Use of Artemisia annua as a natural coccidiostat in free-range broilers and its effects on infection dynamics and performance

This work investigated the preventive effect of Artemisia annua L. dried leaves supplied as a botanical coccidiostat to two broiler genotypes reared in a Danish free-range system in a factorial experiment (two genotypes and ± supplement of dried A. annua leaves). The genotypes White Bresse L40, a pure slow-growing line, and Kosmos 8 Ross, a hybrid genotype with medium growing characteristics, were used. Broilers were raised indoor until 29-days-old and kept free of parasites. Twelve groups of 30 randomly selected broilers were placed in the range forming three replicates for each treatment combination. The paddocks were cultivated with a mix of grass and clover. A separate group of broilers was naturally infected with Eimeria spp. oocysts and five animals nominated as "seeders" were introduced to the above mentioned 12 groups, 10 days after its formation, with each group consisting of 35 animals per plot. This infection strategy was meant to imitate the transmission pathway observed at farm level. Ten individual birds from each of the 12 groups, in total 120 animals of mixed sex, were monitored twice weekly for 30 days for oocysts excretion. PCR of pooled faecal samples, oocyst morphology and localization upon necropsy were used to identify the Eimeria species involved in the infection. In general, broilers from both genotypes in the range coped well with a coccidia infection caused by Eimeria acervulina and Eimeria maxima as no clinical symptoms, or deaths, were reported during the experiment. In general, broilers supplemented with A. annua dried leaves showed a significantly (p<0.05) reduced number of excreted oocysts during the infection with no interaction to genotype. Females generally had a significantly higher shedding of oocysts than males (p<0.05). The overall body weight gain and the daily weight gain when infection was subdued showed a three-way interaction among genotype, sex and treatment - accounted mainly for the fact that Kosmos females responded positively to the Artemisia treatment while Kosmos males responded negatively, and only minor differences were found between sexes for the White Bresse genotype. In conclusion, supply of A. annua dried leaves as a botanical coccidiostat significantly reduced oocyst output in free ranged broilers and thus may form part of a strategy to prevent commercial losses.     

Liver disease in young rabbits infected by calicivirus through nasal and oral routes

Calicivirus infection causes rabbit haemorrhagic disease (RHD) that kills more than 90% of adult animals, whereas young rabbits are naturally resistant to this viral disease. It has been proposed that the different response of adult and young rabbits to calicivirus infection is due to absence of viral receptors in respiratory and digestive systems of young animals. We have searched for liver disease in 4-week-old rabbits inoculated with a calicivirus suspension by intranasal and oral routes. These young rabbits showed cell damage and mononuclear infiltration of the liver. The hepatic lesions were associated with mild to moderate increase in circulating transaminases. We conclude that the previously reported reduction of viral receptors in the epithelium of respiratory and digestive systems of young rabbits does not inhibit calicivirus from inducing liver disease in these hosts.     

Liver Enzymes and Ultrastructure in Rabbit Haemorrhagic Disease (RHD)

Rabbit haemorrhagic disease (RHD) is caused by a calicivirus infection that kills most adult rabbits 24–72 h after viral inoculation. Two liver enzymes (AST, aspartate aminotransferase, and ALT, alanine aminotransferase) were monitored in blood samples of calicivirus-infected rabbits during the short course of RHD. Values of AST were used to differentiate three stages of hepatocellular degeneration in RHD: mild (up to 20-fold increase in AST), moderate (150–200-fold elevation of AST) and severe (more than 1000-fold elevation in AST). Liver samples of rabbits from these three biochemical stages of hepatocellular degeneration of RHD were studied by transmission electron microscopy to define the fine structure of the hepatocytes. In the mild hepatocellular degeneration there was proliferation (microvesiculation) of the smooth endoplasmic reticulum and swelling of mitochondria into spheroid bodies with loss of cristae. In moderate hepatocellular degeneration, vacuolization of cytoplasm and mitochondrial damage continued to be present, and there was also formation of autophagic vesicles. In the severe hepatocellular degeneration of RHD, the altered mitochondria also showed loss of density of their matrix; rupture of cytoplasmic vacuoles led to the formation of large vesicles. Marked depletion of liver glycogen was also found in this late stage of RHD. These data offer a correlation between biochemical and cytological features of the liver during the hepatocellular degeneration of RHD.     

Development of a multiplex PCR assay for polymorphism analysis of Brucella suis biovars causing brucellosis in swine

Swine brucellosis is caused by the biovars 1, 2 and 3 of Brucella suis the identification of which up to now relies on microbiological tests lacking adequate specificity together with time consuming and expensive molecular procedures. Based on sequence variation of the omp2b gene, we have developed a four primer set multiplex PCR assay that was tested for polymorphism analysis of B. suis biovars causing brucellosis in swine. The assay exploits the single nucleotide polymorphisms found in omp2b gene of B. suis reference biovars which are conserved in 43 B. suis field isolates from different geographic origins and hosts. Three specific amplification patterns (S1, S2 and S3) were obtained for reference strains of B. suis biovars 1, 2 and 3, respectively. However, some B. suis field isolates identified as biovars 2 or 3 according AMOS-PCR, PCR-RFLP of omp31 and omp2 genes and classical bacteriological methods, resulted also in S1 patterns, limiting the typing usefulness of the method.     

Effects of experimental inoculation of bovine respiratory syncytial virus in different inbred mice lineages: establishment of a murine model for BRSV infection

Bovine respiratory syncytial virus (BRSV), a member of the subfamily Pneumovirinae, family Paramyxoviridae, is a major cause of respiratory disorders in young cattle. A number of studies were conducted to validate a reliable animal model for the infection, since BRSV inoculation on the natural host is costly and often unsuccessful. Unfortunately, after inoculation of BRSV in Balb/C mice, viral replication may be detected; however, evident pathological alterations are absent on the experimentally infected animals. In order to establish a mice model that could be used further for preliminary studies of pathological and immunological aspects of BRSV infection, three mice inbred lineages (Balb/C, A/J and C57BL6), possessing different genetic backgrounds, were tested about its susceptibility to the inoculation with BRSV. Animals were inoculated through the nasal and ocular routes and were observed after inoculation. At 7 days post-inoculation (dpi) animals were necropsied and virological (virus isolation and viral nucleic acid amplification) as well as histopathological examinations were performed. A/J and C57BL6 showed interstitial pneumonia, when compared to the Balb/C group. These findings shows that mice may constitute a suitable model for the study of BRSV infections, depending on the mice strain used for experimental inoculations.     

Digestive enzyme activity and nutrient digestibility in meagre (Argyrosomus regius) fed increasing levels of black soldier fly meal (Hermetia illucens)

The effect of partially replacing fishmeal (FM) by black soldier fly larvae meal (Hermetia meal—HM) in meagre (Argyrosomus regius) diet was evaluated for nutrient digestibility and digestive enzyme activity. For that purpose, triplicate groups of fish (18.0 ± 0.02 g) were fed during 48 days either a control diet (CTR), without HM, or one of three diets including 100, 200 and 300 g/kg of HM, replacing 17, 35 and 52% of FM, respectively. Apparent digestibility coefficients (ADCs) of dry matter, energy, protein, lysine, isoleucine, leucine, phenylalanine, alanine, glutamate, glycine and serine presented a decreasing response with increased HM inclusion. Chitin ADC was null, independent of dietary HM inclusion. Total alkaline protease activity increased while trypsin activity decreased with dietary HM inclusion. No intestinal chitinolytic activity was detected. Intestinal alkaline protease zymogram revealed nine bands with proteolytic activity against casein, with molecular weights ranging between 15 and 75 kDa. Anti‐protease activity in the intestine was not affected by dietary inclusion of HM compared to the CTR diet. Overall, it is concluded that replacement of up to 17% FM with HM (100 g/kg HM inclusion level) in meagre diets has no major adverse effects in diet digestibility and digestive enzyme activity.     

Assessment of soil contamination by potentially toxic elements in the aljustrel mining area in order to implement soil reclamation strategies

Land pollution due to past mining activities is a major environmental issue in many European countries. The Aljustrel mine (SW Portugal), located in the western sector of the Iberian Pyrite Belt (IBP) presents a negative visual and environmental impact as a consequence of the mining activity that has developed since the Roman era. Its impacts are also a restraint on the life quality of the population. The exposure of pyrite and other sulphides to air are responsible for the pollution observed in soils, surface water and stream sediments. This paper investigates the pollution load of potential toxic elements in soil samples collected around the Aljustrel mining area. The aim is to assess the levels of soil contamination with respect to average concentrations of toxic elements in the region and to understand the partitioning and availability of pollutants in the area. The results showed severe soil contamination (mainly As, Cd, Cu, Pb and Zn). The concentrations of As (up to 3936 mg kg⁻¹) and certain heavy metals (up to 5414 mg kg⁻¹ Cu, 61·6 mg kg⁻¹ Cd, 20 000 mg kg⁻¹ Pb and 20 000 mg kg⁻¹ Zn) are two orders of magnitude above the regional South Portuguese Zone (SPZ) background values. The median concentrations of As, Cd, Cu, Pb and Zn exceed the values established for world soils, the European Union, Portugal and Andalusia. The results suggest that the distribution patterns of Co, Cr and Ni element concentrations in the Aljustrel area are primarily influenced by the lithology and geochemistry nature of bedrock. The soil background of this geological domain is characterized by relatively high heavy metal contents, essentially derived from the parent rocks. Copyright © 2010 John Wiley & Sons, Ltd.     

Response to long‐term selection for egg production in laying hens

1. A White Leghorn line was selected for part‐record hen‐housed number of eggs from 1962 to 1990. Genetic changes were estimated as deviations from its unselected control line.

2. Over the first 10 generations with selection almost exclusively for number of eggs to the age of 273 d, all traits, except rate of mortality, showed significant changes. Regressions per year were: 273 d production, 3.07 eggs; 497 d production, 5.18 eggs; production from 274 to 497 d, 2.43 eggs; age at first egg, ‐2.33 d; mean weight of first 10 eggs, ‐0.82 g; body weight at 497 d, ‐19.02 g and rate of mortality, 0.19%.

3. Over the rest of the period increasing selection pressure for egg weight has been applied. This resulted in positive changes for this trait and no or small negative changes in egg number.

4. In general, heritabilities and genetic correlations did not change over the period of selection. The heritability of the main trait of selection, production to 273 d was 0.19 ± 0.04 and heritabilities of egg size traits about 0.50.

5. The genetic correlation between egg production to 273 d and mean weight of first 10 eggs was estimated as ‐0.37 ± 0.06 but from the observed response a realised genetic correlation of ‐0.97 was calculated.     

Sorption and leaching potential of herbicides on Brazilian soils

Sorption of the herbicides alachlor, atrazine, dicamba, hexazinone, imazethapyr, metsulfuron-methyl, nicosulfuron, simazine and sulfometuron-methyl was characterized on six Brazilian soils, using the batch equilibration method. In general, weak acid herbicides (dicamba, imazethapyr, metsulfuron-methyl, nicosulfuron and sulfometuron-methyl) were the least sorbed, whereas weak bases such as triazines and nonionic herbicides (alachlor) were the most sorbed. The Kd values found showed a significant correlation with soil organic carbon content (OC) for all herbicides except imazethapyr and nicosulfuron. Koc values showed a smaller variation among soils than Kd . To estimate the leaching potential, Koc and the ground-water ubiquity score (GUS) were used to calculate half-lives ( t_1/2 ) that would rank these herbicides as leachers or non-leachers. Comparison of calculated values to published values for t_1/2 demonstrated that sulfonylureas and hexazinone are leachers in all soils, alachlor is transitional, and atrazine, simazine and dicamba are leachers or transitional, depending on soil type. Results discussed in this paper provide background to prioritize herbicides or chemical groups that should be evaluated in field conditions with regard to their leaching potential to ground-water in tropical soils.     

Study of the disappearance of endosulfan,parathion, trichlorfon and pirimicarb from broccoli and portuguese cabbage

The influence of surface of exposure of brassicas to insecticide sprays has been assessed in two experiments conducted in small plots where endosulfan, parathion, trichlorfon and pirimicarb were applied to a flowering brassica, broccoli (Brassica oleracea L. var italica Plenck) and to a leafy brassica (Brassica oleracea L. var. tronchuda Bailey). The differences of surface exposure resulted in much higher deposits and subsequent residue levels in broccoli. The results stress the need to evaluate residue data on groups of brassicas of different surfaces of exposure to pesticide spray when granting registration petitions for brassicas in general in order to establish pre-harvest intervals.