Acute phase protein response in goats

Acute phase proteins (APPs) are important diagnostic indicators of inflammatory disturbances in animals. The objectives of the current study were to validate analytical methods for measuring haptoglobin (Hp), serum amyloid A (SAA), acid soluble glycoprotein (ASG), fibrinogen, and albumin concentrations in goats and to evaluate their response to an inflammatory stimulus in this species. Intra- and interassay coefficients of variation (CVs) were in the range 0.07-9.31% and 1.83-12.68%, respectively, for all APPs and showed good precision. All assays determined APPs in a linear manner at different sample dilutions with high correlation coefficients with the exception of fibrinogen, which was measured by the heat precipitation method. Subcutaneous injection of turpentine oil induced an increase in Hp, SAA, ASG, and fibrinogen serum concentrations and a decrease in albumin concentration.     

Evaluation of haematological,serum biochemical and histopathological parameters of growing rabbits fed Amaranthus dubius

Amaranthus dubius is an alternative source of fibre and protein for the feed formulation for growing rabbits, although its effects on their health status are yet unknown. Therefore, this study aimed to evaluate the effects of three dietary levels (0, 160 and 320 g/kg) of A. dubius on haematological, serum biochemical and histopathological parameters of fattening rabbits. One hundred and thirteen New Zealand White rabbits were fed ad libitum from weaning (35 days) to slaughter (87 days). The health status of the animals was monitored, and mortality was recorded. Before slaughtering, 30 rabbits were randomly selected for haematological and serum biochemical analysis. During slaughter, 18 rabbits were randomly selected for histopathological study in heart, liver, lung and small intestine. The health status of rabbits was good, with no differences among treatments in mortality. Diet and sex did not affect the values of most haematological and serum biochemical parameters. Fibrinogen, glucose and high‐density lipoprotein values increased significantly from 0 to 320 g/kg A. dubius diets. The white blood cell count and glutamic oxaloacetic transaminase values decreased significantly from 0 to 320 g/kg A. dubius diets. The organs showed histologically well‐structured tissues. However, in all experimental groups several samples with lesions were observed, always highly focalized in small tissue areas. The main lesions observed, probably related to conditions of thermal stress affecting the rabbits, were cholesterolosis (heart), steatosis (liver), inflammatory lymphoplasmacytic infiltrate (lung) and chronic inflammatory tissue damage (small intestine). In conclusion, the inclusion of up to 320 g/kg A. dubius in diet did not jeopardize the health status of growing rabbits. Therefore, A. dubius can become an alternative raw material for the formulation of feeds in tropical and subtropical countries where this plant is widespread.     

Gene expression differences in the methionine remethylation and transsulphuration pathways under methionine restriction and recovery with D,L‐methionine or D,L‐HMTBA in meat‐type chickens

This study examined the molecular mechanisms of methionine pathways in meat‐type chickens where birds were provided with a diet deficient in methionine from 3 to 5 weeks of age. The birds on the deficient diet were then provided with a diet supplemented with either D,L‐methionine or D,L‐HMTBA from 5 to 7 weeks. The diet of the control birds was supplemented with L‐methionine from hatch till 7 weeks of age. We studied the mRNA expression of methionine adenosyltransferase 1, alpha, methionine adenosyltransferase 1, beta, 5‐methyltetrahydrofolate‐homocysteine methyltransferase, 5‐methyltetrahydrofolate‐homocysteine methyltransferase reductase, betaine‐homocysteine S‐methyltransferase, glycine N‐methyltransferase, S‐adenosyl‐L‐homocysteine hydrolase and cystathionine beta synthase genes in the liver, duodenum, Pectoralis (P.) major and the gastrocnemius muscle at 5 and 7 weeks. Feeding a diet deficient in dietary methionine affected body composition. Birds that were fed a methionine‐deficient diet expressed genes that indicated that remethylation occurred via the one‐carbon pathway in the liver and duodenum; however, in the P. major and the gastrocnemius muscles, gene expression levels suggested that homocysteine received methyl from both folate and betaine for remethylation. Birds who were switched from a methionine deficiency diet to one supplemented with either D,L‐methionine or D,L‐HMTBA showed a downregulation of all the genes studied in the liver. However, depending on the tissue or methionine form, either folate or betaine was elicited for remethylation. Thus, mRNA expressions show that genes in the remethylation and transsulphuration pathways were regulated according to tissue need, and there were some differences in the methionine form.     

Serotypes, virulence genes, intimin types and PFGE profiles of Escherichia coli isolated from piglets with diarrhoea in Slovakia

Two hundred and fifty Escherichia coli isolates from diarrhoeic and healthy piglets were serotyped and tested for the presence of virulence genes for fimbriae, intimin, heat-labile (LT) and heat-stable (STa and STb) enterotoxins, Stx toxins, and enteroaggregative heat-stable 1 (EAST1) enterotoxin by polymerase chain reaction (PCR). Although 220 isolates from diarrhoeic piglets belonged to 43 O serogroups and 77 O:H serotypes, 60% were of one of the 10 serogroups O2, O8, O15, O54, O84, O101, O141, O147, O149 and O157, and 60% belonged to only 10 serotypes (O8:H-, O54:H-, O84:H7, O101:H-, O141:H-, O141:H4, O147:H-, O149:H10, O163:H-, and ONT:H-). PCR showed that 79% of 220 isolates carried genes for at least one of the virulence factors tested. The gene encoding for EAST1 was the most prevalent (65%) followed by those encoding for STb (49%), LT (42%), STa (13%), and Stx2e (4%). Eighty-three (38%) of the 220 E. coli isolates carried the gene for F4 (K88), whereas genes for F18, F5 (K99), F41, F6 (P987), F17, and intimin (eae) were detected in 9%, 3%, 3%, 3%, 1%, and 3%, respectively. Seropathotype O149:H10:F4:LT/STb/EAST1 (70 isolates) was the most common, representing 32% of isolates. Pulsed-field gel electrophoresis (PFGE) analysis with XbaI of 15 O149:H10 representative isolates from diarrhoeic piglets distinguished 14 types. The 15 isolates exhibited a wide variability of distinct restriction patterns though all belonged to the same serotype (O149:H10), and all but one showed identical virulence determinants (F4, LT, STb, and EAST1). Among 30 isolates from healthy piglets only two virulence genes were detected: EAST1 (26%) and eae (17%). In total, 12 isolates were positives for the eae gene: five isolates had intimin beta1, four possessed intimin theta and three showed intimin type xiB. This is believed to be the first study describing the presence of intimin type xiB in E. coli of porcine origin.     

Application of VIS/NIR spectroscopy for estimating chemical,physical and mechanical properties of cork stoppers

The aim of this study was to evaluate the potential of visible and near infrared spectroscopy (Vis/NIRS) in predicting the chemical, physical and mechanical behavior of single-piece natural corks stoppers used for sealing wine bottles. Two training sets of 90 and 150 cork stoppers were used to obtain four spectra per sample in different positions: two of the stopper bases (transversal section) and two of the stopper sides (tangential section and radial section). The samples were scanned in the range of 400–2,500 nm using a Foss-NIRSystems 6500 SY II spectrophotometer equipped with a remote reflectance fiber-optic probe. On each training set, two-thirds of the samples were used to develop modified partial least square (MPLS) calibration equations, and the remaining one-third of the sample for the external validation of these MPLS equations. The best equations were obtained for the transversal section, which is the recommended one when applying Vis/NIRS technology to cork. The best results for the chemical composition were obtained for waxes and total polyphenols, showing coefficient of determination of the cross validation (r _cv ² ) values of 0.64 and 0.56 and coefficient of determination of the external validation (r _EV ² ) values of 0.53 and 0.55, respectively. The best equation for the physical and mechanical parameters was obtained for moisture content (r _cv ²  = 0.86 and r _EV ²  = 0.85), with somewhat lower results for density, compression force and extraction force (r _cv ²  = 0.66, 0.72, 0.52 and r _EV ²  = 0.52, 0.49, 0.51, respectively). The SECV (standard error of cross validation) and SEP (standard error of external validation) were similar for all the physical and mechanical parameters, thus confirming the robustness of the equations. MPLS model for moisture content fulfills the requirements for screening (RPD >2.5), but MPLS models obtained for waxes, total polyphenols, density, compression force and extraction force are not good enough for routine analysis or quality control. The results obtained from the MPLS models based on Vis/NIRS technology would permit the continuous quality control of humidity in the production line as well as obtaining information about certain chemical components (extractives contents) and some physical and mechanical parameters (density, extraction force and compression force).     

Placental thrombosis in acute phase abortions during experimental Toxoplasma gondii infection in sheep

After oral administration of ewes during mid gestation with 2000 freshly prepared sporulated oocysts of T. gondii isolate M4, abortions occurred between days 7 and 11 in 91.6% of pregnant and infected ewes. Afterwards, a further infection was carried out at late gestation in another group of sheep with 500 sporulated oocysts. Abortions happened again between days 9 and 11 post infection (pi) in 58.3% of the infected ewes. Classically, abortions in natural and experimental ovine toxoplasmosis usually occur one month after infection. Few experimental studies have reported the so-called acute phase abortions as early as 7 to 14 days after oral inoculation of oocysts, and pyrexia was proposed to be responsible for abortion, although the underline mechanism was not elucidated. In the present study, all placentas analysed from ewes suffering acute phase abortions showed infarcts and thrombosis in the caruncullar villi of the placentomes and ischemic lesions (periventricular leukomalacia) in the brain of some foetuses. The parasite was identified by PCR in samples from some placentomes of only one sheep, and no antigen was detected by immunohistochemical labelling. These findings suggest that the vascular lesions found in the placenta, and the consequent hypoxic damage to the foetus, could be associated to the occurrence of acute phase abortions. Although the pathogenesis of these lesions remains to be determined, the infectious dose or virulence of the isolate may play a role in their development.     

Modeling spatial distribution of European badger in arid landscapes: an ecosystem functioning approach

Understanding the factors determining the spatial distribution of species is a major challenge in ecology and conservation. This study tests the use of ecosystem functioning variables, derived from satellite imagery data, to explore their potential use in modeling the distribution of the European badger in Mediterranean arid environments. We found that the performance of distribution models was enhanced by the inclusion of variables derived from the Enhanced Vegetation Index (EVI), such as mean EVI (a proxy for primary production), the coefficient of variation of mean EVI (an indicator of seasonality), and the standard deviation of mean EVI (representing spatial heterogeneity of primary production). We also found that distributions predicted by remote sensing data were consistent with the ecological preferences of badger in those environments, which may be explained by the link between EVI-derived variables and the spatial and temporal variability of food resource availability. In conclusion, we suggest the incorporation of variables associated with ecosystem function into species modeling exercises as a useful tool for improving decision-making related to wildlife conservation and management.     

Evaluation of different enzyme-linked immunosorbent assays for the diagnosis of brucellosis due to Brucella melitensis in sheep

Six serological assays for the diagnosis of ovine brucellosis, due to Brucella melitensis were evaluated. Reference serum samples from sheep of known B. melitensis infection status (n = 118) were assessed using the Rose Bengal test (RBT), complement fixation test (CFT) and four commercial enzyme-linked immunosorbent assays (ELISAs), including two indirect ELISAs (iELISAs), one competitive ELISA (cELISA) and one blocking ELISA (bELISA).The highest differential positive rates (DPR) were obtained with the cELISA and bELISA, while the lowest DPR was estimated using iELISAs. A latent class analysis was performed to estimate the accuracy of the CFT, RBT and bELISA using 1827 sera from sheep undergoing testing as part of a surveillance and control programme. Lower sensitivity and specificity were obtained for the three serological tests when the field samples were used. A higher DPR was achieved by the CFT, compared to bELISA and RBT. The results suggest that ELISAs, and particularly the bELISA, might be suitable for inclusion in the European Union legislation on intra-community trade for diagnosing B. melitensis infection in sheep, as it has a similar test performance compared to the RBT.     

Canopy transpiration of a Pinus canariensis forest at the tree line: implications for its distribution under predicted climate warming

Canopy transpiration (E _c) of a 50-year-old Pinus canariensis Chr. Sm. Ex DC. stand at tree line in Tenerife, Canary Islands, was estimated continuously throughout a year from March 1, 2008, to February 28, 2009, by means of xylem sap flow measurements. E _c varied markedly throughout the entire year generally following the seasonal trends in soil water availability and varied between 0.89 mm day^?1 under the conditions of non-limiting soil water availability and close to zero under soil drought. This is because canopy conductance declined significantly with increasing evaporative demand and thus significantly reduced tree water loss, and this decrease was more pronounced during the soil drought. Total annual E _c was 79.6 mm, which is significantly below the values estimated for other Mediterranean forest ecosystems and even 70 % lower than the value estimated for a P. canariensis forest at 1,650 m a.s.l. where the soil water content was higher than at the tree line site. Therefore, these results highlighted the importance of drought stress in tree line ecotone and should be taken more into account in semiarid tree lines.