Changes in the plasma concentrations of prolactin,luteinising hormone,progesterone and d‐(β)‐hydroxybutyrate in turkey hens (meleagris gallopavo), during treatment of broodiness under commercial conditions

1. An experiment was done under commercial conditions to investigate the physiological effects of isolating broody turkey hens, for 72 h, in sand and wire floored pens on the third, 10th and 16th weeks of production.

2. Hens identified as broody and removed from the flock had higher plasma prolactin concentrations than the laying hens at each of the three experimental stages.

3. Confinement in sand and wire floored pens, induced a decline in plasma prolactin concentrations. This decline probably impeded immediate development of broody behaviour. Alternately, levels of prolactin higher than those of laying hens were again measured 7 and 14 days after treatment during third week but not after the 10 th and 16th week of production.

4. Confinement did not induce consistent changes in luteinising hormone (LH) and progesterone concentrations from one period to an other.

5. An increase in the plasma concentration of D‐(/?)‐hydroxybutyrate was observed in the hens which had an egg present in the oviduct on day 2, 3 and/or 4 of the treatment. Subsequently, a decrease in ovulation rate was observed in the hens with higher concentrations of D‐(β)‐hydroxybutyrate while under treatment, during the 10th week of production.

6. These data confirm that the effectiveness of the traditional methods for broodiness prevention under commercial conditions is related to the induction of a decrease of prolactin.     

Blood biological constants in the deer Rusa (Cervus timorensis russa) in New-Caledonia. I. Hematologic constants

Since the beginning of year 1987, the deer "Rusa" breeding has been developing in New Caledonia. In 1988, during a slaughter operation amidst the herds, nearly ninety blood samples were collected in order to define the blood biological parameters (or constants) of this species. Regarding haematology, the study concerns the following parameters: erythrocyte count (9.32 x 10(12)/l), leucocyte count (4.51 x 10(9)/l), various leucocyte lines and their formula, i.e. (neutrophile polymorphonuclear: 2.08 x 10(9)/l [46.6 p. 100]; lymphocytes: 1.75 x 10(9)/l [38.4 p. 100]; monocytes: 0.33 x 10(9)/l [7.5 p. 100]; eosinophiles polymorphonuclear leucocytes: 0.4 x 10(9)/l [7.46 p. 100]; basophile polymorphonuclear leucocytes: 0.01 x 10(9)/l [0.28 p. 100]), hematocrite (36.8 l/l), hemoglobin ratio (14.1 g/dl), mean corpuscular volume (40.3 dl), mean corpuscular hemoglobin rate (15.3 pg/cell), mean corpuscular hemoglobin concentration (38.7 g/dl). In the course of the study, variations of these parameters were detected according to various physiological criteria and to the sampling conditions as deer is a stress sensitive animal.     

Pregnancy Rates in Repeat-breeder Heifers Following Multiple Artificial Inseminations during Spontaneous Oestrus

Hormonal asynchronies during oestrus, related to the presence of suprabasal plasma-progesterone (P4) concentrations and a delayed ovulation, interfere with the fertility of repeat-breeder heifers (RBH). Since tubal dysfunction can occur in connection with hormonal asynchronies and constrained availability of fertile spermatozoa at the time of ovulation, the present study tested the hypothesis that frequent sperm deposition from onset of oestrus to ovulation may improve pregnancy rates in RBH. Five RBH and five virgin heifers (VH; controls) were repeatedly artificially inseminated (AI) at 6 h intervals from onset of oestrus to spontaneous ovulation. Hormone analyses revealed suprabasal P₄ concentrations and a delay in the occurrence of the luteinising hormone (LH) surge, but a normal cortisol profile in RBH. Compared with controls, RBH presented longer interval from onset of oestrus to ovulation, and therefore, received more AIs. Pregnancy rates in RBH reached control levels (60%; NS), indicating that the hypothesis might be correct. Pregnancy rates in VH were below the expected range, presumably attributed to a deleterious influence of the frequent handling. The study suggests that pregnancy rates can be improved in RBH by frequent AI in relation to spontaneous ovulation. However, this practice of repeated manipulations, while seeming not to show adverse effects, lacks practicality for routine use.     

Culture of Differentiated Adult Rabbit Auricular Chondrocytes

Chondrocytes dedifferentiate to a fibroblast‐like phenotype on plastic surfaces. Dedifferentiation is reversible if these cells are then cultured embedded in gels as alginate, agarose or collagen. Chondrocytes cultured in suspension on a non‐adherent surface are also known to form aggregates of differentiated cells. The knowledge of chondrocyte behavior in culture is relevant for tissue engineering purposes. In this report we describe a simple method to culture differentiated or redifferentiated rabbit auricular chondrocytes on plastic surfaces with a stable phenotype. When chondrocyte aggregates formed in suspension are next seeded on plastic surfaces, most of them attach to the plastic as round or polygonal cells, and this morphological differentiation, confirmed by the presence of type II collagen, is stable for long culture periods. We also report that the addition of aggregates to monolayer cultures of dedifferentiated chondrocytes results in their redifferentiation, as is shown by their morphological changes and the synthesis of type II collagen. Therefore, this simple method can be useful for the study of chondrocyte behavior on plastic surfaces and for redifferentiating previously proliferated chondrocytes in tissue engineering techniques. Furthermore, these results demonstrate that, in addition to culture conditions such as cell isolation method or cell‐density, chondrocyte behavior on plastic depends on the presence or absence of aggregates resulting from the dissociation process.     

The effects of agricultural fields and human settlements on the use of rivers by wildlife in the mid-Zambezi valley,Zimbabwe

After the eradication of the Tse-Tse fly in the Mid-Zambezi valley, human settlements and fields extended mainly along the main rivers. In order to investigate the consequences of this human development on wildlife diversity we monitored three rivers of the Mid-Zambezi valley in Zimbabwe: Angwa, Manyame and Kadzi. The rivers were divided in segments of 200 m which were checked for spoors in order to assess the number of species and the number of individuals that used the segments. Human settlements were also recorded. We used a GIS to define the spatial characteristics of the fields present along the rivers, and related them to the distribution and abundance of wild species spoors in the river beds and banks. Our results show that the number of species in one segment of the river decreased with the increasing size of the field area bordering the segment. For all the major ungulate species, the numbers of individuals recorded per segment decreased with increasing field area. A similar trend was observed for small and medium-sized carnivores, though they were in lower numbers when present. Our analyses thus confirm that the extension of human agriculture in wildlife areas has an impact on most wild species, but we also define some threshold value of field size above which there seem to be an acceleration of the decrease in wildlife density and diversity: 3.2 ha for medium and small herbivores and carnivores; only the elephant seem to tolerate larger field area with a threshold value of 32 ha.This revised version was published online in May 2005 with corrections to the Cover Date.     

Detection of the Defoliating Pathotype of Verticillium dahliae in Infected Olive Plants by Nested PCR

Spread of Verticillium wilt into newly established olive orchards in Andalucía, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants.     

Gene Flow Analysis of Phytophthora porri Reveals a New Species: Phytophthora brassicae Sp. Nov.

Isozyme analysis and sequence analysis of the internal transcribed spacer regions (ITS-1 and ITS-2) and the 5.8S subunit of the ribosomal DNA gene repeat were used to examine whether isolates of Phytophthora porri from Allium and Brassica represent a single homogeneous species. Twenty-six strains of P. porri, 16 strains isolated from the genus Allium, and 10 strains isolated from the genus Brassica, were analyzed using malate dehydrogenase (MDH), isocitrate dehydrogenase (IDH) and lactate dehydrogenase (LDH), represented altogether by four putative loci (Mdh-2, Idh-1, Idh-2, and Ldh-2). Isozyme analysis revealed that strains isolated from Allium contained five private alleles at three isozyme loci (Ldh-2 ⁸³, Ldh-2 ¹⁰⁴, Idh-1 ¹⁰⁸, Idh-1 ¹¹², and Idh-2 ⁹⁸), whereas six different alleles were observed at four isozyme loci (Ldh-2 ⁸⁵, Ldh-2 ¹⁰⁰, Ldh-2 ¹¹⁴, Idh-1 ¹⁰⁰, Idh-2 ¹⁰⁰, and Mdh-2 ¹¹¹) in strains obtained from Brassica. The heterozygosity at the Ldh-2 locus, differing in allele composition, however, between strains from Allium and Brassica, was present in all strains, indicating that it is probably fixed. Sequence analysis of the ITS regions and the 5.8S subunit showed consistent differences between isolates from Allium and isolates from Brassica. Based on isozyme data, ITS sequence analysis and formerly published differences in restriction enzyme patterns of mitochondrial DNA, morphology and pathogenicity, it was concluded that the isolates of P. porri Foister did not represent a homogeneous species. Isolates from Brassica constitute a distinct species which is described here as P. brassicae sp. nov. It was inferred from isozyme patterns, which were in no case intermediate between the two species, that P. porri and P. brassicae do not hybridize and are reproductively isolated by barriers to gene flow.     

Comparison of Crinipellis perniciosa isolates from Brazil by ERIC repetitive element sequence-based PCR genomic fingerprinting

Fifty isolates of Crinipellis perniciosa originating from Theobroma cacao , Heteropterys acutifolia and Solanum lycocarpum , from six states within Brazil, were characterized through ERIC-PCR, representing the first application of this method for molecular characterization within C. perniciosa . Phenetic analysis of banding patterns revealed a separation of isolates on the basis of host of origin, with T. cacao -derived isolates showing only a 0·2 similarity level to a cluster comprising the isolates from H. acutifolia and S. lycocarpum . Considerable intraspecific variability was observed within C. perniciosa isolates from T. cacao , with distinct groups observed correlating with geographical origin. Given that a number of isolates from T. cacao from the Amazon region grouped with isolates from Bahia state, this work discusses the possibility that current C. perniciosa populations pathogenic on T. cacao in Bahia originated from the Amazon region, rather than from alternative host plants.     

Identification ofTomato Mottle Taino Begomovirus strains in Cuban potato fields

The presence of a begomovirus in potato plants with yellow mottle symptoms was determined for the first time in Cuba. The incidence of typical begomovirus-like symptoms in potato plants in some regions of Havana province (Güira de Melena, San José de las Lajas, Güines and Boyeros) during the growing seasons from 1992 to 1998 was in general low. However, in some cultivars belonging to the National Program for Potato Genetic Improvement, the incidence reached 100%. Yield losses, determined in 1992 and 1994, ranged as high as 19% to 56.33% depending on the cultivar. Characterization of the causal agent was done by light microscopy, host range (graft and mechanical transmission), DNA hybridizations, polymerase chain reaction, and restriction fragment length polymorphism analysis. Nucleotide sequence of the amplified fragments revealed the presence ofTomato mottle Taino virus. The virus was transmittedvia tubers and has been detected in mixed infections withPotato virus X and withPotato leaf roll virus. http://www.phytoparasitica.org posting Oct. 20, 2003. The first two authors contributed equally to this work.     

Evidence that a Leaf-Disk Test Allows Assessment of Isolate-Specific Resistance in Brassica oleracea Crops Against Downy Mildew (Peronospora parasitica)

A rapid resistance/susceptibility test for Peronospora parasitica (downy mildew) was established by inoculating leaf-disks of four Brassica oleracea accessions. Several conditions were tested: disk disinfection or not, agar medium with or without nutrients and with 50 or 100 ppm of benzimidazole. Using disinfected disks placed on agar (no nutrient and benzimidazole at 50 or 100 ppm), the responses of leaf-disks to four isolates were similar to those obtained using the classical cotyledon test, whereas undesired contaminations occurred in all other conditions. The possible effect of the particular leaf used for obtaining the disks was also studied. In each incompatible interaction tested, disks were resistant whatever the leaf used. In compatible interactions, susceptible phenotypes were observed on disks derived from the six lowest leaves, but disks from upper leaves were resistant. The genetic basis of resistance in a F1 hybrid broccoli was assessed, by testing six isolates on an F2 population derived from this hybrid. The cotyledon test only allows inoculation of two isolates per seedling, whereas many isolates can be tested on each plant by using leaf-disks. The segregation of the resistance to each of the six isolates was analysed: two dominant genes (tightly linked) control resistance to all isolates (one to five isolates; the other to only one isolate).