GAL‐ and VIP‐Immunoreactive Nerve Structures in the Ileum and Large Intestine of Pigs with Dysentery

The morphology, neurochemistry and function of intramural nerve structures in the mammalian gastrointestinal tract are relatively well known, but in normal, healthy individuals. The present study was aimed at investigating the chemical coding of nerve structures in the wall of the ileum and large intestine in normal pigs (n = 3) and in pigs undergoing dysentery (n = 6). Dysentery was evoked by artificial infection of the clinically healthy animals per os with Brachyspira hyodysenteriae. All the animals were deeply anaesthetized and transcardially perfused with 4% paraformaldehyde. The cryostat sections of the intestines were processed for double‐labelling immunohistochemistry using antisera against PGP 9.5, GAL and VIP. In the intramural plexuses of the control pigs, the percentage of GAL‐immunoreactive (GAL‐IR) perykarya varied from 11% (descending colon) to 19% (centrifugal turns of the ascending colon) whereas in the dysenteric pigs, it was distinctly higher, reaching from 28% (ileum) up to 48% (cecum). In the control animals, the percentage of VIP‐IR neuronal somata varied from 3% (descending colon) to 19% (ileum). In dysenteric pigs, it was from 6% (descending colon) up to 28% (cecum). In the muscular coat (MC) and mucous membrane (MM) of the normal intestine, very numerous GAL‐ and VIP‐IR nerve fibres were observed. The nerve fibres in the myenteric plexus (MP) were even more numerous than those in the muscular coat while in the outer (OSP) and inner (ISP) submucous plexuses, they were less abundant. In the dysenteric pigs, the nerve fibres found in MC, MP and OSP were less numerous, whereas those observed in ISP and MM were more abundant than those in the control animals. The present results suggest that GAL and VIP are involved in the regulation of inflammatory processes developing in the porcine gastrointestinal tract during dysentery.     

Ovum Pick-up in Cycling and Lactating Postpartum Swamp Buffaloes (Bubalis bubalis)

The objective of this study was to evaluate the efficiency of Ovum Pick Up (OPU) in cycling (n = 5) and lactating, postpartum, swamp buffaloes (n = 6) with and without gonadotropin stimulation. The OPU was performed every two weeks in all groups of animals, for a total of six sessions. Thirty collections were performed in five cycling buffaloes and 36 collections in six lactating postpartum buffaloes. Buffaloes that received hormonal stimulation were given a total of 400 mg, follicle stimulating hormone (FSH), administered twice daily over 3 days in decreasing doses, together with 100 microg of GnRH, 24 h after the last FSH injection. Following a resting period of 1 month, the two groups of buffaloes, were subjected to the same OPU regimen, but without any hormonal treatment for an additional six OPU sessions. The number of aspirated follicles recorded from the hormonal stimulated, cycling animals and lactating, postpartum buffaloes was not significantly different, 7.2 +/- 3.7 and 9.0 +/- 3.2, respectively (p > 0.05). Recovered oocytes collected from the two groups of hormonally stimulated animals were also not statistically different: 3.7 +/- 2.7 in the cycling and 5.9 +/- 3.5 in the lactating postpartum group (p > 0.05). In the two groups of buffaloes not receiving hormonal stimulation, the number of aspirated follicles was not significantly different: 2.1 +/- 1.4 and 1.4 +/- 0.7 in cycling and lactating postpartum buffaloes respectively (p > 0.05). Recovered oocytes in the non-treated groups were also similar: 1.4 +/- 1.3 vs 0.7 +/- 0.8 in cycling and lactating buffaloes (p > 0.05). Among stimulated buffaloes, most aspirated follicles were small in size (< or =5 mm), whereas they were mostly medium and large sizes in the non-treated buffaloes. The oocyte recovery rate in both the groups, cycling and lactating postpartum, were 51.6% and 69.5% in stimulated groups and 55.0% and 53.1% in non-stimulated groups (p > 0.05). The majority of recovered oocytes were single- and multi-layered, and the number was greater in the cycling than in the lactating, postpartum buffaloes. The number and quality of recovered oocytes was similar in all groups of buffaloes whether they were received or did not receive hormonal stimulation. Moreover no difference was found in multi- and single-layered oocytes between cycling and lactating, postpartum buffaloes. In conclusion, OPU can be performed successfully in swamp buffalo in different reproductive status and FSH administration was shown to increase the number of aspirated oocytes in both cycling and lactating, postpartum buffaloes.     

Anti‐microbial Susceptibility of Streptococcus spp. Isolated from Bovine Mastitis in Argentina

The in vitro susceptibility to penicillin G, erythromycin and clindamycin was determined by the disc diffusion test and by E‐test for a total of 47 streptococcal strains (three Streptococcus uberis, 36 Streptococcus agalactiae, eight Streptococcus dysgalactiae spp. dysgalactiae) isolated from bovine intramammary infections in Argentina. Moreover, resistance phenotypes of erythromycin‐resistant streptococcal isolates was characterized. MIC₉₀ of penicillin G, erythromycin and clindamycin for S. agalactiae were 0.75, 8.0 and 12.0 μg/ml respectively. Resistance to erythromycin and clindamycin was detected in 13 (27.6%) and 12 (25.5%) isolates respectively. No isolate was resistant to penicillin G. Resistance against macrolides, lincosamides and streptogramin B (MLS_B) represented by the constitutive MLS_B phenotype was present in 11 (23.4%) erythromycin‐resistant isolates and two isolates (4.3%) expressed the M phenotype. The inducible MLS_B phenotype was not identified. Results suggest that beta‐lactams are the first‐line antibiotics when treating streptococcal udder infections; however, the continuous monitoring of the antibiotic resistance is essential, as the emergence of resistant strains has become a growing concern on the therapy of bovine mastitis.     

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.     

Occurrence and Distribution of Lactate Dehydrogenase in Knee Joint Carilage in Healthy and Osteoarthrotic Dogs

According to clinical studies, degenerative diseases of canine joints lead to higher lactate dehydrogenase (LDH) levels in synovial fluid. The goal of the present study was to examine the intraarticular distribution of LDH in healthy and osteoarthrotic knee joints in order to identify possible sources of LDH in synovial fluid. As synovial LDH concentrations neither correlate with the number of leukocytes nor with synovitis, our investigation focused on the articular cartilage. Samples from healthy and osteoarthrotic knee joints were fixed and processed for transmission electron microscopy (TEM), immunohistochemistry (IHC), and immunocytochemistry (ICC). In addition, fresh cartilage samples were investigated cytochemically by the tetrazolium‐formazan reaction. Analyses of blood and synovial fluid samples were used to confirm the absence of inflammatory disease. Morphology of articular cartilage was assessed macroscopically and by means of TEM. IHC revealed highest levels of LDH in chondrones and a diffuse labelling of the matrix with a distinctive decrease in signal from superficial to deeper cartilage layers. Ultrastructural localization by ICC showed LDH to be present in the cytoplasm of all chondrocytes and confirmed the density gradient in the matrix. Labelling was absent from nuclei and from pericellular rims. Cytochemistry confirmed the distribution pattern and, thus, expanded our findings beyond immunological evidence by providing proof of enzymatic activity of LDH in articular cartilage. The present results indicate that LDH is transferred from chondrocytes to the cartilaginous matrix. We suggest, therefore, that LDH found in synovial fluid originates from the articular cartilage and that osteoarthrotic processes promote LDH release from the cartilaginous matrix.     

Assessing the potential impacts of alternative landscape designs on amphibian population dynamics

An individual-based, spatially explicit population model was used to predict the consequences of future land-use alternatives for populations of four amphibian species in two central Iowa (midwest USA) agricultural watersheds. The model included both breeding and upland habitat and incorporated effects of climatic variation and demographic stochasticity. Data requirements of the model include life history characteristics, dispersal behavior, habitat affinities, as well as land use and landcover in geographic information systems databases. Future scenarios were ranked according to change in breeder abundance, saturation, and distribution, compared to baseline conditions. Sensitivity of simulation results to changes in model parameters was also examined. Simulated results suggest that while all four species modeled are likely to persist under present and future scenario conditions, two may be more at risk from future landscape change. Although the study species are all widespread generalists regarded as having a low conservation priority, they depend on wetlands and ponds, increasingly endangered habitats in agricultural landscapes. Broader conservation strategies in the region would ensure that these currently common organisms do not become the endangered species of the future.This revised version was published online in May 2005 with corrections to the Cover Date.     

Development of Stem-base Pathogens on Different Cultivars of Winter Wheat Determined by Quantitative PCR

The progress of development of stem-base pathogens in crops of second winter wheat was plotted in nine experiments in three years. The amount of each pathogen present was determined by quantitative PCR. Where Tapesia yallundae was present in quantifiable amounts, it usually developed earlier than the other eyespot pathogen, T. acuformis. Both species were usually present in greater amounts on cultivars which are more susceptible to eyespot. The sharp eyespot pathogen, Rhizoctonia cerealis, developed more erratically than either of the Tapesia spp. and there were no consistent effects on different cultivars. Fusarium spp., the cause of brown foot rot, were rarely present in quantifiable amounts, but Microdochium nivale was usually present as one or both of the varieties nivale and majus. Late-season (after anthesis) decreases in M. nivale suggest that any brown foot rot symptoms attributable to this fungus would have fully developed earlier. Cultivar differences in amounts of M. nivale were most clear in stems during internode extension and when relatively large amounts of DNA were present. Such differences approximately reflected eyespot susceptibility, cv. Soissons containing most and cv. Lynx containing least DNA. The results emphasise the difficulty in relating diagnoses, by quantitative PCR or other means, at early growth stages when decisions to apply fungicides against stem-base disease are made, to later disease severity.     

Biological and Physical Constraints on Maize Production in the Humid Forest and Western Highlands of Cameroon

The aim was to identify biological and physical factors responsible for reducing maize yield in Cameroon. Two surveys were conducted in 137 fields in two agroecological zones in 1995–1997. In the Humid Forest (HF), Bipolaris maydis, Stenocarpella macrospora, Puccinia polysora, Rhizoctonia solani and soil fertility were factors that reduced maize production in 1995 and 1996. In the Western Highlands (WHL), Cercospora zeae-maydis, and the interaction between soil fertility and maize variety were the most important constraints to maize production in 1996. In 1997, C. zeae-maydis, S. macrospora, physiological spot and stem borer damage (Busseola fusca) were negatively related to ear weight. The combination of these biological factors (diseases and insects), and the physical parameter of soil fertility were responsible for reducing maize yield in these selected benchmarks of Cameroon. Maximum potential yield reductions were estimated at 68% due to B. maydis and 46% due to S. macrospora, respectively, in the HF in 1995. In 1996, maximum potential yield reductions in the HF were estimated at 34%, 41% and 30% due to S. macrospora, P. polysora and R. solani, respectively. In the WHL, C. zeae-maydis had the potential to cause a yield reduction of 79% in 1996. In the WHL in 1997, the interaction between C. zeae-maydis and B. fusca, stem diseases and the physiological spot caused potential reductions of 52%, 34% and 39%, respectively.