Molecular typing of Mycoplasma agalactiae: tracing European-wide genetic diversity and an endemic clonal population

Mycoplasma agalactiae causes chronic infections in small ruminants and remains endemic in many regions of the world, despite intensive and costly eradication programs. In this study, the innate genomic plasticity of M. agalactiae was exploited to design and assess a combination of molecular epidemiological tools to trace the pathogen in different geographic locations and to understand its emergence or re-emergence after eradication campaigns. For this purpose, two collections of M. agalactiae isolates, representing European outbreaks or localized endemic disease in a single region of France, were subjected to RFLP (Restriction Fragment Length Polymorphism) analyses using two sets of DNA probes (distributed across the genome and specific for the vpma gene locus), and a previously described VNTR (Variable Number Tandem Repeats) analysis. A combination of four genome-specific DNA probes and two VNTRs gave the highest discriminative power. Molecular typing revealed that, while isolates from diverse geographical origins fell into clearly different groups, the endemic disease repeatedly observed in the Western Pyrenees region over the past 30 years has been caused by a unique subtype of M. agalactiae. This indicates that the re-emergence of the pathogen after seemingly successful eradication programs is not due to the importation of exotic strains, but to the persistence of local reservoirs of infection.     

Evaluation of a multi‐agent chemotherapy protocol combining dexamethasone,melphalan, actinomycin D,and cytarabine for the treatment of resistant canine non‐Hodgkin high‐grade lymphomas: a single centre's experience

The DMAC protocol (dexamethasone, melphalan, actinomycin‐D, cytarabine) has been evaluated in American studies for the treatment of relapsed canine lymphoma, comparing similarly to other rescue protocols. The aim of this study was to evaluate efficacy and toxicity of DMAC, in a larger UK cohort of resistant canine lymphomas. Medical records of dogs with resistant non‐Hodgkin high‐grade lymphomas that received DMAC as a rescue protocol were reviewed from 2007 to 2017. Response, time from initiation to discontinuation (TTD) and toxicity (Veterinary Cooperative Oncology Group criteria) were assessed. One hundred dogs were included; 86 received CEOP (modified CHOP including epirubicin) as first‐line treatment. Thirty‐five dogs (35%) responded: 21 complete responders (CRs) and 14 partial responders (PRs). Responders had significantly longer TTD (P < 0.001) compared with non‐responders: 62 days (range 28‐952) for CR vs 32 days (range 20‐70) for PR. Six CR received more than six cycles of DMAC (range 7‐36 cycles) and experienced a longer TTD (median 508, range 126‐952 days). Thrombocytopenia occurred in 45% (24 grade 1‐2, 21 grade 3‐4) and neutropenia in 36% of cases (29 grade 1‐2, 7 grade 3‐4). Gastrointestinal toxicity occurred in 42% of dogs (40 grade 1‐2, 2 grade 3‐4). Owing to chemotherapy toxicity, treatment was discontinued in five, and hospitalization required in six cases. In this study, response to DMAC was lower and of generally shorter duration than previously reported. Toxicity was high, but infrequently led to hospitalization or discontinuation of treatment.     

Studies on the Nutrition of Spotted Sand Bass Paralabrax maculatofasciatus: Effect of the Dietary Protein Level on Growth and Protein Utilization in Juveniles Fed Semipurified Diets

Two feeding trials were conducted to determine the digestibility of a casein-based semi-purified diet and the effects of different protein levels on growth and protein use of spotted sand bass Paralabrax maculatofasciatus juveniles. For trial I, a semipurified diet with vitamin-free casein as the sole source of protein was fed three times a day to apparent satiation, for a period of 20 d. Feces were collected by siphoning each tank. The digestibility of the experimental diet was high: 97% for protein, 89% for lipids, and 84% for gross energy, whereas that of organic matter was 78%. For trial II, seven diets were formulated using vitamin-free casein at graded levels (25, 30, 35, 40, 45, 50, and 55% protein). Triplicate tanks for each dietary treatment were stocked with fish and fed by hand three times a day to apparent satiation for 6 wk. Perfomance of fish fed the different diets was evaluated for survival, percent weight gain, specific growth rate, feed conversion ratio, and protein efficiency ratio. Survival was 100% for all treatments. Growth of spotted sand bass juveniles increased as the dietary protein increased, but no evidence of reaching a plateau was found. The daily feed intake values showed an inverse relation to the protein content of the diets. The feed conversion ratio did not differ among diets containing 40% protein or greater. The results indicate that spotted sand bass juveniles with 2.5-g mean weight need at least 55% dietary protein for best growth when casein is the sole protein source. However, in terms of feed conversion ratio, the requirement apparently could be lower.     

A common near infrared—based partial least squares regression model for the prediction of wood density of Pinus pinaster and Larix?×?eurolepis

Wood density is defined as the ratio of mass to volume and therefore in principle it should be possible to calculate a unique partial least squares regression (PLS-R) model for several species. PLS-R models for wood density based on X-ray microdensity data were calculated for each species Pinus pinaster and Larix × eurolepis and for both species together. After cross-validation and test set validation the data sets were combined and final models were calculated. The common model gave a residual prediction deviation (RPD) of 3.1, a range error ratio (RER) of 11.7, and a SEP/SEC of 1.06. The single models for Pinus pinaster and Larix?×?eurolepis gave RPD’s of 3.5 and 3.2, RER’s of 13 and 11, and a SEP/SEC of 1.2. To the best knowledge of the authors all obtained PLS-R models are the first ones that fulfil the requirements according to AACC Method 39-00 (AACC in AACC Method, 39-00:15, 1999) to be used at least for screening (RPD?≥?2.5). Although this method and the defined limits were developed for the analysis of grains they can be used as a rough rule of thumb until limits for wood are available. The improvement of the PLS-R models, compared to published results, might be due to three facts (1) the higher number of scans collected for a single spectrum, (2) that the samples were better represented by the NIR spectra and X-ray microdensity values, and (3) that the sites for the measurement of NIR spectra and X-ray microdensity were coincided as strictly as possibly.     

Changes in green coffee protein profiles during roasting

To reveal its flavor, coffee has to be roasted. In fact, the green coffee bean contains all ingredients necessary for the later development of coffee flavor. It is now widely accepted that free amino acids and peptides are required for the generation of coffee aroma. However, the mechanisms leading to defined mixtures of free amino acids and peptides remain unknown. Information pertaining to the identification of precursor proteins is also lacking. To answer some of these questions, two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) was used to follow the fate of green coffee proteins. Two conditions were considered: roasting and incubation of green coffee suspensions at 37 degrees C. Coffee beans were observed to acquire the potential to spontaneously release H(2)O(2) upon polymerization of their proteins during roasting. Fragmentation of proteins was also observed. Conversely, H(2)O(2) was found to control polymerization and fragmentation of green coffee proteins in solution at 37 degrees C. Polymerization and fragmentation patterns under the two conditions were comparable. These observations suggest that the two conditions under study triggered, at least to some extent, similar biochemical mechanisms involving autoxidation. Throughout this study, a unique fragmentation cascade involving the 11S coffee storage protein was identified. Generated fragments shared an atypical staining behavior linked to their sensitivity to redox conditions.     

Epidemiological characteristics of rabies in South Korea from 1993 to 2001

In the nine years from 1993 to 2001, 210 cases of rabies were recorded in domestic animals in South Korea; 115 cattle, 94 dogs and one farmed deer were affected. The annual incidence of rabies cases increased to a peak of 64 in 1998, and then decreased to about 30 cases per year. The cases were confined to the northern part of Kyounggi and Kangwon provinces. One hundred and forty-six cases (69.5 per cent) occurred in Kyounggi and 64 cases (30.5 per cent) in Kangwon province, and about 82 per cent of them were confined to two counties in Kyounggi province (29 per cent in Paju and 28.1 per cent in Younchun) and to Chulwon county in Kangwon province (25.2 per cent). However, over several years the outbreaks gradually moved south and east in both Kyounggi and Kangwon provinces. There were more rabies cases in cattle than in dogs, suggesting that the disease is transmitted by the sylvatic cycle. To investigate the relationship between rabies in domestic animals and wild animals, 107 wild animals, including Korean raccoon dogs, badgers, weasels and feral cats, were tested for rabies; 21 of the 67 Korean raccoon dogs tested (31 per cent) were infected. The cases in domestic animals were most common in winter, from December to February, and least common in summer, from June to September.     

Cellular and humoral local immune responses in sheep experimentally infected with Oestrus ovis (Diptera: Oestridae)

Cellular and humoral local responses were investigated following repetitive artificial Oestrus ovis infections in lambs. The presence of larvae induced a huge local recruitment of either leucocytes (T and B lymphocytes, macrophages) or granulocytes (eosinophils, mast cells and globule leucocytes). This cellular response was more pronounced in the ethmoid and sinus (development sites of second and third instar larvae) than in the septum or turbinates where first instar larvae migrate. Infected lambs produced Oestrus ovis specific IgG and IgA antibodies in their mucus. This local humoral response was mainly directed against larval salivary gland antigens and not against larval digestive tract antigens. Compared to the control animals, the sinusal mucosa of infected animals was extremely thickened and the epithelium exhibited hyperplasia, metaplasia and eosinophilic exocytosis. The possible roles of these local immune responses in the regulation of O. ovis larvae populations in sheep are discussed.     

Partial sequencing and expression of genes involved in glucose metabolism in adipose tissues and skeletal muscle of healthy cats

Impaired insulin sensitivity is increasingly recognised in cats, but sequences of genes involved in insulin-signalling are largely undetermined in this species. In this study, extended feline mRNA sequences were determined for the adiponectin, glucose transporter-1 (GLUT1), GLUT4, peroxisome proliferative activated receptor-gamma1 (PPARgamma1), PPARgamma2, plasminogen activator inhibitor-1 (PAI-1), monocyte chemoattractant protein-1 (MCP-1) and insulin receptor genes. Conserved dog-specific primers identified from human-dog mRNA alignments were used to amplify feline cDNA in the polymerase chain reaction (PCR). The feline sequences determined by this method were used to design feline-specific primers suitable for real-time PCR for quantification of gene expression in insulin sensitive tissues of healthy cats. Partial sequences of feline mRNAs had 86-95% identity with dog and human genes. Expression of adiponectin, GLUT1, GLUT4, PPARgamma1, PPARgamma2, PAI-1 and insulin receptor mRNA was detected and quantified in subcutaneous and visceral fat and skeletal muscle, whereas MCP-1 mRNA was detected in adipose tissue but not in skeletal muscle. Further characterisation of genes related to glucose metabolism in cats will provide additional insights into insulin-signalling mechanisms in this species.     

A diet formulated from by‐products and indigenous foodstuffs for laying hens

1. The performance of six commercial stocks of laying hens fed on a diet composed of home‐produced foodstuffs, not usually used for human consumption, was compared with that of birds fed on a conventional diet.

2. The experimental diet was calculated to be low in energy and methionine and high in fibre by conventional standards. Birds ate up to 12% more of this diet, but the calculated intakes of energy and methionine were less than those of the corresponding controls.

3. White stocks fed on the experimental diet produced seven eggs less per bird than controls while the production of brown stocks was essentially unaffected.

4. The gross efficiency of energy utilisation for egg production was better with the experimental diet.     

Atmospheric methane and nitrous oxide of the Late Pleistocene from Antarctic ice cores

The European Project for Ice Coring in Antarctica Dome C ice core enables us to extend existing records of atmospheric methane (CH4) and nitrous oxide (N2O) back to 650,000 years before the present. A combined record of CH4 measured along the Dome C and the Vostok ice cores demonstrates, within the resolution of our measurements, that preindustrial concentrations over Antarctica have not exceeded 773 +/- 15 ppbv (parts per billion by volume) during the past 650,000 years. Before 420,000 years ago, when interglacials were cooler, maximum CH4 concentrations were only about 600 ppbv, similar to lower Holocene values. In contrast, the N2O record shows maximum concentrations of 278 +/- 7 ppbv, slightly higher than early Holocene values.