Diagnosis of sub-clinical coccidiosis in fast growing broiler chickens by MicroRNA profiling

Coccidiosis in broiler chickens, caused by infection with Eimeria spp. remains one of the most economically important production diseases. Development of a genetic biomarker panel of sub-clinical infection would be an important biological tool for the management of broiler flocks.We analysed expression of MicroRNAs (miRNAs) to determine the potential for these in diagnosing coccidiosis in broiler flocks. miRNA expression, in the ilea of Ross 308 broilers, was compared between chickens naturally clinically or sub-clinically infected with Eimeria maxima and Eimeria acervulina using NextSeq 500 sequencing. 50 miRNAs with greatest coefficient of variance were determined and principal component analysis showed that these miRNAs clustered within the clinical and sub-clinical groups much more closely than uninfected controls. Following false detection rate analysis and quantitative PCR we validated 3 miRNAs; Gallus gallus (gga)-miR-122-5p, gga-miR-205b and gga-miR-144-3p, which may be used to diagnose sub-clinical coccidiosis.     

Spontaneous hypoxaemia and right ventricular hypertrophy in fast growing broiler chickens reared at sea level

1. At 6 weeks of age, the time of most rapid body growth, fast growing broiler chickens showed more right ventricular hypertrophy than slower growing chickens. 2. The degree of right ventricular hypertrophy was directly related to blood haematocrit and indirectly related to arterial oxygen saturation (estimated in the chickens using an ear oximeter designed for man). 3. When oxygen saturation was estimated sequentially from 6 to 17 weeks of age, mean oxygenation improved with age, partly due to the death of the chickens with the worst saturations, but also because of an improvement in oxygenation of the survivors.     

Differentiation of slow and fast muscles in chickens

1. The development of the characteristic histochemical appearance of the slow anterior latissimus dorsi (ALD) and fast posterior latissimus dorsi (PLD) was studied in chickens during embryonic development as well as during regeneration of minced muscle. 2. During embryonic development the activity of the oxidative enzyme succinic dehydrogenase (SDH) is higher in the slow ALD muscle already at 16 days of incubation. At this time the fast PLD has a higher activity of the glycolytic enzyme, phosphorylase. Although the histochemical appearance of the two types of muscle is already different at 16 days, their contractile speeds are still similar. No difference in myosin ATP-ase was found in the two muscles in young embryos but in 20-day old embryos the two muscles became distinctly different when stained for this enzyme. 3. When PLD muscles in hatched chickens redeveloped during regeneration in place of ALD the histochemical characteristics of the regenerated muscle resembled ALD, and when ALD regenerated in place of PLD it resembled PLD. 4. It is concluded that the histochemical characteristics of slow and fast muscles become determined during early development, even before any difference in contractile properties can be detected and that they are determined by the nerve.     

Behavioural effects of embryonic exposure to corticosterone in chickens

We tested the hypothesis that exposure of chick embryos to corticosterone leads to increased fear, reduced competitive ability, reduced ability to cross a barrier and reduced growth in juvenile chicks. Behaviour was studied in birds subjected to three different egg injection treatments: a negative control (no treatment of eggs), a positive control (100 μl sesame oil vehicle) and a corticosterone treatment (0.6 μg corticosterone in 100 μl sesame oil). Eggs were injected prior to incubation and the behaviour of chicks was studied during the first 4 weeks of life. Corticosterone treatment increased fear in chicks, as indicated by greater avoidance of an observer in the home pen at 2 weeks of age (P < 0.0001), reduced ability to cross a wall to access feed at 2 weeks of age (P < 0.05) and reduced ability to compete for a wormlike object at 4 weeks of age (P < 0.01). Treatment with corticosterone also reduced body weight at 1 week of age (P < 0.003) and 4 weeks of age (P < 0.04), but not at hatch (P < 0.28). The sesame oil vehicle reduced fear (P < 0.0001), but had no other significant effects. These results indicate that embryonic exposure to corticosterone leads to behavioural and growth deficits in chicks.     

Body weight and metabolic level in growing chickens

Body weight and metabolic level in growing chickens. Acta Physiol. Pol., 1977, 28 (6): 575-583. In the investigations carried out on 128 White Rock chicken the metabolic level was determined under standard conditions in chickens characterising by different growth rate resulted from different feeding levels. It was shown that the feeding level of growing chicken has an essential influence on the character of the regression between the metabolic level and body weight. It was also found that this regression is not necessarily ot the character of Kleiber's function H = aWb. The differences in the metabolic level between the chickens maintained on different feeding levels did not disappear even after 4 days of starvation. In the light of the obtained results and recent investigations by other authors the usefulness of the so-called metabolic unit of the body size based on W0.75 in context to growing animals within one species has bee criticized.     

Heat production and lipid metabolism in broiler and layer chickens during embryonic development

We compared heat production (HP) and lipid metabolism in broiler and layer chickens (Gallus gallus) during embryonic development. To investigate HP and respiratory quotient (RQ), oxygen (O2) consumption and carbon dioxide (CO2) production were measured using an open-circuit calorimeter system. HP consistently had a tendency (P = 0.06) to be lower in broilers than in layers during embryonic development, and HP gradually decreased with developmental stage in both strains. RQ values of both strains were approximately 0.7 at every embryonic stage investigated. These results suggest that chicken embryos mainly use lipid for energy, and the RQ was significantly lower in broilers than in layers during embryonic development. Consumption of the yolk sac as a lipid source was faster in broilers than in layers. Plasma D-3-hydroxybutyrate (D3HB) and glycerol concentrations, associated with fatty acid oxidation, were lower in broiler than layer embryos. These results demonstrate that HP and lipid metabolism are different between the strains during embryonic development, and may be one factor for the growth difference between broiler and layer embryos.     

Differences of Z chromosome and genomic expression between early- and late-feathering chickens

One duplicated segment on chicken Z chromosome is a causal mutation to the late-feathering phenotype. However, understanding biological process of the late-feathering formation is also of interest to chicken breeding and feather development theory. One hundred and thirty-seven valid single nucleotide polymorphisms (SNPs) from an SNP database were used to perform an association study of the Z chromosome in Xinghua chickens. Two SNPs, which were respectively on 9607480 bp and 10607757 bp, were significantly associated with feathering phenotypes. This result indicated the causal mutation of the late-feathering formation in Xinghua chickens was consistent with the previous report which showed the late-feathering locus ranged 9966364–10142688 bp on Z chromosome. Microarray expressions were implemented for six 1-day-old female Xinghua chicks. Compared to the early-feathering chicks, there were 249 and 83 upregulated and downregulated known genes in the late-feathering chicks. Forty-one genes were expressed in late-feathering chicks, but not in early-feathering ones. At least 14 significantly differentially expressed genes were directly related to keratin. In the region of the sex-linked feathering gene, only prolactin receptor (PRLR) gene was a significantly differentially expressed gene. Expression of PRLR in late-feathering chicks was 1.78-fold as that in early-feathering chicks. Late-feathering Wenchang chicks also had higher expression level of PRLR than early-feathering ones. This study suggested that increasing PRLR expression that resulted from the special variant on chicken Z chromosome caused the late-feathering phenotype.     

Comparative study of myocardial high energy phosphate substrate content in slow and fast growing chicken and in chickens with heart failure and ascites

In order to explain the biochemical mechanisms associated with deteriorating heart function in broiler chickens, this study compared myocardial high energy phosphate substrates in leghorns, feed restricted (Broilers-Res) broilers, ad libitum fed broilers (Broilers-AL), and in broilers that developed heart failure and ascites. The profile of adenine nucleotide content in the heart tissue did not differ between leghorns and Broilers-Res, but there were significant differences among Broilers-Res, Broilers-AL, and broilers with ascites. During intensive growth periods, leghorns and Broilers-Res showed increasing trends in heart ATP levels, whereas in fast growing broilers the heart ATP declined (p<0.021). ATP:ADP and ATP:CrP ratios increased with age in both leghorn and Broilers-Res, declined in fast growing broilers, and were the lowest in broilers that developed heart failure. The changes in heart high energy phosphate profile in broilers suggest that the energy demand of the heart during a rapid growth phase may exceed the bird's metabolic capacity to supply adequate levels of high energy phosphate substrate. The insufficiency of energy substrate likely contributes to the declining heart rate. In some individuals this may lead to impaired heart pump function, and in more severe cases may progress to heart pump failure.     

Effect of Al and heavy metals on enzymes of nitrogen metabolism of fast and slow growing rhizobia under explanta conditions

Most of the legume crops are affected by metal stress present in the soil mainly due to contaminated agrochemicals and sewage sludge. The effect of aluminium, and heavy metals copper, iron and molybdenum on growth and activity of enzymes of fast and slow growing rhizobial sps. was studied. Sinorhizobium meliloti RMP₅ was found to be more tolerant to metal stress than Bradyrhizobium BMP₁. Both the strains were extremely sensitive to Al than other metals. Al was much more deleterious for the enzymatic activities (nitrate reduction, nitrite reduction, nitrogenase and uptake hydrogenase) of strain RMP₅ and BMP₁. Cu showed inhibitory effect on growth and enzyme activities of Bradyrhizobium strain at all concentrations. However, in S. meliloti RMP₅ all the tested enzymatic activities increased up to the concentration of 0.1 mM Cu. Fe enhanced the growth and enzyme activities of S. meliloti RMP₅ and Bradyrhizobium BMP₁ up to 100 mM concentration. Mo enhanced all the tested enzymatic activities of S. meliloti RMP₅ up to 1 mM. Nitrate and nitrite reduction activities of Bradyrhizobium BMP₁ increased up to 1 mM concentration. However, nitrogenase and hydrogenase activities of Bradyrhizobium BMP₁ got enhanced only up to 0.5 mM Mo. Both Fe and Mo are the key components of the enzyme nitrogenase and nitrate reductase and enhanced the growth and enzyme activities of both the sps. The study of physiology of nitrogen fixing ability of both fast and slow growing rhizobial strains reported that the supplementation of Mo and Fe in soils along with the biological formulations will enhance the process of symbiotic nitrogen fixation.     

Indigenous bacteria and bacterial metabolic products in the gastrointestinal tract of broiler chickens

The gastrointestinal tract is a dynamic ecosystem containing a complex microbial community. In this paper, the indigenous intestinal bacteria and the microbial fermentation profile particularly short chain fatty acids (SCFA), lactate, and ammonia concentrations are reviewed. The intestinal bacterial composition changes with age. The bacterial density of the small intestine increases with age and comprises of lactobacilli, streptococci, enterobacteria, fusobacteria and eubacteria. Strict anaerobes (anaerobic gram-positive cocci, Eubacterium spp., Clostridium spp., Lactobacillus spp., Fusobacterium spp. and Bacteroides) are predominating caecal bacteria in young broilers. Data from culture-based studies showed that bifidobacteria could not be isolated from young birds, but were recovered from four-week-old broilers. Caecal lactobacilli accounted for 1.5-24% of the caecal bacteria. Gene sequencing of caecal DNA extracts showed that the majority of bacteria belonged to Clostridiaceae. Intestinal bacterial community is influenced by the dietary ingredients, nutrient levels and physical structure of feed. SCFA and other metabolic products are affected by diet formulation and age. Additional studies are required to know the bacterial metabolic activities together with the community analysis of the intestinal bacteria. Feed composition and processing have great potential to influence the activities of intestinal bacteria towards a desired direction in order to support animal health, well-being and microbial safety of broiler meat.     

Analysis of metabolic systems with complex slow and fast dynamics

We briefly review the results of other authors concerning the analysis of systems with time hierarchy, especially the Tikhonov theorem. A theorem, recently proved by the authors, making possible rigorous analysis of systems with complex fast dynamics is stated and discussed. A model example of a simple enzymatic reaction with product activation and slow (genetically driven) enzyme turnover is rigorously studied. It is shown that even in such a simple model there exist certain regions of parameters for which fast variables oscillate. Thus the classical Tikhonov theorem is not applicable here and we are forced to use another method-for example the author's presented theorem—or a purely numerical solution. These two methods are compared.     

Effect of corticosterone on growth and welfare of broiler chickens showing long or short tonic immobility

Tonic immobility (TI) test is commonly used to assess fear. Animals showing different TI durations demonstrate distinct behavior and biochemical responses to stress. However, less is known about how TI phenotype affects growth and welfare of domestic fowl. In this study, broiler chickens (Gallus gallus) were classified into short and long TI duration (STI and LTI) phenotypes and treated chronically with vehicle (CON) or corticosterone (CORT). STI broilers demonstrated significantly higher growth rate with higher breast muscle yield (P < 0.05) and liver weight relative to BW tended to be lower (P = 0.053), which was accompanied by higher serum concentration of CORT (P < 0.05) and uric acid (P < 0.01), but lower serum level of T4 (P = 0.01). CORT severely reduced body weight, as well as the relative weight of muscle, bursa of Fabricius and spleen (P < 0.001), but relative liver weight was increased (P < 0.001). CORT-treated chickens had reduced serum CORT, elevated heterophile/lymphocyte ratio, and increased serum levels of total and free T3. STI broilers displayed more preening behavior (P < 0.05), yet CORT elicited more walking behavior (P < 0.05). No difference was observed in the welfare assessment scores between STI and LTI phenotypes under basal situation, while LTI chickens showed significantly increased incidence of pad dermatitis compared to STI under CORT exposure. The results suggest that STI broilers demonstrate better growth performance and higher adaptability to stress compared to LTI chickens.     

Comparative study of the DNA-binding HU-type proteins from slow growing and fast growing strains of Rhizobiaceae

The DNA-binding HU-type proteins have been isolated from two very different strains of Rhizobiaceae: Agrobacterium tumefaciens and Rhizobium japonicum. These proteins have been called HAt and HRj respectively. Their electrophoretic mobility on polyacrylamide gel, amino acid composition and crossed immunoreactivity have been compared to that of the homologous protein isolated from Rhizobium meliloti: the protein HRm. The proteins HAt and HRm show close similarities whereas the protein HRj differs markedly from the two others. The physico-chemical characteristics of the HU-type proteins from these Rhizobiaceae are in good agreement with the respective position of these bacteria in the taxonomy.     

Structure of the egg funiculus and deposition of embryonic envelopes in a crab

The newly laid egg of Carcinus maenas is attached to a maternal ovigerous seta by a funiculus which consists of the two superimposed vitelline envelopes 1a + 1b, highly stretched and concurrently showing important structural alterations. The funiculus is glued to the specialized seta merely owing to the strong adhesiveness of its external face comprising the outermost vitelline envelope 1a, without any added adhesive. The subjacent envelope 2, originated from the cortical reaction, is not involved in such a funiculus elaboration. In the course of the embryonic development, four new coatings are successively secreted from the ectodermal embryonic cells, underneath the (1a + 1b + 2) fertilization envelope or embryonic capsule. They will remain until hatching in this concentric order, thus giving evidence of successive embryonic moulting cycles, with apolysis but without exuviation. In addition, the successive secretory phases, regarding to the embryonic envelope elaborations, happen in presence of high concentrations of the ecdysteroid ponasterone A which might be involved consequently in such secretory processes.     

IGF-I and IGF-II mRNA expression in slow and fast growing families of USDA103 channel catfish (Ictalurus punctatus)

The objective of this study was to examine insulin-like growth factor (IGF)-I and IGF-II mRNA levels in fast and slow growing families of catfish. Relative levels of IGF-I and IGF-II mRNA were determined by real-time PCR. Family A exhibited a specific growth rate (SGR) of 3.6 and was designated as fast growing, while family H exhibited a SGR of 3.1 and was designated as slow growing (P=0.017). Levels of IGF-II mRNA were 3.3-fold greater (P=0.006) in muscle for the fast growing family compared to the slow growing family. Levels of IGF-II mRNA were 1.8-fold greater (P=0.049) in liver for the fast growing family compared to the slow growing family. Levels of IGF-II mRNA from both fast and slow families were 12.2-fold greater (P<0.001) in muscle and 5.8-fold greater (P=0.021) in liver, respectively, compared to levels of IGF-I mRNA. Muscle and liver levels of IGF-I mRNA were similar between families. Elevated levels of IGF-II mRNA in muscle and liver compared to IGF-I mRNA, as well as differences in levels of IGF-II mRNA between fast and slow growing families of fish suggests a role of IGF-II in growth of channel catfish.     

Altered development and protein metabolism in skeletal muscles of broiler chickens (Gallus gallus domesticus) by corticosterone

Two trials were conducted to investigate the effect of corticosterone (CORT) on protein metabolism and the amino acid composition in muscle tissues of broiler chickens (Gallus gallus domesticus). In Trial 1, two groups of 30 broiler chickens were subjected to control or CORT treatment (30 mg/kg diet) from 28 to 39 days of age. In Trial 2, three groups of chickens of 28 days of age were randomly subjected to one of the following treatments for 7 days: CORT (30 mg/kg diet), pair-fed (maintaining the same feed intake as CORT treatment) and control treatments. The body mass gain and feed efficiency was significantly decreased by CORT treatment, while the food intake was decreased. The breast and thigh masses (% body mass) were significantly suppressed by CORT treatment, while the abdominal fat and liver masses (%) were obviously increased. The plasma levels of glucose, urate and total amino acid were significantly elevated by CORT treatment. The capacity for protein synthesis, estimated by RNA:protein ratio, were significantly suppressed by CORT in M. pectoralis major and M. biceps femoris. The 3-methylhistidine concentrations were significantly increased in both M. pectoralis major and M. biceps femoris of CORT chickens, compared to control but not the pair-fed chickens. The amino acid composition of M. pectoralis major and M. biceps femoris was not significantly affected by CORT treatment. In conclusion, the arrested growth in skeletal muscles induced by CORT administration has tissue specificity. The CORT treatment retards the growth of skeletal muscle by suppressed protein synthesis and augmented protein catabolism.     

Changes in free amino acids in the brain during embryonic development in layer and broiler chickens

Developmental changes in the levels of the excitatory amino acids l-glutamate (Glu) and l-Aspartate (Asp) and inhibitory amino acids glycine (Gly) and γ-amino butyric acid (GABA), as well as taurine and its related amino acids l-methionine (Met), l-cysteine (Cys) and l-serine (Ser) in the brain and pectoralis muscle at various embryonic stages and hatch in broiler and layer type chickens were determined. Brain concentrations of Asp, GABA and taurine were higher than those in the muscle, but the difference in the two types was small. The concentrations of the precursors of taurine including Met, Cys and Ser were lower than that of taurine. In conclusion, the synthesis of some amino acids and their metabolites such as Asp, GABA and taurine in the chick embryo is very high in order to support brain development.