Diversity and abundance of arbuscular mycorrhizal fungi spores under different coffee production systems and in a tropical montane cloud forest patch in Veracruz, Mexico

We evaluate the arbuscular mycorrhizal fungi (AMF) community as measured by spores in different coffee production systems (at the depth of 0?C15?cm). In addition, we analyze the similarities between the AMF communities in coffee production systems and those that occur in a tropical montane cloud forest patch in order to evaluate the capacity of coffee production systems to preserve the native AMF community. We carried out four samplings in five coffee production systems representative of a vegetation structure gradient, and in a forest. From 120 soil samples, 33 morphospecies were detected. In all the sites, the dominant morphospecies were Glomus clarum and Glomus sp. 3. We found no significant difference in AMF spore richness between sites. Diversity was similar in most of the coffee production systems. Significant differences were only detected in spore abundance; during the dry season the forest, shaded traditional rustic system and shaded simple system presented the highest spore abundance. With the exception of one species exclusive to the forest, the coffee production systems all share the same AMF species as the forest. The coffee production systems with the greatest similarity to cloud forest were the shaded traditional rustic system and the shaded simple system. It is suggested that control of weeds and fertilization could be important factors influencing the composition and abundance of AMF spores in coffee production systems.     

Exposure to Sarcocystis spp. in horses from Spain determined by Western blot analysis using Sarcocystis neurona merozoites as heterologous antigen

Horses serve as an intermediate host for several species of Sarcocystis, all of which utilize canids as the definitive host. Sarcocystis spp. infection and formation of latent sarcocysts in horses often appears to be subclinical, but morbidity can occur, especially when the parasite burden is large. A serological survey was conducted to determine the presence of antibodies against Sarcocystis spp. in seemingly healthy horses from the Galicia region of Spain. Western blot analyses using Sarcocystis neurona merozoites as heterologous antigen suggested greater than 80% seroprevalance of Sarcocystis spp. in a sample set of 138 horses. The serum samples were further tested with enzyme-linked immunosorbent assays (ELISAs) based on recombinant S. neurona-specific surface antigens (rSnSAGs). As expected for horses from the Eastern Hemisphere, less than 4% of the serum samples were positive when analyzed with either the rSnSAG2 or the rSnSAG4/3 ELISAs. An additional 246 horses were tested using the rSnSAG2 ELISA, which revealed that less than 3% of the 384 samples were seropositive. Collectively, the results of this serologic study suggested that a large proportion of horses from this region of Spain are exposed to Sarcocystis spp. Furthermore, the anti-Sarcocystis seroreactivity in these European horses could be clearly distinguished from anti-S. neurona antibodies using the rSnSAG2 and rSnSAG4/3 ELISAs.     

Phylogeographic patterns and possible incipient domestication of <Emphasis Type="Italic">Jacaratia mexicana</Emphasis> A. DC. (Caricaceae) in Mexico

Jacaratia mexicana A. DC. (Caricaceae) is a tropical tree distributed throughout Mexico and Mesoamerica. Some evidence in Mexico indicates the presence of an incipient domestication process in this species. Phylogeographical analyses can potentially determine contemporary patterns of gene flow, isolation between population lineages, as well as historical processes such as population bottlenecks or expansions on their geographical areas. In this study we reconstruct the phylogeographical patterns in populations of J. mexicana A. DC., in order to find differences between genetic variation among wild and cultivated populations utilizing chloroplast DNA and nuclear DNA sequences. We generate a Bayesian phylogenetic tree, to estimate the divergence time between clades using calibrated mutation rates. We also infer the demographic history of these populations using neutrality tests among wild and cultivated accessions. We identified higher levels of haplotype and nucleotide diversity for the cpDNA and ITS types in wild populations than in domesticated populations. These results indicate a reduction of genetic diversity derived from human selection on domestication traits. Neutrality test suggests population expansion detected by the significant negative values of Fu’s Fs in the cultivated populations of this specie. These process results in an excess of rare polymorphism with the fixation of certain advantageous mutation throughout time, this implication are in accordance with the role of the strong selection in the fruit traits of J. mexicana. The dated phylogeny constructed with BEAST program indicated a dispersion pattern for the J. mexicana ancestors across the South Pacific and South Eastern populations during the late Pliocene. Posterior dispersion and divergence in the clades from Central Mexico and North Pacific are in agreement with the episodes of mountain-building in different regions of Mexico.     

Molecular differentiation between NS1 gene of a field strain Bluetongue virus serotype 2 (BTV-2) and NS1 gene of an attenuated BTV-2 vaccine

At the end of September 2000, clinical symptoms of Bluetongue appeared in sheep flocks of the Balearic Islands (Spain). The presence of the BTV serotype 2 in tissue and blood samples of affected animals was confirmed by laboratory techniques. A systematic vaccination were carried out in affected areas using a live monovalent serotype 2 vaccine available from Onderstepoort laboratory (South Africa). In order to perform epidemiological studies, a new method to differentiate between the NS1 genes of BTV-2 affecting the Balearic islands and that of the Onderstepoort commercial live virus vaccine (monovalent, serotype 2) has been developed. This procedure is based on the use of an RT-PCR, followed by restriction endonuclease analysis. Epidemiological data of a study carried out in the period January-October 2001 using this procedure are included.     

Fungi isolated from bovine udders,and their possible sources

AIM: To identify fungi isolated from infections of the bovine mammary gland, and establish their possible sources.

METHODS: From a herd of 420 cows, milk samples were collected from all quarters at calving and cultured to detect causative organisms. Quarters identified as infected with fungi were further sampled during early lactation. Samples from feedstuffs, the feed pad and ends of teats were also collected and analysed for the presence of fungi.

RESULTS: Eleven of 420 cows were diagnosed with intramammary infections (IMI) caused by yeasts (nine cows, 10 quarters) and moulds (two cows, three quarters). Six of the yeast species had previously been reported as being responsible for mastitis. Elevated somatic cell counts (SCC) were observed in many quarters, but most infections were eliminated spontaneously. Two of the fungi isolated from milk samples were also isolated from feedstuffs and teat swabs, and seven other fungi isolated from milk samples were not isolated from feed, the feed pad or cows' teats.

CONCLUSIONS: Isolation of fungi from the udder is rarely reported in dairy cows in New Zealand. In this herd, contamination of the end of the teat originating from feedstuffs and possibly exacerbated by the use of a feed pad may have led to the establishment of IMI caused by fungi.

CLINICAL RELEVENCE: Fungi are infrequently if ever reported in mastitis trial data or surveys in New Zealand and are probably of little clinical significance.     

Assessment of Different Functional Parameters of Frozen–Thawed Buffalo Spermatozoa by Using Cytofluorimetric Determinations

Flow cytometry is a useful tool that provides an accurate, objective and rapid evaluation of semen quality. The use of this technique could significantly improve the quality of buffalo semen samples used in artificial insemination. This study was carried out to evaluate, by flow cytometry, frozen–thawed buffalo spermatozoa quality parameters such as sperm viability by SYBR‐14/propidium iodide staining; mitochondrial function by JC‐1 potentiometric probe; sperm chromatin stability (SCSA) by acridine orange; and acrosome reaction (AR) by FITC‐PNA staining. Semen samples from five Italian Mediterranean buffalo bulls were used. Sperm viability was not different between bulls and ranged from 33.4% to 43.6%. A consistent rate (55.1 ± 10.8%) of sperm cells showed high mitochondrial membrane potential (Δψ^high), with no significant differences between subjects. Sperm chromatin structure assay differed significantly between the five buffalo bulls; moreover, data showed high stability within each buffalo. DNA fragmentation indexes (DFI), such as %‐DFI, ‐DFI, SD‐DFI, were 11.2 ± 8.6, 153.3 ± 24.6 and 81.6 ± 21.2, respectively. Regarding AR, the percentage of acrosome‐reacted live (ARL) and acrosome‐reacted dead (ARD) spermatozoa was 0.3 ± 0.2 and 15.3 ± 5.5, respectively. This functional parameter differed significantly between buffalo bulls and showed high stability. Following to Ca²⁺ ionophore A23187 for 3 h, AR significantly differed between subjects and was characterized by an increase in both ARL (10.8%) and ARD population (22.0%). This study indicates that flow cytometry could be a useful tool for a quick multiparametric evaluation of sperm quality in buffalo. In particular, SCSA and AR resulted in sperm functional parameters sensitive enough for the diagnosis of frozen‐thawed semen fertilizing potential.     

Comparative analysis of in vitro proliferative,migratory and pro-angiogenic potentials of bovine fetal mesenchymal stem cells derived from bone marrow and adipose tissue

Veterinary Research Communications - Mesenchymal stem cells (MSCs) are found in virtually all tissues, where they self-renew and differentiate into multiple cell types. Cumulative data indicate...