Chicken PRDX3 is required for proliferation of chicken embryo fibroblast cells

ABSTRACT

1. This experiment investigated the influence of chicken PRDX3 on cell proliferation in chick embryo fibroblast cells using PRDX3 knockdown technology.

2. A methyl thiazolyl tetrazolium (MTT) assay was performed to assess the effect of chPRDX3 knockdown on fibroblast proliferation. The antioxidant effect was investigated to determine if it directly mediated fibroblast cell proliferation.

3. To determine the role of chPRDX3 on cell proliferation, an siRNA mediated knockdown was performed in chick fibroblast cells using an in vitro assay. The proliferation of fibroblast cells transfected with siPRDX3 #3 and siPRDX3 Mix was significantly decreased after 48 h (P < 0.01). In addition, the knockdown of chicken PRDX3 suppressed cell proliferation through an increase in oxidative stress.

4. The results demonstrated that chPRDX3 is required for cell proliferation in chicken fibroblast cells. Such findings have important implications for the maintenance of chicken fibroblast cells.     

Interactions between genes involved in growth and muscularity in pigs: IGF-2, myostatin, ryanodine receptor 1, and melanocortin-4 receptor

In the swine breeding industry, two economical traits are of particular importance in sires, namely, muscle growth and average daily gain (ADG). These traits are quantitative, which implies that they are under the control of multiple genes. Mutations in these genes, associated with either muscularity or growth, are useful quantitative trait nucleotides (QTN) for unraveling genetic variation of these traits and can be used in marker-assisted selection. Until now, QTN involved in muscle growth and/or ADG in pigs were identified in porcine ryanodine receptor 1 (RYR1), insulin-like growth factor-2 (IGF-2), and melanocortin-4 receptor (MC4R). Recently, a fourth possible QTN was found in porcine myostatin (MSTN). All four QTN have an influence on muscle growth and/or somatic growth, so an influence of one mutation on one or more of the other mutations should not be excluded. However, although the polymorphisms in the RYR1 and the MC4R gene affect the function of the respective protein, the polymorphisms of the IGF-2 and MSTN gene influence the mRNA expression of the respective gene. Therefore, this study investigated possible interactions between the genotypes of MSTN, IGF-2, and MC4R (population 1) or the RYR1, IGF-2, and MSTN QTN (population 2) on IGF-2 and MSTN expression in different muscle types in pigs. In both skeletal muscle and heart muscle growth, the IGF-2:MSTN ratio seems to play an important role. Also, the RYR1 genotype had a significant effect on IGF-2 expression in m. longissimus dorsi. No effect of the MC4R QTN could be seen.     

The Use of Tannins to Control Salmonella Typhimurium Infections in Pigs

The aim of this study was to determine whether a hydrolysable tannin extract of sweet chestnut wood (Globatan^®) has an inhibitory effect on Salmonella Typhimurium survival both in vitro and in vivo in pigs. In a first experiment, the minimal inhibitory concentration of Globatan^® on 57 Salmonella Typhimurium isolates was determined. For all isolates, an MIC of 160–320 μg/ml was found. The second in vitro study revealed that Salmonella growth was strongly reduced using Globatan^® concentrations of 25–50 μg/ml and nearly completely inhibited at a concentration of 100 μg/ml Globatan^®. In an in vivo trial, two groups of six piglets, each group receiving feed with or without the addition of Globatan^® (3 g/kg), were orally inoculated with 10⁷ colony forming units of a Salmonella Typhimurium strain. Globatan^® had no effect on faecal excretion of Salmonella, and no differences in colonization of the intestines and internal organs were demonstrated in pigs euthanized at 4 days post‐inoculation. In conclusion, the hydrolysable tannin extract used in this study showed strong action against Salmonella Typhimurium in vitro but not in vivo.     

Studies on the Parietal Region of the Cervid Skull III. On the Occurrence of an Interparietal in Cervus

The occurrence of an os interparietale was studied in two transparent preparations of fetal red deer (Cervus elaphus) heads and in 14 dried skulls of fetal to early postnatal individuals from four Cervus species (C. elaphus, C. nippon, C. duvauceli and C. eldi). In 14 of the 16 specimens, an interparietal was present as either a paired or single bone. In only a neonate red deer and a 5-week-old sika deer this skull element was missing. We therefore conclude that an os interparietale, developing from paired centres of ossification, is normally present in Cervus species. This clearly distinguishes them from the fallow deer, where an interparietal is missing (Kierdorf and Kierdorf, 1992b). Our findings thus support the view that the fallow deer should be considered a distinct genus Dama instead of being included within Cervus.     

First account of the metazoan parasite fauna of oilfish Ruvettus pretiosus Cocco, 1829 (Perciformes: Gempylidae) in South African waters

Parasites are an important but neglected component of ecosystems that can be used as indicators of host biology and ecology. In the present study, the metazoan parasite assemblage of Ruvettus pretiosus, an understudied but widely distributed predatory gempylid, caught off South Africa was surveyed. A total of seven parasite taxa, including four new infection records (Bolbosoma capitatum, Rhadinorhynchus sp., Hepatoxylon trichiuri and Anisakis sp.), two new locality records (B. capitatum and Rhipidocotyle sp.) as well as the ectoparasitic copepod Sagum foliaceus and the cestode Tentacularia coryphaenae were recovered from the eight specimens examined.     

Preliminary molecular analysis of Clostridium difficile isolates from healthy horses in northern Italy

Clostridium difficile, associated with a wide spectrum of diseases in humans, as well as in several animal species, is an important cause of colitis in adult horses and foals. The aim of this study was to investigate by toxin gene profile and PCR-ribotyping the molecular characteristics of 14 C. difficile strains isolated from 42 faeces of healthy horses. Both toxin genes, tcdA and tcdB, were present in only 1 isolate (7.1%). Six isolates (42.9%) demonstrated tcdA−/tcdB+ genotype, and seven isolates (50.0%) were tcdA−/tcdB−. All strains were binary toxin genes negative (cdtA−/cdtB−). The PCR-positive strains, except for the tcdA+/tcdB+ isolate, tested negative for, in vitro, A and/or B toxins production by EIA. Eleven distinct ribotypes were observed.In conclusion, C. difficile can be present in the normal intestinal flora of healthy adult horses, in addition to foals. These animals could therefore play an important role as potential reservoirs of toxigenic strains.     

Pattern of Eimeria Oocyst Output and Repeatability in Naturally Infected Suckling Rhön Lambs

The faeces of 14 Rhön lambs were examined every second day between 7 and 99 days of age for the presence of Eimeria oocysts. Eimeria ahsata, E. bakuensis, E. faurei, E. granulosa, E. intricata, E.ovinoidalis, E. pallida, E. parva and E. crandallis/weybridgensis were identified. The predominant species were E. ovinoidalis, E. parva, E. crandallis/weybridgensis and E. bakuensis. Using a statistical model, the oocyst excretion rate was described as a sequence of periods with decreasing levels and varying length (‘excretion periods’) interrupted by intervals with no or very low oocyst counts. Several variables could be deduced from these two parameters, including the length of an excretion period and the maximum output during an excretion period. The estimated repeatability for oocyst counts for the different species ranged from 0.05 to 0.41. This result provides a starting point for possible genetic selection based on faecal oocyst counts of Rhön sheep for resistance to Eimeria infections.     

Association of BoLA‐DRB3 alleles with mastitis resistance and susceptibility in Japanese Holstein cows

In this study, 714 cows from 26 dairy herds were reclassified as healthy or mastitic cows on the basis of long‐term somatic cell count (SCC) in milk. Cows with more than three consecutive lactation records of SCC from the first or second to fifth lactation, were selected, and their BoLA‐DRB3 (DRB3) alleles were identified using polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) method. Cows with an SCC of < 200 000 cells/mL in all monthly records were classified as healthy (n = 91). Cows with an SCC of > 300 000 cells/mL in two consecutive tests or four non‐consecutive tests or cows with an SCC of > 500 000 cells/mL in any one test during lactation, regardless of parity, were classified as mastitic (n = 201). Mastitic cows (n = 153) from another 40 herds were considered to be infected if bacteriological testing revealed mastitis pathogens in milk. Their DRB3 alleles were identified using PCR‐sequence‐based typing (PCR‐SBT). The differences in DRB3 allelic frequencies between healthy cows and cows with various degrees of mastitis were re‐investigated. Moreover, the associations of various amino acid motifs in DRB3 alleles with resistance or susceptibility to mastitis pathogens were re‐examined. DRB3.2*8(DRB3*1201) and DRB3.2*16(DRB3*1501) alleles were found to be associated with susceptibility, while DRB3.2*22(DRB3*1101), DRB3.2*23(DRB3*2703), and DRB3.2*24(DRB3*0101) alleles were found to be associated with resistance.     

An enclosure study to evaluate the effect of Sargochromis codringtoni on snail populations in an irrigation canal

The cichlid fish, Sargochromis codringtoni has been suggested as an alternative to the use of molluscicides for controlling schistosome intermediate host snails. To determine the effect of the fish on Bulinus globosus and Biomphalaria pfeifferi populations in irrigation canals, an enclosure experiment was carried out. The presence of fish was found to significantly reduce the numbers of both B. globosus and B. pfeifferi.     

凝结芽孢杆菌对感染产气荚膜梭菌肉鸡生长性能、肠道病变及免疫器官指数的影响

本试验旨在研究日粮中添加凝结芽孢杆菌对感染产气荚膜梭菌肉鸡生长性能、肠道病变及免疫器官指数的影响。选用312只1日龄科宝500肉鸡公雏,随机分成4个处理,每个处理6个重复,每个重复13只鸡。采用2×2双因子完全随机设计,产气荚膜梭菌分为无感染产气荚膜梭菌组和感染产气荚膜梭菌组,日粮中凝结芽孢杆菌水平分别为0和400mg/kg。试验期35d。结果表明,感染产气荚膜梭菌后15~35日龄肉鸡平均日增重显著降低(P0.05),28日龄肠道病变严重(P0.01),35日龄法氏囊指数极显著降低(P0.01);日粮中添加凝结芽孢杆菌可显著提高15~35日龄肉鸡平均日增重(P0.05),显著降低料重比(P0.05),能缓解感染导致的肠道损伤(P0.05),28日龄肉鸡胸腺指数极显著提高(P0.01),35日龄法氏囊指数极显著提高(P0.01)。肠道病变结果分析表明,日粮中添加凝结芽孢杆菌与感染产气荚膜梭菌有显著交互作用(P0.05)。综上所述,日粮中添加凝结芽孢杆菌能够提高感染产气荚膜梭菌肉鸡生长性能及免疫器官指数,增强动物免疫能力,并能缓解由于产气荚膜梭菌感染而导致的不利影响。     

Genetic variation in the ovine uncoupling protein 1 gene: association with carcass traits in New Zealand (NZ) Romney sheep,but no association with growth traits in either NZ Romney or NZ Suffolk sheep

The uncoupling protein 1 (UCP1) plays an important role in the regulation of lipolysis and thermogenesis in adipose tissues. Genetic variation within three regions (the promoter, intron 2 and exon 5) of the ovine UCP1 gene (UCP1) was investigated using polymerase chain reaction‐single‐strand conformational polymorphism (PCR‐SSCP) analyses. These revealed three promoter variants (designated A, B and C) and two intron 2 variants (a and b). The association of this genetic variation with variation in lamb carcass traits and postweaning growth was investigated in New Zealand (NZ) Romney and Suffolk sheep. The presence of B in a lamb's genotype was associated with decreased subcutaneous carcass fat depth (V‐GR) (p = 0.004) and proportion of total lean meat yield of loin meat (p = 0.005), and an increased proportion of total lean meat yield of hind‐leg meat (p = 0.018). In contrast, having two copies of C was associated with increased V‐GR (p < 0.001) and proportion of total lean meat yield of shoulder meat (p = 0.009), and a decreased hind‐leg yield (p = 0.032). No associations were found with postweaning growth. These results suggest that ovine UCP1 is a potential gene marker for carcass traits.     

Vegetative Bacillus amyloliquefaciens cells do not confer protection against necrotic enteritis in broilers despite high antibacterial activity of its supernatant against Clostridium perfringens in vitro

1. In this study, the effect of Bacillus amyloliquefaciens on Clostridium perfringens was tested in vitro and in vivo.

2. Using an agar well diffusion assay, the inhibitory activity of B. amyloliquefaciens supernatant was analysed against a large collection of netB-positive and netB-negative C. perfringens strains. Although strong growth inhibiting activity was detected against all C. perfringens isolates, it was significantly higher against virulent netB-positive C. perfringens strains compared with avirulent netB-negative isolates.

3. Subsequently, the efficacy of in-feed administration of lyophilised vegetative cells of B. amyloliquefaciens to prevent necrotic enteritis was tested in vivo using an established experimental infection model in broilers. Ross 308 broilers received either B. amyloliquefaciens supplemented or unsupplemented feed throughout the experiment. No significant differences could be detected between the untreated positive control group and the B. amyloliquefaciens treated group in body weight, the number of chickens that developed necrotic lesions and in pathological lesion scores.

4. These results demonstrate that despite its substantial inhibitory activity in vitro, lyophilised vegetative B. amyloliquefaciens cells had no beneficial effect against necrotic enteritis in the in vivo model used here.

     

Life-table experiments with Helisoma duryi (Wetherby) and Biomphalaria Pfeifferi (Krauss) at constant temperatures

In an effort to find an ecological homologue that could be utilized as a biological control agent of the intermediate host species of economically important helminth parasites, age-specific life-table experiments with Helisoma duryi (Wetherby) ind Biomphalaria pfeifferi (Krauss) (intermediate host of Schistosoma mansoni) were carried out at a series of constant temperatures in a laboratory. The results showed that the lifespan and net reproduction rate of H. duryi were higher than those of B. pfeifferi over the whole range of temperatures investigated (18-35°C). The conclusion is reached that H. duryi should theoretically be capable of establishing viable populations in most of the habitats occupied by B. pfeifferi in South Africa.     

Detection and Identification of Fish-Pathogenic Aphanomyces piscicida Using Polymerase Chain Reaction (PCR) with Species-Specific Primers

Abstract

Aphanomyces piscicida is an important pathogen that plays a role in the morbidity and mortality of various fish species around the world. The poor quality of current identification techniques for this fungus led to the development of highly sensitive and specific polymerase chain reaction (PCR) techniques. Primers derived from the ITS1 and ITS2 regions of A. piscicida NJM 0204 were designed for the detection and identification of fish-pathogenic A. piscicida, with the potential for the diagnosis of mycotic granulomatosis (MG). Polymerase chain reaction amplification showed that the primer set was specific only to fish-pathogenic A. piscicida, not to non-fish-pathogenic Aphanomyces, Saprolegnia spp., Achlya spp., Dictyuchus spp., and Lagenidium spp. In addition, PCR revealed an improved sensitivity sufficient to detect A. piscicida in artificially infected goldfish Carassius auratus. Results demonstrated that the PCR method established in this study is effective for the detection and identification of A. piscicida with MG.     

Detection and genotypic differentiation of Campylobacter jejuni and Campylobacter coli strains from laying hens by multiplex PCR and fla-typing

In total, 26 Campylobacter (C.) strains, isolated from liver, spleen, caecal or jejunal content of laying hens from different flocks were examined. In these flocks a drop in egg production, an increasing mortality and livers with whitish-grey lesions as post-mortem finding were observed. Suspected Campylobacter colonies were differentiated using a modified m-PCR in 13 Campylobacter jejuni and 13 Campylobacter coli strains. All isolates were characterised by typing of the flaA and flaB gene each with two restriction enzymes. To compare the four different profiles for all strains an artificial “fla-type” was generated. Different and identical fla-types of C. jejuni and C. coli were recovered from both intestinal and extra-intestinal organs of the laying hens and even from individual birds. One significant observation is that some fla-types of C. jejuni or C. coli were detected in intestinal and systemic sites but not all fla-types of both species appeared to be equally able to invade internal organs.     

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.     

Clinical Leptospira interrogans serogroup Australis serovar lora infection in a stud farm in the Netherlands

Summary

A Leptospira interrogans serogroup australis serovar lora infection in a stud farm is reported. During three successive years (1984–1986) clinical leptospirosis with a severe often rapid, fatal course was seen in 12 foals.

Clinical examination revealed severe respiratory distress, depression and pyrexia. Other symptoms were diarrhea (2), jaundice (1), and an unsteady gait (1). Morphological characteristics of the disease were massive pulmonary haemorrhage and haemorrhagicthrombotic or extracapillary glomerulonephritis with tubulonephrosis and interstitial oedema. In most foals high or increasing MAT titres to serovar bratislava were found; from one foal Leptospira interrogans serovar lora was isolated.

Serological examination of all 56 mares at the farm (August 1986) revealed antibodies to serovar bratislava in 64 per cent of the animals. These findings support the idea that Leptospira interrogans serovar bratislava and closely related strains (in this study serovar lora) may be adapted to and maintained by the horse population.     

Detection of Myxobolus (Myxosoma) cerebralis in Salmonid Fishes in Oregon

Abstract

Myxobolus (Myxosoma) cerebralis, the etiological agent of whirling disease, was detected in salmonid fish populations in northeastern Oregon. This is the first record of M. cerebralis in the Pacific Northwest of the USA. During an epizootiological survey for the parasite, two methods for spore detection were compared, and an efficient procedure for determining M. cerebralis infection in adult fish was developed. The enzyme digest method was more efficient than the plankton centrifuge procedure for examination of numerous individual lots of fish processed during the survey. Sampling only the area around the otoliths was at least as effective as sampling entire heads for detection of spores in infected fish.     

Dry Matter and Crude Protein Degradability of Four Parasitic Plants (Mistletoes) Associated with Browse Trees in Botswana

Four parasitic plants (Tapinanthus lugardii, Erianthenum ngamicum, Viscum rotundifolium and Viscum verrucosum) associated with browse trees in Botswana were evaluated for the degradability of their dry matter (DM) and crude protein (CP). All these plants have high levels of crude protein, ranging from 144.5 to 163.4 g/kg DM, with a significant (p<0.001) difference in crude protein content between the plants. The rate of degradation of DM was highest for V. rotundifolium (0.197 h⁻¹) and lowest for E. ngamicum (0.031 h⁻¹). The two Viscum species had the highest effective DM degradability at an outflow rate of 0.05%. The rate of degradation of crude protein was highest for V. rotundifolium (0.144 h⁻¹) and lowest for T. lugardii (0.0346 h⁻¹). The effective degradability (ED) of crude protein was highest for V. rotundifolium at 0.03 and 0.05 h⁻¹ outflow rates. T. lugardii and E. ngamicum had an apparently low effective degradability, but this may result from an association of their protein with tannins, which would be able to supply amino acids in the small intestine after dissociation due to the abomasal pH. This revised version was published online in July 2006 with corrections to the Cover Date.