Differential expression of fatty acid transporters and fatty acid synthesis-related genes in crop tissues of male and female pigeons (Columba livia domestica) during incubation and chick rearing

1. The growth performance of squabs reared solely by male or female parent pigeons was measured, and the changes of lipid content of crop milk and the expression profiles of genes potentially involved in lipid accumulation by crop tissues of parent pigeons were evaluated during incubation and chick rearing.

2. Squabs increased in body weight during 25 d of rearing, whereas both male and female pigeons lost weight after finishing rearing chicks, and the weight loss of male pigeons was significantly greater than that of female parent pigeons. Lipid content of crop milk from both parent pigeons gradually decreased to the crude fat level in the formulated diet after 10 d (R10) of chick rearing.

3. The gene expression of fatty acid translocase (FAT/CD36), fatty acid-binding protein 5 (EFABP) and acyl-CoA-binding protein (ACBP) in male pigeon crop tissue were the greatest at 17 d (I17) of incubation. In female pigeons, FAT/CD36 expression was the highest at I14, and both EFABP and ACBP expression peaked at I14 and R7. The expression of acetyl-CoA carboxylase and fatty acid synthase in male pigeons reached the maximum level at R1, while they peaked at I14 and I17, respectively in female pigeons. The gene expression of peroxisome proliferators-activated receptor-gamma (PPARγ) was the greatest at I17 in the male, while it was at I14 in the female. However, no regular changing pattern was found in PPARα gene expression in male pigeons.

4. These results indicated that male and female pigeons may make different contributions in rearing squabs. The gene expression study suggested that fatty acids used in lipid biosynthesis of crop milk probably originated from both exogenous supply and de novo synthesis. The sex of the parent pigeon affected the lipid content of crop milk and the expression profiles of genes involved in fatty acid transportation and lipogenesis.     

Antimicrobial Resistance Genes in Pigeons from Public Parks in Costa Rica

Antimicrobial resistance is known to be an emerging problem, but the extent of the issue remains incomplete. The aim of this study was to determine the presence or absence of nine resistance genes (bla_TEM, catI, mecA, qnrS, sulI, sulII, tet(A), tet(Q), vanA) in the faeces of 141 pigeons from four urban parks in Alajuela, Guadalupe, Tres Ríos and San José in Costa Rica. The genes were identified by real‐time PCR directly from enema samples. About 30% of the samples were positive for genes catI and sulI; between 13% and 17% were positive for qnrS, sulII, tet(A) and tet(Q); and 4% were positive for bla_TEM. The mecA and vanA genes were not detected. The average of antimicrobial resistance genes detected per pigeon was 2. Eight different patterns of resistance were identified, without differences in the sampling areas, being the most common pattern 2 (sulII positive samples). During rainy season, the genes more frequently found were sulI and tet(A). In conclusion, the urban inhabiting pigeons tested are currently carrying antimicrobial resistance genes, potentially acting as reservoirs of resistant bacteria and vectors to humans. To the authors’ knowledge, this is the first study carried out on direct detection of resistance genes in the digestive metagenomes of pigeons.     

Dietary apple polyphenols supplementation enhances antioxidant capacity and improves lipid metabolism in weaned piglets

Apple polyphenols (APPs) are biologically active flavonoids that have antioxidant, anti‐inflammatory, improving insulin sensitivity, hypocholesterolaemic effect and antiviral properties. This study was conducted to explore effects of dietary APPs supplementation on antioxidant activities and lipid metabolism in weaned piglets. Fifty‐four weaned piglets (half male and female) were randomly divided into three groups with six replicates in each group and three piglets in each repetition. Piglets were fed control diet (basal diet) or a control diet supplemented with 400 mg/kg or 800 mg/kg APPs for 6 weeks. Blood and liver samples were collected to determine biochemical, antioxidant and lipid metabolism parameters. Here we showed that dietary APPs supplementation increased HDL‐C and decreased T‐CHO, TG and LDL‐C concentrations. Dietary APPs supplementation increased antioxidative capacity in serum and CAT activity in liver, and significantly increased the mRNA expressions of CAT, GST and SOD1 in liver. ACC mRNA level and LPL activity were tended to decrease by APPs. HMG‐CoAR, CTP7A1, CD36 and FATP1 mRNA levels were decreased by APPs, while LDL‐R, PGC‐1α, Sirt1 and CPT1b mRNA levels were increased by 400 mg/kg APPs. No alterations in growth performance were found in all treatments. This study firstly provided the evidence that dietary APPs supplementation could enhance systemic antioxidant capacity and improve lipid metabolism in weaned piglets. The mechanism by which APPs improve lipid metabolism might be through regulating hepatic cholesterol metabolism and increasing fatty acid oxidation, and decreasing fatty acid uptake and de novo synthesis.     

High‐level exogenous trans10, cis12 conjugated linoleic acid plays an anti‐lipogenesis role in bovine mammary epithelial cells

Primary bovine mammary epithelial cells (BMECs) were treated by 0, 37.5, 75, 112.5, 150 μmol/L trans10, cis12 conjugated linoleic acid (CLA) to evaluate the effects of different level trans10, cis12 CLA on lipogenesis in BMEC. Addition of 75–150 μmol/L trans10, cis12 CLA reduced significantly the triacylglycerol (TAG) content (P < 0.05), but did not have inhibiting action on cell proliferation (P > 0.05). Treatment with 150 μmol/L trans10, cis12 CLA for 48 h resulted in a 17.1% reduction (P < 0.0001) of medium chain fatty acids (MCFA, C14 < C < C16), a 26.5% reduction (P < 0.0001) of unsaturated fatty acids (UFA) and a corresponding reduction of the mRNA abundance of acetyl coenzyme A (acetylCoA) carboxylase (ACC) (P = 0.046), fatty acid synthase (FAS) (P = 0.017) and stearoylCoA desaturase1 (SCD1) (P = 0.002). Another finding was that trans10, cis12 CLA elevated expression of diacylglycerol acyltransferase2 (DGAT2) (P = 0.020) and long chain acylCoA synthetases (ACSL) (P = 0.032). In conclusion, higher trans10, cis12 CLA, not low trans10, cis12 CLA, inhibited milk fat synthesis and changed fatty acid composition by regulating the expression of FAS, ACC, SCD1, DGAT2 and ACSL.     

Effect of sorghum grain supplementation on glucose metabolism in cattle and sheep fed temperate pasture

The aim of this work was to evaluate the effect of sorghum grain supplementation on plasma glucose, insulin and glucagon concentrations, and hepatic mRNA concentrations of insulin receptor (INSR), pyruvate carboxylase (PC), and phosphoenolpyruvate carboxykinase (PCK1) mRNA and their association with nutrient intake, digestion and rumen volatile fatty acids (VFA) in cattle and sheep fed a fresh temperate pasture. Twelve Hereford × Aberdeen Angus heifers and 12 Corriedale × Milchschaf wethers in positive energy balance were assigned within each species to one of two treatments (n = 6 per treatment within specie): non‐supplemented or supplemented with sorghum grain at 15 g/kg of their body weight (BW). Supplemented cattle had greater plasma glucose concentrations, decreased plasma glucagon concentrations and tended to have greater plasma insulin and insulin‐to‐glucagon ratio than non‐supplemented ones. Hepatic expression of INSR and PC mRNA did not differ between treatments but PCK1 mRNA was less in supplemented than non‐supplemented cattle. Supplemented sheep tended to have greater plasma glucagon concentrations than non‐supplemented ones. Plasma glucose, insulin, insulin‐to‐glucagon ratio, and hepatic expression of INSR and PC mRNA did not differ between treatments, but PCK1 mRNA was less in supplemented than non‐supplemented sheep. The inclusion of sorghum grain in the diet decreased PCK1 mRNA but did not affect PC mRNA in both species; these effects were associated with changes in glucose and endocrine profiles in cattle but not in sheep. Results would suggest that sorghum grain supplementation of animals in positive energy balance (cattle and sheep) fed a fresh temperate pasture would modify hepatic metabolism to prioritize the use of propionate as a gluconeogenic precursor.     

Effect of chicken leptin recptor short hairpin RNA on expression of JAK2, STAT3, SOCS3 and CPT1 genes in chicken preadipocytes

In this study, we detect depressive effect on leptin receptor (LEPR) by LEPR‐specific short hairpin RNA (shRNA) expression plasmids in chicken preadipocytes, and effect on messenger RNA (mRNA) expression levels of genes related to signal transduction, including JAK2, STAT3, SOCS3 as well as CPT1, which is associated with fatty acid metabolism. shRNA expression vectors targeting LEPR were constructed and transfected into chicken preadipocytes. The transfection efficiency was evaluated by fluorescence microscopy. Real‐time PCR was used to detect its effect on mRNA expression levels of JAK2, STAT3, SOCS3 and CPT1. Results showed that LEPR mRNA was knocked down by 99% (P < 0.01) after transfection for 72 h. In the knockdown preadipocytes, the mRNA levels of JAK2 and CPT1 were down‐regulated by 47.56% (P < 0.01) and 42.26% (P < 0.05), respectively; while expression of STAT3 and SOCS3 increased 7.72‐fold (P < 0.01), 1.71‐fold (P < 0.01), respectively. It is concluded that knockdown of LEPR influences mRNA expression of its down‐stream genes, suggesting that chicken LEPR play a certain role in regulating genes in the complicated gene network of preadipocytes.     

A single‐nucleotide polymorphism in the canine cytochrome b5 reductase (CYB5R3) gene is associated with sulfonamide hypersensitivity and is overrepresented in Doberman Pinschers

Canine sulfonamide hypersensitivity (HS) has been associated with a variant in the cytochrome b₅ reductase gene (CYB5R3 729A>G), which encodes a drug‐detoxifying enzyme. Study objectives were to determine variant allele frequency in Doberman Pinschers (DOBE), a breed which may be predisposed to sulfonamide HS, and to characterize the effects of CYB5R3 729G on gene expression and function. CYB5R3 729A>G allele frequencies were compared between DOBE (n = 24) vs. non‐Doberman (non‐DOBE; n = 60) dogs. CYB5R3mRNA expression, protein expression, and reduction of sulfamethoxazole hydroxylamine were compared between banked canine liver samples of 729AA vs. GG genotype. The 729G allele was overrepresented in DOBE (1.00) vs. non‐DOBE dogs (0.567, p < .0001). mRNA and protein expressions as well as cyt b₅ reductase activity were similar between livers of AA and GG genotype. All Doberman Pinschers in this study were homozygous for CYB5R3 729G, which could contribute to this breed's apparent predisposition to sulfonamide HS. However, CYB5R3 729G does not alter sulfamethoxazole detoxification capacity, so a direct role could not be demonstrated. It is possible that this marker is linked to another contributing variant.     

Differential expression of genes implicated in liver lipid metabolism in broiler chickens differing in weight

ABSTRACT

1. Lipid parameters and expression of ACACA, APOA1, CPT1A, FASN, FOXO1, LIPG, PPARα and SIRT1 genes involved in lipid metabolism were investigated in two groups of high (HW) and low (LW) weight broilers from the same strain.

2. Blood cholesterol and liver triglyceride levels were significantly increased in HW chickens compared to LW broilers, while other parameters, i.e. blood triglyceride, blood HDL/LDL, liver cholesterol and total liver fat showed no significant changes in either group.

3. The relative expression of ACACA, APOA1 and CPT1A genes was significantly lower in the liver tissues of HW broilers than in the LW group. The mRNA levels of these three genes showed a significant negative correlation with abdominal fat deposition and live weight of broilers. However, relative expression of FASN, FOXO1, LIPG, PPARα and SIRT1 hepatic genes did not differ among broilers.

4. It was concluded that, of eight hepatic genes implicated in lipid metabolism, only the expression of three (ACACA, APOA1 and CPT1A) were significant for fat and leanness within the same strain of chicken. Since reducing body fat is a major goal in the broiler industry, these data can provide fresh insight into the molecular processes underlying the regulation of fat deposition in broilers.     

Expression of genes related to liver fatty acid metabolism in fat‐tailed and thin‐tailed lambs during negative and positive energy balances

Fat‐tailed sheep breeds can tolerate periods of negative energy balance without suffering from elevated concentration of plasma non‐esterified fatty acid (NEFA). This ability was attributed to unique metabolism of fat‐tailed adipose depot, whereas role of liver as an influential organ in fatty acid metabolism was not evaluated yet. Hence, current study was conducted to evaluate the effects of negative and positive energy balances on liver expression of genes related to fatty acid metabolism in fat‐tailed and thin‐tailed lambs. Lambs experienced negative (21 days) and positive (21 days) energy balances and were slaughtered at the beginning and end of negative energy balance and at the end of positive energy balance. Real‐time quantitative polymerase chain reaction (RT‐Q‐PCR) was conducted to evaluate changes in gene expression. Expression of diglyceride acyltransferase 1 (DGAT1), 3‐hydroxy‐3‐methylglutaryl‐CoA synthase 2 (HMGCS2) and apolipoprotein B (APOB) was not affected by genotype, energy balance and their interaction. Expression of carnitine palmitoyltransferase 1 (CPT1) was significantly higher in liver of fat‐tailed comparing to thin‐tailed lambs regardless of energy balance (p < 0.02). Catalase mRNA abundance was increased in response to negative energy balance (p < 0.02), and severity of this enhancement was higher in fat‐tailed lambs (p < 0.06). Expression of CPT1 was positively correlated with expression of HMGCS2 in both fat‐tailed (p < 0.05) and thin‐tailed lambs (p < 0.002); however, the correlation was weaker in fat‐tailed lambs (0.72 vs. 0.57, respectively, for thin‐tailed and fat‐tailed lambs). There was a positive correlation between DGAT1 and APOB genes expression in fat‐tailed lambs (0.94; p < 0.001), whereas this correlation was not observed in thin‐tailed lambs. Results demonstrate that liver of fat‐tailed lambs has higher capacity for metabolism of mobilized NEFA exposed to liver during negative energy balance.     

Diversity of Plasmids and Antimicrobial Resistance Genes in Multidrug‐Resistant Escherichia coli Isolated from Healthy Companion Animals

The presence and transfer of antimicrobial resistance genes from commensal bacteria in companion animals to more pathogenic bacteria may contribute to dissemination of antimicrobial resistance. The purpose of this study was to determine antimicrobial resistance gene content and the presence of genetic elements in antimicrobial resistant Escherichia coli from healthy companion animals. In our previous study, from May to August, 2007, healthy companion animals (155 dogs and 121 cats) from three veterinary clinics in the Athens, GA, USA area were sampled and multidrug‐resistant E. coli (n = 36; MDR, resistance to ≥2 antimicrobial classes) were obtained. Of the 25 different plasmid replicon types tested by PCR, at least one plasmid replicon type was detected in 94% (34/36) of the MDR E. coli; four isolates contained as many as five different plasmid replicons. Nine replicon types (FIA, FIB, FII, I2, A/C, U, P, I1 and HI2) were identified with FIB, FII, I2 as the most common pattern. The presence of class I integrons (intI) was detected in 61% (22/36) of the isolates with eight isolates containing aminoglycoside‐ and/or trimethoprim‐resistance genes in the variable cassette region of intI. Microarray analysis of a subset of the MDR E. coli (n = 9) identified the presence of genes conferring resistance to aminoglycosides (aac, aad, aph and strA/B), β‐lactams (ampC, cmy, tem and vim), chloramphenicol (cat), sulfonamides (sulI and sulII), tetracycline [tet(A), tet(B), tet(C), tet(D) and regulator, tetR] and trimethoprim (dfrA). Antimicrobial resistance to eight antimicrobials (ampicillin, cefoxitin, ceftiofur, amoxicillin/clavulanic acid, streptomycin, gentamicin, sulfisoxazole and trimethoprim‐sulfamethoxazole) and five plasmid replicons (FIA, FIB, FII, I1 and I2) were transferred via conjugation. The presence of antimicrobial resistance genes, intI and transferable plasmid replicons indicate that E. coli from companion animals may play an important role in the dissemination of antimicrobial resistance, particularly to human hosts during contact.     

Epidemiology of Chlamydia psittaci Infection in Racing Pigeons and Pigeon Fanciers in Beijing,China

Over 3 million racing pigeons (Columba livia) are registered in Beijing City Center for gambling purposes. During 2008–2010, we evaluated the occurrence and prevalence of Chlamydia psittaci in racing pigeons as well as the possible zoonotic transmission to pigeon fanciers in six districts of Beijing where pigeon races are particularly popular. C. psittaci‐specific serum antibody titres were obtained from 370 pigeons and 79 fanciers using enzyme‐linked immunosorbent assay. In addition, 206 and 67 throat swabs were, respectively, collected from pigeons and fanciers and tested for the presence of chlamydial antigen using immunofluorescence. C. psittaci‐specific serum antibody was detected in 37 of 370 pigeons and 19 of 79 fanciers. Of 206 pigeon clinical specimens, 55 were positive for C. psittaci antigen, while 16 of 67 swabs from the pigeon fanciers were positive. Based on ompA sequence analysis, the genotype of several avian and human isolates was genotype B. Thus, both high‐titre C. psittaci‐specific antibody and C. psittaci‐specific antigen were found with relatively high frequency in the pigeon flocks as well as in the pigeon fanciers. Our study suggests that C. psittaci infection is prevalent among the racing pigeon population in Beijing. Moreover, detection of serum antibodies and antigen in pigeon fanciers suggests that exposure and possible zoonotic transmission of C. psittaci from racing pigeons to humans does occur. In view of the life‐threatening respiratory illness C. psittaci may cause in humans, regulatory public health measures, to prevent further spread of the pathogen in avian populations and possible transmission to exposed humans, are urgently needed.     

Growth of rumen papillae in weaned calves is associated with lower expression of insulin‐like growth factor‐binding proteins 2, 3, and 6

This study aimed to characterize the relationship between the growth of rumen papillae in calves and the mRNA expression of insulin‐like growth factor‐binding proteins (IGFBPs) in the rumen papillae. The length of rumen papillae, the mRNA expression of IGFBPs in rumen papillae by quantitative real‐time PCR, and the presence of insulin‐like growth factors I and II (IGF‐I and II) by immunohistochemistry (IHC) were analyzed in nine Holstein calves divided into three groups: suckling (2 weeks, n = 3), milk‐continued (8 weeks, n = 3), and weaned (8 weeks, n = 3). The length of rumen papillae was greater (p < 0.01) in weaned calves than in suckling and milk‐continued calves, whereas the expressions of IGFBP2, IGFBP3, and IGFBP6 genes were lower (p < 0.05) in the rumen papillae of weaned calves than in milk‐continued calves. Thus, rumen papillae length and IGFBP2, 3, and 6 expressions were negatively correlated. The IHC analysis showed that IGF‐I and IGF‐II were present in the rumen epithelium of calves. These results suggested that the growth of rumen papillae after weaning is associated with the induction of IGFs by the low levels of IGFBP2, IGFBP3, and IGFBP6.     

In ovo carbohydrate supplementation modulates growth and immunity‐related genes in broiler chickens

A study was undertaken to investigate the role of in ovo administrated carbohydrates on the expression pattern of growth and immune‐related genes. In ovo injections (n = 400) were carried out on the 14th day of incubation into the yolk sac/amnion of the broiler chicken embryos. Expression of growth‐related genes: chicken growth hormone (cGH), insulin‐like growth factor‐I & II (IGF‐I & II) and mucin were studied in hepatic and jejunum tissues of late‐term embryo and early post‐hatch chicks. Expression of candidate immune genes: Interleukin‐2, 6, 10 and 12 (IL‐2, IL‐6, IL‐10 and IL‐12), Tumour necrosis factor‐alpha (TNF‐α) and Interferon gamma (IFN‐γ) were studied in peripheral blood monocyte cells of in ovo‐injected and control birds following antigenic stimulation with sheep RBC (SRBC) or mitogen concanavalin A (Con‐A). Glucose injection significantly increased the expression of IGF‐II gene during embryonic period and both cGH and IGF‐II in early post‐hatch period, while ribose‐injected chicks had higher expression of IGF‐II gene during embryonic stage. Enhanced mucin gene expression was also observed in fructose‐injected chicks during embryonic age. Glucose‐injected chicks had higher expression of IL‐6 or IL‐10, while those injected with fructose or ribose had higher expression of IL‐2, IL‐12 and IFN gamma. It is concluded that in ovo supplementation of carbohydrates might help in improving the growth of late‐term embryos and chicks. In ovo glucose could modulate humoral‐related immunity, while fructose or ribose might help in improving the cellular immunity in broiler chickens.     

Gossypol inhibits LH‐induced steroidogenesis in bovine theca cells

Gossypol, a polyphenolic aldehyde found in cottonseed, has been shown to perturb steroidogenesis in granulosa and luteal cells of rats, pigs and cattle. However, little is known about the direct effect of gossypol on theca cell functions in any species. The present study was conducted to investigate the effect of gossypol on the steroidogenesis and the expression of genes involved in it in cultured bovine theca cells. Theca cells were isolated from healthy preovulatory follicles and were cultured in the presence of luteinizing hormone (LH) for up to 7 days. During the culture period, main steroid products of the theca cells shifted from androstenedione (A4) at day 1 to progesterone (P4) from day 2 onward. At days 1 and 7, theca cells were treated with gossypol (0‐25 μg/mL) for 24 h. Gossypol inhibited LH‐stimulated theca cell A4 and P4 production in a dose‐dependent manner at both occasions. The viability of theca cells was not affected by gossypol at any doses used. Gossypol down‐regulated expressions of steroidogenic enzymes CYP11A1, HSD3B1 and CYP17A1, but not that of LHR. These results indicate that gossypol inhibits thecal steroidogenesis through down‐regulating gene expressions of steroidogenic enzymes but without affecting cell viability in cattle.     

Polymorphism of GDF9 and BMPR1B genes and their association with litter size in Markhoz goats

The main objective of this study was to investigate the polymorphism of GDF9 and BMPR1B genes and their relationship with litter size in Markhoz goats. The polymorphism of GDF9 and BMPR1B genes as well‐documented genes regarding fecundity in sheep and goat was investigated using RFLP‐PCR and a tetra‐primer amplification refractory mutation system‐PCR (T‐ARMS‐PCR) in Markhoz goats. The 164 blood samples were collected from the raised goats in Sanandaj Markhoz goat Performance Testing Station. The DNA extraction was carried out by salting‐out procedure, and then, PCR was performed using four and two pairs of primers to detect polymorphism in GDF9 and BMPR1B genes, respectively. To disclose GDF9 loci polymorphism, PCR products were digested with SspI (G3288A), PvuII (G423A), MvaI (A959C) and MspI (G1189A) restriction enzymes. The results showed that these mutations are available in tested animals. Parity had no significant effect on litter size. Also, the effects of different genotypes of GDF9 and BMPR1B had no significant effect on litter size. Further studies with a high number of animals with minimum relatedness for testing the association of these SNPs and others in the fecundity genes with reproductive traits may be worthwhile.     

Multidrug-resistant Proteus mirabilis isolates carrying blaOXA-1 and blaNDM-1 from wildlife in China: increasing public health risk

The emergence of multidrug resistance (MDR) in Proteus mirabilis clinical isolates is a growing public health concern and has serious implications for wildlife. What is the role of wildlife has been become one of the hot issues in disseminating antimicrobial resistance. Here, 54 P. mirabilis isolates from 12 different species were identified. Among them, 25 isolates were determined to be MDR by profile of antimicrobial susceptibility; 10 MDR P. mirabilis isolates were subjected to comparative genomic analysis by whole genome sequencing. Comprehensive analysis showed that chromosome of P. mirabilis isolates mainly carries multidrug-resistance complex elements harboring resistance to carbapenem genes bla_OXA-1, bla_NDM-1, and bla_TEM-1. Class I integron is the insertion hotspot of IS26; it can be inserted into type I integron at different sites, thus forming a variety of multiple drug resistance decision sites. At the same time, Tn21, Tn7, and SXT/R391 mobile elements cause widespread spread of these drug resistance genes. In conclusion, P. mirabilis isolates from wildlife showed higher resistance to commonly used clinic drugs comparing to those from human. Therefore, wild animals carrying MDR clinical isolates should be paid attention to by the public health.     

Epistatic effects between pairs of the growth hormone secretagogue receptor 1a,growth hormone,growth hormone receptor,non‐SMC condensin I complex,subunit G and stearoyl‐CoA desaturase genes on carcass,price‐related and fatty acid composition traits in Japanese Black cattle

Growth hormone secretagogue receptor 1a (GHSR1a), growth hormone (GH), growth hormone receptor (GHR), non‐SMC condensin I complex, subunit G (NCAPG) and stearoyl‐CoA desaturase (SCD), are known to play important roles in growth and lipid metabolisms. Single and epistatic effects of the five genes on carcass, price‐related and fatty acid (FA) composition traits were analyzed in a commercial Japanese Black cattle population of Ibaraki Prefecture. A total of 650 steers and 116 heifers for carcass and price‐related traits, and 158 steers for FA composition traits were used in this study. Epistatic effects between pairs of the five genes were found in several traits. Alleles showing strain‐specific differences in the five genes had significant single and epistatic effects in some traits. The data suggest that a TG‐repeat polymorphism of the GHSR1a.5′UTR‐(TG)_n locus plays a central role in gene–gene epistatic interaction of FA composition traits in the adipose tissue of Japanese Black cattle.     

Co-culture model reveals the characteristics of theca cells and the effect of granulosa cells on theca cells at different stages of follicular development

Theca cells (TCs) play an important role in follicular development, which cannot be separated from granulosa cells (GCs). However, compared with mammals, the TCs and the effects of GCs on TCs at different follicular development stages (FDSs) have specific characteristics in avian species, but none of them have been clearly defined. In this study, we established an in vitro co-culture (with GC at the corresponding stage) model of goose TCs at different FDSs (pre-hierarchical, hierarchical and F1) by using a transwell system. The properties of TCs in co-culture at the three FDSs, including cell morphology, activity and intracellular lipid content, as well as the expression of key genes involved in de novo lipogenesis, steroidogenesis, proliferation and apoptosis, were examined and defined. We further compared the mono-culture and co-culture groups. After co-culture, the activity of TCs showed significant (p < .01) increases in all stages; moreover, in pre-hierarchical TCs, the expression levels of FAS, SREBP, 3β-HSD and CCND1 were promoted, and PPARγ, CYP19, BCL2 and CAS3 were inhibited (p < .05); in the hierarchical TCs, the expression levels of PPARγ, FAS, CYP19, CCND1 and BCL2 were promoted, and SREBP, STAR, 3β-HSD and CAS3 were inhibited (p < .05), whereas in the F1 TCs, the expression levels of PPARγ, FAS, 3β-HSD, CYP19 and CCND1 were promoted, and STAR and CAS3 were inhibited (p < .05). These results suggested that GCs at the three FDSs have dynamic and complex influences on the physiological characteristics of TCs, and the influences on TCs at the three FDSs were varied.     

Characterization of Necrotoxigenic Escherichia coli (NTEC) Strains Isolated from Healthy Calves in Poland

Faecal samples from 132 healthy, 4–8‐week‐old calves from four different farms were examined for necrotoxigenic Escherichia coli (NTEC) producing the cytotoxic necrotizing factors type 1 (CNF1) and type 2 (CNF2). CNF2 genes were detected by polymerase chain reaction in 24 (6.1%) of the 396 E. coli strains tested; these strains were found in 18 (13.6%) calves used in the study. None of the 396 E. coli isolates examined possessed the gene encoding CNF1. Overall, 28.8% of E. coli examined expressed the F17 fimbrial antigen. A strong association between CNF2 toxin and F17 fimbriae was found (62.5% of CNF2‐positive strains were F17‐positive). Moreover, six out of 24 NTEC strains had the Stx1 or the Stx2 shiga toxin genes, and three additional isolates possessed the eae genetic marker of the intimin protein.