Immunolocalization of steroidogenic enzymes in the corpus luteum and placenta of the Japanese Shiba goat

In this study, we performed immunohistochemistry of cholesterol side-chain cleavage cytochrome P450 (P450scc), 3beta-hydroxysteroid dehydrogenase (3betaHSD), cytochrome 17alpha-hydroxylase P450 (P450c17), and cytochrome P450 aromatase (P450arom) in the corpus luteum and placenta of Shiba goats. The aim was to clarify the steroidogenic capability of the corpus luteum and placenta of Shiba goats. Ovaries containing corpora lutea were obtained from four adult Shiba goats during the luteal phase (day10; n=2) and pregnancy (90 and 120 days of gestation). Placenta was obtained from one Shiba goat on day 120 of gestation. The sections of the ovaries and placentae were immunostained using the avidin-biotin-peroxidase complex method (ABC) with polyclonal antibodies generated against steroidogenic enzymes of mammalian origin. All luteal cells expressed P450scc, 3betaHSD, P450c17 and P450arom. The distribution of P450scc, 3betaHSD, P450c17 and P450arom were not different during the luteal phase and pregnancy. P450arom showed a weak positive staining in late pregnancy (120 days). In addition, immunoreactions for P450c17 and P450arom were observed in syncytiotrophoblast of the placenta of one Shiba goat. These results indicate that, in Shiba goats, corpus luteum is not only an important source of progesterone but also has the ability to synthesize androgen and estrogen during the luteal phase and pregnancy. Also the placenta has the ability to synthesize androgen and estrogen in late pregnancy.     

Genetic polymorphism of plastid DNA in Tunisian date-palm germplasm (Phoenix dactylifera L.) detected with PCR-RFLP

Fifteen plastid fragments were amplified from a set of Tunisian date-palm accessions by PCR with consensus primers and analysed by RFLP. Polymorphic DNA bands were obtained as reliable molecular markers to estimate genetic distances among the accessions and to examine their phylogenetic relationships. The ctDNA PCR-RFLP method permitted the identification of two haplotypes that differ in the presence or absence of the HinfI restriction site. Phenetic groups composed of cultivars clustered together but does not constitute monophyletic groups. This typology agrees with the haplotypes' organization and provides a common genetic background within the implied varieties.     

Aloe vera long-term saline irrigation increases contents of hydrogen peroxide,lipid peroxidation and phenolic compounds

Salt stress is more and more becoming a serious problem in the world especially if we consider its damaging effect on the plant growth and yield. The cultivation of medicinal plants, such as Aloe vera, might be an alternative for the saline water use and salt-affected soils occupation. Aloe vera, commonly known as aloe, is one of the primary medicinal plants with multipurpose applications going from pharmaceutical to cosmetic aspects with a promising economic return. Aloe plants were cultivated and irrigated, for 14 months, with drinking water (C0) and with two levels of salt (C1 and C2). Changes in growth, hydrogen peroxide (H₂O₂), lipid peroxidation and phenolic compounds were examined in leaves at harvest. Depressive effects of salt irrigation on the plant growth parameters and a perturbation in inorganic ion contents were found especially with a high level of salt in the irrigation water. The intracellular oxidative stress was evaluated with the H₂O₂ production. Our results showed that the H₂O₂ content increased with the accumulation of the toxic ion (Na) in the leaf tissues. In addition, lipid peroxidation, measured by the malondialdehyde (MDA) level, increased as well with salt augmentation in the irrigation water. In response to salt stress, Aloe leaves showed a significant increase in the levels of phenolic compounds too. These results suggest that Aloe can be planted in soils affected by salinity and irrigated with salt water at least at a moderate concentration used in the present study.     

DNA extraction from bovine mummified fetuses and detection of factor XI gene deficiency in the mummies

Genomic DNA extracted from bovine mummified tissue is valuable material for detection of some genes that may contribute to fetal abnormalities. In this study bovine genomic DNA was extracted from the hardened tissue samples of ten bovine mummified fetuses. The amount of genomic DNA extracted from 2 g of the mummified tissues by the phenol/chloroform-ethanol method was low (less than 4 microg/ml) for all samples. The extracted DNA was then amplified by the GenomiPhi DNA amplification system. After amplification, the amount of DNA was increased to more than 100 microg/ml for all samples. This amplification system was shown to be a good tool for amplifying the genomic DNA of the mummified fetuses. The amplified genomic DNA was used for testing the mummies for Factor XI gene deficiency, an autosomal recessive deficiency involved in the early stages of the intrinsic blood coagulation pathway. Exon 12 of the Factor XI gene of the mummies was amplified by PCR. Two of the ten mummified fetuses were heterozygous for the Factor XI gene as indicated by the presence of two amplified DNA fragments of 320 bp and 244 bp. Factor XI deficiency has already been described in Holstein cattle. However, no report is available for bovine fetus. In this study, DNA was extracted and amplified from the bovine mummified fetuses, and the samples were successfully tested for Factor XI gene deficiency in the mummies.     

Factor XI mutation in a Holstein cow with repeat breeding in Japan

Factor XI deficiency is an autosomal recessive coagulopathy in Holstein cattle. Affected cows have a tendency to show repeat breeding. Forty repeat breeding Holstein Friesian cows were selected and tested for the Factor XI mutation. Genomic DNA was isolated from the blood of the cows (n=40). Exon 12 of the Factor XI gene of the cows was amplified by PCR. One repeat breeding cow was heterozygous to the Factor XI mutation as indicated by the presence of two DNA fragments of 320 bp and 244 bp. The insertion of the 76 bp in the heterozygous cow was confirmed by DNA sequencing. The heterozygous cow was in her fourth lactation. She gave birth to male twins at the last calving. She was inseminated artificially four times after the last calving. Factor XI deficiency in cattle has been reported in different countries. However, no case was reported in Japan. This might be the first to report Factor XI mutation in Holstein cattle in Japan.     

The bleaching induced by the phenylpyridazinone herbicide Metflurazon in Chlorella is a metabolic and reversible process

The treatment of synchronized cells of the green alga Chlorella fusca under photoautotrophic conditions with metflurazon (SAN H 6706; 4-chloro-5-dimethylamino-2-(α,α,α-trifluoro-m-tolyl-3(2H)-pyridazinone)) induces a bleaching process and results in white-appearing cells. This process of bleaching was followed by quantitative analysis of cell growth and cell division, photosynthetic oxygen evolution, respiratory oxygen consumption, and of pigment pattern at 0, 6, 12, 18, 24, 48, and 72 hr after incubation with different concentrations of metflurazon. Increasing concentrations of metflurazon gradually affected cell growth of Chlorella measured as increase in cell diameter. Cell division was inhibited completely with 1, 10, and 100 μM metflurazon. Photosynthetic oxygen evolution and respiratory oxygen consumption were not inhibited by 1 μM metflurazon during the first 6 hr; after this time a gradually increased inhibition was observed. Both parameters were inhibited by 100 μM metflurazon immediately after herbicide addition. A detailed analysis of the pigment content during the bleaching process revealed that: (a) The bleaching of Chlorella cells by metflurazon is not a simple photochemical process like the photobleaching of boiled cells, but is directed by the active metabolism of Chlorella itself. (b) The bleaching process is characterized by two phases: an accumulation of pigments followed by their degradation. The accumulation phase extends to 6 hr after herbicide addition. (c) During the accumulation phase, chlorophyll is accumulated to 380 and 106% in cells treated with 1 and 100 μM metflurazon, respectively, compared to the initial pigment content. The breakdown of chlorophyll, however, during the degradation phase is 5 times faster in the 1 μM treatment than in the 100 μM treatment. This difference resulted in the faster appearance of white cells with the low metflurazon concentration. (d) During the accumulation phase in the 1 μM treatment, the biosynthesis of chlorophylls, xanthophylls, and carotenes is inhibited by 56, 74, and 78%, respectively, when compared to a nontreated control. When related to the initial amounts, chlorophylls, xanthophylls, and carotenes are accumulated to 380, 230, and 153%, respectively. However, the synthesis of violaxanthin is specifically inhibited, followed by α-carotene. During the degradation phase, violaxanthin and α-carotene again, are the most rapidly disappearing pigments. Continuous culturing of white Chlorella cells resulted in a regeneration to green cells after 96, 240, 384 hr for 1, 10, and 100 μM metflurazone, respectively. The bleaching of Chlorella by metflurazon is evidently dependent on a functioning metabolism and is itself a regulated disassembly of the photosynthetic apparatus, which is reversible and not lethal.     

Virulence traits of avian pathogenic (APEC) and fecal (AFEC) <Emphasis Type="Italic">E. coli</Emphasis> isolated from broiler chickens in Algeria

Avian pathogenic E. coli (APEC) is the etiologic agent of avian colibacillosis, the most common disease responsible for chicken morbidity in the world. Although multiple virulence-associated factors were identified, their prevalence in Algeria is still poorly known. In the present research, 92 avian pathogenic E. coli (APEC) isolates were recovered from broilers with clinical signs and lesions of colibacillosis. In addition, 32 E. coli isolates collected from feces of healthy birds (AFEC) were included for comparison. All isolates were investigated by PCR for the presence of a total of 11 virulence-associated genes described for avian pathogenic (iroN, ompT, hlyF, iss, iutA, and fimC) and diarrheagenic E. coli (eae, stx, elt/est, ipaH, and aggR). The sensitivity of 39 APEC isolates to 16 antibiotics was also determined using antimicrobial pretreated microplates. Here, we report that 98% of the examined isolates host at least one of the tested virulence factors. The most prevalent genes in APEC were iutA (90.6%), ompT (86.9%), and iss (85.8%); whereas, iutA (78.1%), fimC (78.1%), and iroN (68.7%) were the highest prevalent genes in AFEC. Our data showed that none of the AFEC isolates harbor any of the tested diarrheagenic genes. Moreover, only elt/est (5.4%), stx (2.1%), and ipaH (2.1%) genes were carried by APEC isolates. We further established that ceftazodime, ceftiofur, mequindox, amoxicillin/clavulanic acid, and meropenem were the most efficient antibiotics against the analyzed APEC isolates. Overall, our findings provide more insights about APEC and AFEC virulence potential in Algeria which could participate in the fight against colibacillosis.     

Studies on Normal Haematological and Serum Biochemical Values of the ‘Hijin’ Racing Camels (Camelus dromedarius) in Kuwait

Haematological estimations and serum biochemical analyses were made on 100 samples collected from clinically healthy Hijin racing camels (Camelus dromedarius) in Kuwait. The red blood cell counts, packed cell volume, haemoglobin concentration, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration and total white blood cell counts were estimated. In the serum biochemical analyses, sodium, potassium, iron, calcium, phosphorus, magnesium, total bilirubin, blood urea nitrogen, creatinine, glucose, total protein and cholesterol concentrations were measured, as were the alanine aminotransferase, aspartate aminotransferase, -glutamyltransferase, creatine kinase and alkaline phosphatase activities. The results are discussed in relation to other findings reported in camels.     

Percutaneous ultrasound-guided over-the-wire catheterization of the portal and hepatic vessels in cattle

A new method for catheterization of the portal and hepatic veins in cattle by means of the over-the-wire system was investigated to maintain more reliable long-term patency of catheters. Four cattle were used to evaluate the success rate, patency and safety of the procedure. The catheters, coated by urokinase were patent as long as they were in situ. In addition, the introducer was useful to prevent the catheter from being broken. No complications developed during the10 days after the procedure. Two cows were then euthanized. Post mortem findings were minimal. The results of the study reported here are promising, the benefits are significant and there is no apparent disadvantage to its use.     

Characterization of the Exoglucanase-Encoding Gene PaEXG2 and Study of Its Role in the Biocontrol Activity of Pichia anomala Strain K

ABSTRACT The PaEXG2 gene, encoding an exo-beta-1,3-glucanase, was isolated from the biocontrol agent Pichia anomala strain K. PaEXG2 has the capacity for coding an acidic protein of 427 amino acids with a predicted molecular weight of 45.7 kDa, a calculated pI of 4.7, and one potential N-glycosylation site. PaEXG2 was disrupted by the insertion of the URA3 marker gene, encoding orotidine monophosphate decarboxylase in strain KU1, a uracil auxotroph derived from strain K. Strain KU1 showed inferior biocontrol activity and colonization of wounds on apples, compared to the prototrophic strain. Antagonism and colonization were recovered after the restoration of prototrophy by transformation with the URA3 gene. Integrative transformation was shown to be mostly ectopic in strain K descendants (only 4% of integration by homologous recombination). PaEXG2 disruption abolished all detectable extracellular exo-beta-1,3-glucanase activity in vitro and in situ but did not affect biocontrol of Botrytis cinerea on wounded apples.