLEKHA7 were found to be closely associated with the pathogenesis of pulmonary artery remodeling in AS progression. Taken together, our present study further illuminates the molecular mechanism of pulmonary artery remodeling underlying AS progression.

Use of immune repertoire sequencing to resolve discordant microscopic and immunochemical findings in a case of T cell-rich large B cell lymphoma in a young dog.

BACKGROUND: Lymphocytic neoplasms with frequent reactive lymphocytes are uncommonly reported in dogs, and can pose a diagnostic challenge. Different diagnostic modalities such as cytology, flow cytometry, histopathology, immunohistochemistry, and clonality testing, are sometimes required for a diagnosis. This report illustrates the value of using a multi-modal diagnostic approach to decipher a complex lymphocytic tumor, and introduces immune repertoire sequencing as a diagnostic adjunct. CASE PRESENTATION: A 10-month-old Great Dane was referred for marked ascites. Cytologic analysis of abdominal fluid and hepatic aspirates revealed a mixed lymphocyte population including numerous large lymphocytes, yielding a diagnosis of lymphoma. Flow cytometrically, abdominal fluid lymphocytes were highly positive for CD4, CD5, CD18, CD45, and MHC II, consistent with T cell lymphoma. Due to a rapidly deteriorating clinical condition, the dog was euthanized. Post mortem histologic evaluation showed effacement of the liver by aggregates of B cells surrounded by T cells, suggestive of hepatic T cell-rich large B cell lymphoma. Immune repertoire sequencing confirmed the presence of clonal B cells in the liver but not the abdominal fluid, whereas reactive T cells with shared, polyclonal immune repertoires were found in both locations. CONCLUSIONS: T cell-rich large B cell lymphoma is a rare neoplasm in dogs that may be challenging to diagnose and classify due to mixed lymphocyte populations. In this case, the results of histopathology, immunohistochemistry and immune repertoire sequencing were most consistent with a hepatic B cell neoplasm and reactive T cells exfoliating into the abdominal fluid. Immune repertoire sequencing was helpful in delineating neoplastic from reactive lymphocytes and characterizing repertoire overlap in both compartments. The potential pitfalls of equating atypical cytomorphology and monotypic marker expression in neoplasia are highlighted.

Disseminated Mycobacterium avium subsp. hominissuis infection and ascites in an FIV-positive cat.

A domestic shorthair cat was presented to the Veterinary Teaching Hospital at The University of the West Indies with a history of anorexia, ataxia, and lethargy. On physical examination, moderate abdominal distension and a palpable abdominal fluid wave were noted. Dark yellow, cloudy fluid was collected via abdominocentesis. Fluid analysis indicated that the effusion was a transudate containing low numbers of macrophages and occasional neutrophils. Some of the macrophages contained rod-shaped nonstaining structures of variable length (2-4 um). These structures were also seen extracellularly in low numbers. The morphology of the structures was suggestive of Mycobacterium. The cat's condition continued to deteriorate, and it died within a few hours of being admitted. Further diagnostic tests revealed feline immunodeficiency virus (FIV) infection with concurrent Mycobacterium avium subsp hominissuis infection. To the authors' knowledge, this is the first reported case of nontubercular mycobacterial-related ascites in a cat.

Identification and validation of quantitative trait loci for ascites syndrome in broiler chickens using whole genome resequencing.

BACKGROUND: Ascites syndrome is a hypertensive, multifactorial, multigene trait affecting meat-type chickens imposing significant economic losses on the broiler industry. A region containing the CPQ gene has been previously identified as significantly affecting ascites phenotype. The region was discovered through whole genome resequencing focused on chicken chromosome 2. The association was confirmed through further genotyping in multiple broiler populations. RESULTS: The whole genome resequencing analyses have now been extended to the current chicken genome assembly. DNA samples were pooled according to gender and phenotype and the pools subjected to next generation sequencing. Loci were identified as clusters of single nucleotide polymorphisms where frequencies of the polymorphisms differed between resistant and susceptible chickens. The chickens are an unselected line descended from a commercial elite broiler line. Regions identified were specific to one or both genders. The data identify a total of 28 regions as potential quantitative trait loci for ascites. The genes from these regions have been associated with hypertensive-related traits in human association studies. One region on chicken chromosome 28 contains the LRRTM4 gene. Additional genotyping for the LRRTM4 region demonstrates an epistatic interaction with the CPQ region for ascites phenotype. CONCLUSIONS: The 28 regions identified were not previously identified in a multi-generational genome wide association study using 60k Single Nucleotide Polymorphism panels. This work demonstrates the utility of whole genome resequencing as a cost effective, direct, and efficient method for identifying specific gene regions affecting complex traits. The approach is applicable to any organism with a genome assembly and requires no a priori assumptions.

Characterization of ascites-derived aldehyde dehydrogenase-positive ovarian cancer stem cells isolated from Leghorn chickens.

Leghorn chickens are used as a preclinical model of ovarian cancer as they develop epithelial ovarian adenocarcinoma spontaneously at a very high frequency. Ovarian cancer is the most lethal disease among all gynecological malignancies in women. A small proportion of ovarian cancer stem cells are responsible for drug resistance and relapse of ovarian cancer. The objectives of this study are to isolate ovarian cancer stem cells from ascites of Leghorn chickens that spontaneously developed ovarian cancer and to determine their invasiveness, spheroid formation in three-dimensional culture devoid of extracellular matrix over several months. Ovarian cancer cells obtained from ascites were subjected to ALDEFLOUR assay that measures aldehyde dehydrogenase (ALDH) activity to separate ALDH1+ and ALDH1- cells by fluorescence-activated cell sorting. The cells were cultured using serum-free media for up to 6 mo in ultra-low attachment plates. Invasiveness of ALDH1+ and ALDH1- cells was determined by Matrigel invasion assay. Cellular uptake of acetylated low-density lipoprotein was evaluated. A small proportion (<4.75%) of ovarian cancer cells isolated from ascites were found to be ALDH1+ cells. ALDH1+ cells formed a greater number of spheroids and were also highly invasive in extracellular matrix compared to ALDH1- cells. Several spheroids developed 0.1- to 1-mm-long capillary-like tubules connecting other spheroids, thus forming a complex network that underwent remodeling over several months. Cells in the spheroids incorporated acetylated low-density lipoprotein suggestive of scavenger receptor activity. In summary, ALDH1+ ovarian cancer stem cells isolated from ascites of chickens appear to be invasive and form spheroids with complex networks of tubules reminiscent of vascular mimicry. Understanding the structure and function of spheroids and tubular network would provide valuable insight into the biology of ovarian cancer and improve poultry health.