Effect of epidermal growth factor and insulin-like growth factor I on porcine preantral follicular growth, antrum formation, and stimulation of granulosal cell proliferation and suppression of apoptosis in vitro

The object of this study was to investigate the role of epidermal growth factor (EGF) and IGF-I in the regulation of preantral follicular growth, antrum formation, and granulosal cell proliferation/ apoptosis. Porcine preantral follicles were manually dissected and cultured for up to 8 d in Waymouth's (Exp. 1) or alpha-minimum Eagle's essential medium (Exp. 2 and 3) supplemented with 10 microg/mL of transferrin, 100 microg/mL of L-ascorbic acid, and 2 mU/mL of ovine FSH, in the presence (Exp. 1 and 3) or absence (Exp. 2) of 7.5% fetal calf serum. According to the experimental protocol, IGF-I (0, 1, 10, or 100 ng/mL; Exp. 1), or IGF-I (50 ng/mL), EGF (10 ng/mL) and EGF+IGF-I (Exp. 2 and 3) were added to the culture media. In Exp. 1, follicles exhibited a concentration-dependent response (P < 0.05) to IGF-I, with the highest rates of granulosal cell proliferation, follicular integrity, and recovery rate of cumulus cell-oocyte complexes and lowest incidence of apoptosis occurring at the highest IGF-I dose. In Exp. 2 serum-free medium, granulosal cell proliferation was low (1 to 5%), irrespective of whether EGF and/or IGF-I were present and cellular apoptosis was increased (P < 0.05) on d 4 and 8 in the EGF+IGF-I group compared with the addition of either factor alone. In Exp. 3, granulosal cell proliferation was high in all follicles cultured in serum-containing medium for the first 3 d, but fell sharply (P < 0.05) on d 4, except in media containing IGF-I. Collectively, EGF and IGF-I increased granulosal cell proliferation, decreased apoptosis, and promoted follicular antrum formation. These results may provide useful information for developing a preantral follicular culture system in which the oocytes are capable of fertilization and embryonic development.     

Phage types, ribotypes and tetracycline resistance genes of Salmonella enterica subsp. enterica serovar Typhimurium strains isolated from different origins in Italy

The tetracycline resistance (tet) gene patterns of 52 tetracycline resistant Salmonella enterica subsp. enterica (S.) serovar Typhimurium isolates collected from animals, food of animal origin, and humans in Italy, were investigated to evaluate whether the tet gene patterns could be used for strain differentiation in addition to phage typing and ribotyping. The detection of tet genes was performed by specific PCR assays. Ribotyping was performed automatically using PvuII as restriction enzyme. Ten different ribotyping patterns were detected. All isolates were positive for at least one of the tet genes studied and six different tet gene patterns were observed. Ribotyping and tet gene patterns showed discriminatory indices of 0.741 and 0.812, respectively. Multiple tet genes were commonly found among tetracycline resistant S. typhimurium isolates from various sources. The resulting tet gene patterns allowed further discrimination of strains which were otherwise indistinguishable by their phage type, ribotype and origin. Thus, the analysis of tet gene patterns might represent an additional tool for the differentiation of S. typhimurium isolates.     

Immunohistochemical Detection of CD4-, CD8- and MHC II-Expressing Immune Cells and Endoglin in the Canine Corpus Luteum at Different Stages of Dioestrus

Specific immunohistochemical methods were applied to detect the presence of CD4-, CD8- and major histocompatibility complex II (MHC II)-expressing immune cells and of endoglin in the canine corpus luteum between days 15 and 75, after ovulation. Corpora lutea were obtained from groups of three clinically healthy beagle bitches, ovariohysterectomized at the respective days. For all four parameters, the effect of time was highly significant. Quantitative evaluation yielded high values on day 15, followed by a decrease on day 30 (CD4, CD8 and endoglin) and day 45 (MHC II). While there were no further changes for cells staining positive for CD4 and endoglin, CD8-positive immune cells increased from day 45 to day 60 to drop again on day 75; MHC II-positive staining increased from day 45 to days 60-75. These data suggest an involvement of the immune system in control of luteal function also in the dog that may have both stimulatory and inhibiting effects.     

Effects of supplemental vitamin E during the laying period on the reproductive performance of Taiwan native chickens

1. One-day-old Taiwan Native Breeder female chicks were fed on maize/soybean growing diets without supplemental vitamin E from hatch to 17 weeks of age. After 17 weeks the birds (n = 300) were randomly assigned to 5 dietary treatments and fed on maize/soybean laying diets supplemented with 0, 40, 80, 120 and 160 mg/kg of vitamin E (dl-alpha-tocopherol acetate), respectively, until 46 weeks of age. The variates measured included: age at first egg, feed consumption (FC), feed efficiency (FE), egg production (EP), egg weight (EW), egg specific gravity (ESG), eggshell strength (ESS), fertility and hatchability. 2. The addition of 120mg/kg of vitamin E lowered the first EW (P<0.05); however, there was no significant difference in the age or body weight (BW) of pullets at first egg or mortality rate to 46 weeks of age among the treatments. FE and egg mass were improved (P<0.05) in pullets fed 80 mg/kg of supplemental vitamin E. A significant increase in EP was observed after peak EP in pullets given 80 mg/kg of supplemental vitamin E. However, this favourable effect decreased as supplemental vitamin E exceeded 80 mg/kg. 3. From 17 to 46 weeks of age, egg quality (ESG and ESS) decreased with age. However, there was no correlation between age and fertility or hatchability during the experimental period, suggesting that egg quality is more age-sensitive than reproductive performance for breeder pullets. 4. Compared with the control, fertility and hatchability of all eggs set for the treatment with 80 mg/kg supplemental vitamin E increased by 7.7 and 13.4%, respectively. There was no difference in the hatchability of fertile eggs. 5. These results suggest that using supplemental vitamin E during the laying period can improve the reproductive performance of breeder pullets. The addition of 80 mg/kg of vitamin E obtained the best performance in EP, egg mass, FE (feed/egg), hatchability and fertility.     

DNA methylation and targeted sequencing of methyltransferases family genes in canine acute myeloid leukaemia,modelling human myeloid leukaemia

Tumours shows aberrant DNA methylation patterns, being hypermethylated or hypomethylated compared with normal tissues. In human acute myeloid leukaemia (hAML) mutations in DNA methyltransferase (DNMT3A) are associated to a more aggressive tumour behaviour. As AML is lethal in dogs, we defined global DNA methylation content, and screened the C‐terminal domain of DNMT3 family of genes for sequence variants in 39 canine acute myeloid leukaemia (cAML) cases. A heterogeneous pattern of DNA methylation was found among cAML samples, with subsets of cases being hypermethylated or hypomethylated compared with healthy controls; four recurrent single nucleotide variations (SNVs) were found in DNMT3L gene. Although SNVs were not directly correlated to whole genome DNA methylation levels, all hypomethylated cAML cases were homozygous for the deleterious mutation at p.Arg222Trp. This study contributes to understand genetic modifications of cAML, leading up to studies that will elucidate the role of methylome alterations in the pathogenesis of AML in dogs.     

Paediatric HUS Cases Related to the Consumption of Raw Milk Sold by Vending Machines in Italy: Quantitative Risk Assessment Based on Escherichia coli O157 Official Controls over 7 years

A quantitative risk assessment (RA) was developed to estimate haemolytic‐uremic syndrome (HUS) cases in paediatric population associated with the consumption of raw milk sold in vending machines in Italy. The historical national evolution of raw milk consumption phenomenon since 2008, when consumer interest started to grow, and after 7 years of marketing adjustment, is outlined. Exposure assessment was based on the official Shiga toxin‐producing Escherichia coli O157:H7 (STEC) microbiological records of raw milk samples from vending machines monitored by the regional Veterinary Authorities from 2008 to 2014, microbial growth during storage, consumption frequency of raw milk, serving size, consumption preference and age of consumers. The differential risk considered milk handled under regulation conditions (4°C throughout all phases) and the worst time–temperature field handling conditions detected. In case of boiling milk before consumption, we assumed that the risk of HUS is fixed at zero. The model estimates clearly show that the public health significance of HUS cases due to raw milk STEC contamination depends on the current variability surrounding the risk profile of the food and the consumer behaviour has more impact than milk storage scenario. The estimated HUS cases predicted by our model are roughly in line with the effective STEC O157‐associated HUS cases notified in Italy only when the proportion of consumers not boiling milk before consumption is assumed to be 1%. Raw milk consumption remains a source of E. coli O157:H7 for humans, but its overall relevance is likely to have subsided and significant caution should be exerted for temporal, geographical and consumers behaviour analysis. Health education programmes and regulatory actions are required to educate people, primarily children, on other STEC sources.     

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.     

Repair and Function of Synovium After Arthroscopic Synovectomy of the Dorsal Compartment of the Equine Antebrachiocarpal Joint

The reparative ability of equine synovium was determined by gross, histological, and ultrastructural examination. The functional potential of the synovium was estimated by examination of synovial cell organelles with transmission electron microscopy. Results from rested and exercised horses were compared to determine the effect of exercise on synovial healing. The response of the synovectomized joint to exercise was evaluated with a standardized lameness examination and by gross, histological, and histochemical observations of the articular cartilage. A 7-mm diameter motorized synovial resector was used to perform a subtotal synovectomy in 1 antebrachiocarpal joint of each of 8 horses; the contralateral joint served as a control. After 2 months rest, four randomly selected horses were rigorously exercised for the remainder of the study; the other four horses continued paddock rest. Lameness examinations and synovial fluid analyses were conducted at 0, 2, 30, 60, and 120 days. Synovium and articular cartilage from all horses were examined at necropsy at 120 days. None of the horses were lame during the study, and a transient synovitis occurred in the synovectomized joints. The hyaluronan concentration of treated joints decreased at 2 days but returned to normal by 60 days. Synovial fluid composition, including hyaluronan concentration, was unchanged by exercise. Significant cartilage damage was not observed in any of the joints. At 120 days, the healing synovium was devoid of villi and its subintima was fibrotic, however transmission electron microscopy confirmed that an intimal layer was present within the repair tissue. The cells within the repair tissue appeared actively engaged in both synthesis and phagocytosis. Exercise did not modify any of these findings. The results of this study suggest that 120 days after subtotal synovectomy, the joint environment was maintained and the resected synovium had evidence of restoration and increased metabolic potential. Synovectomized joints withstood exercise but synovial repair was not accelerated by exercise.     

Heat stress: A major contributor to poor animal welfare associated with long-haul live export voyages

Recent investigations by the Australian Department of Agriculture, Fisheries and Forestry into high mortalities on live export voyages from Australia to the Middle East during the Northern hemisphere summer suggest that animal welfare may be compromised by heat stress. The live export industry has generated a computer model that aims to assess the risk of heat stress and to contain mortality levels on live export ships below certain arbitrary limits. Although the model must be complied with under Australian law, it is not currently available for independent scientific scrutiny, and there is concern that model and the mandated space allowances are inadequate. This review appraises the relevant literature on heat stress in sheep and cattle, including laboratory studies aimed at mimicking the ambient temperatures and humidity levels likely to be encountered on live export voyages. Animal welfare is likely to be very poor as a result of heat stress in some shipments.